單分子免疫陣列技術在傳染病診斷中的應用
單分子免疫陣列（single molecule ar放雨ray，Simoa）技術，又稱“數字ELISA”，是近年來新開(kā林書i)發(fā)的一種(zhǒng)數字免疫分析方法，將(爸通jiāng)單分子計數用于蛋白質生物标記物的檢測。該方法可測量各種(zh雪亮ǒng)不同基質（血清、血漿、腦脊液、尿液、細胞提取物等）中的蛋白了姐質，檢測限可達飛摩爾（fg/mL呢舞）數量級，靈敏度較傳統ELISA提高約1 000倍。目前，該技術在一些重靜西大傳染病，如新冠肺炎、結核病、艾滋病、朊病山民毒病診斷中的優勢逐漸凸顯。Simoa技術可幫助監測新冠病毒感染進(jìn化紅)程，有望實現早期高靈敏度診斷，農城以區分新冠肺炎輕症和重症患者；有望幫助診斷活動鄉媽性結核病，并可以監測結核病治療效果；可討身更準确檢測人免疫缺陷病毒（HIV）免疫相關蛋白含量變化，提高這低早期診斷的靈敏性；可在羊瘙癢病臨床症狀出現務飛前檢測活體動物血清，提高癢病早期檢測能(nén暗數g)力，從而有望用于人類克雅氏症診斷。未來，通過(guò)開(kāi友爸)發(fā)相關病種(zhǒng)的檢測試劑盒，有望利用Simoa技空的術提高疫病的綜合診斷能(néng)力，還(hái)可進(jìn)錯快行疫苗免疫效果評價，爲疫病防控提供依據。Ap線區plication of Single Molecule 窗讀Array in Diagnosis of Infectio南美us DiseasesSingle molec美草ule array（Simoa理聽），also known as“digit請去al ELISA”，was a new digi離如tal immunoassay technolog刀如y developed in rece長快nt years，which was generally used外動 to detect prote舊跳in biomarkers via single molecule coun得遠ting. The technology could be applied 志醫to measure prot靜海eins in various specim子年ens，such as serum，plasma光嗎，cerebrospinal fluid，urine，cell 妹新extracts，etc.，with a limit水樹 of detection as low道算 as femtomolar（人店fg/mL），and an approx得現imately 1000-fold sensitivity c土道ompared to traditional ELISA. C雜計urrently，the techno司少logy was promisingly a樹城pplied in diagnosis of some maj音地or infectious diseases including COVID風她-19，tuberculosis鐘好，AIDS and prion virus. As the p來他rocess of COVID-19 infection could be 黑愛monitored with the help of the tec吃就hnology，it was expect高還ed that，by the technology，su空和ch infection could be early d化技iagnosed with high sensitivity so as民鄉 to distinguish patients with seve信暗re or mild symptoms，so w路光as the active tuberculosi雜空s，and relevant therapeutic ef行懂fect could be monitored；an坐間y changes of relevant immunog畫視enic protein content related to 去身human immunodeficiency virus（HIV）子媽could be accurately影們 detected to impro生遠ve the sensitivity of early di線門agnosis；serums of live anima低請ls could be detected p輛謝rior to any clinical是動 symptoms of sc畫看rapie to improve early detection capa長來city and then it吧坐 was expected to be use學遠d in the diagnosis of human Creutzf紙訊eldt Jakob disease in the 現錢future. It was also e鐵化xpected that，through develo城草pment of kits for corre船醫sponding diseases，the玩要 technology could b女站e used to improve comprehens很他ive diagnosis capacity，and evaluate t聽知he effect of vaccination to provide a站少 basis for diseas體紙e prevention and control.畫多全文下載鏈接：https://kns.cn東術ki.net/kcms/detail/detail.aspx?db白這code=CJFD&dbname=C民購JFDAUTO&filename=ZGDW202107017&麗一v=2vpJqQNi66HYIULKef58WwZV放花JVLDZg6qPdhbwq1MqCrryBSve7KdHzDvQhal雪花Vaqt
2021-07-26
新型免疫佐劑碳納米管的研究進(jìn)展
碳納米管具有多種(zhǒng)良好(hǎo)的生物學(xué)特性，是近嗎吃幾年科研工作者研究的熱點納米材料之一，可增強機體樹線免疫應答，提高特異性抗體水平，在免疫佐劑研發(錢靜fā)領域展現了良好(hǎo)的應用前景。碳納米管作爲免疫佐劑的優勢在于其但下有極高的比表面(miàn)積，可以非特異性吸報日附多種(zhǒng)藥物、基因、蛋白質等物質；可進(jìn)入免疫細胞熱能，且不破壞細胞膜結構，從而爲抗上下原運輸和遞送提供了有利條件。目前關于碳納米管作用機體免疫問湖應答的研究取得了一定進(jìn)展，但其增強機體免疫應答的确切作用機制尚未票文揭示，今後(hòu)需對(duì)碳納米管最佳的功能(néng物議)化修飾方式、最适的免疫劑量及免疫毒性三方面(熱日miàn)進(jìn)行深入研究。本文總結分析了碳納米熱睡管作爲佐劑的優勢及其應用研究情況，以及存在的潛在問生金題及下一步研究重點，以期爲碳納米管免疫佐劑的研發(fā)提供新思路。Re費在search Progress on海匠 the Carbon Nanotube as a Novel Immun學讀e AdjuvantWith excelle紙好nt biological propert做錯ies，carbon nanotu司聽be（CNT）has become on你道e of the key nano materials studied by件小 science researchers in rec了男ent years，which could improve immun歌亮e response and in如紙crease the level of specific antibo學離dies，with a good a日木pplication prosp愛土ect in the development of市謝 immune adjuvant. With e月人xtremely high specific surf熱又ace area，CNT，as 些但an immune adjuvant，could non-spec得要ifically absorb various drugs，genes，科學proteins and other 雨少substances，and enter 習近immune cells without damaging an請對y structure of cell mem草答brane for the purpose of providing訊你 favorable conditions to trans綠她port and delive議拿r antigens. At p睡生resent，the studies on the 志線effect of CNT on i照通mmune response have taken微歌 a step forward，but the得但 exact mechanism for enhancing immune醫醫 response has not been ident家匠ified. Therefore，the optimal 煙農functionalization，i為鐘mmune dose and immunotoxi也書city of carbon nanotubes should be f店南urther studied in the future. I員公n the paper，the放新 advantages and applicati亮師on of CNT as an adjuvant，as 大火well as possible 朋外problems and future research 紅人focus were analyzed內女 with a view to providing some new 木雪ideas for next step.全文下載鏈接：https://kn外個s.cnki.net/kcms/det畫村ail/detail.aspx?dbcode=CJFD&dbname=CJF樂森DAUTO&filename=ZGDW202件哥107016&v=2vpJqQNi6的司6F%25mmd2BSI2TTQzug1SfI0uAyhOwe2b3xgJ8有用u7TxG1doR7XzGeDmyRHrp5子問Vo
2021-07-26
雪山草雞感染傳染性法氏囊病病毒新型變異株的鑒定
傳染性法氏囊病病毒（infectious bursal disea電去se virus，IBDV）新型變異株正在我國(guó)廣泛蔓延，給商品肉雞見上群和蛋雞群帶來了新的威脅。本研多樂究對(duì)一個疑似發(fā)生非典型傳染性法氏囊病的地方又綠品種(zhǒng)雪山草雞群進(jìn)行了RT我懂-PCR檢測，并對(duì)分離的3株毒株進(jìn科說)行了基因測序及序列分析。結果顯示，引起(q飛工ǐ)該雞群發(fā)病的病原是IBDV新型變異株，屬票都于A2dB1基因型；這(zhè)3下生個毒株的VP2高變區編碼蛋白上含有IBDV變異綠南株的特征性氨基酸，也具有IBDV新型變異株的獨特氨基酸。結果提示，我國(g也小uó)地方品種(zhǒng)雞群中也有IBDV新型變異株的流行，現有的部分商品請短化IBDV疫苗已不能(néng)有效預防IBDV新型變坐村異株的流行。Identific外對ation of Novel Varia秒公nt Strain of Infectious Bursal Dis林東ease Virus in Xueshan讀森 ChickenNovel variant strains of infec能文tious bursal di通請sease virus（IBDV）have been sp多拍reading widely 火聽in China，posing a會廠 new threat to commerci錢房al broilers and場可 layers. In the study，a l聽綠ocal variety of Xueshan chicken with s土窗uspected atypica水件l IBD was detected by RT-PC在黃R，gene sequencing報家 and sequence analysis were then condu船也cted for the three isolat船行es. The results showed tha暗離t the disease was caused by 又飛novel variant strain of IBDV that b去音elongs to A2dB1我到 genotype；in addition to the chara慢些cteristic amino acids of variant員坐 strain，the unique amino ac木明ids of novel variant IBDV were als明可o contained in the hypervaria子湖ble region of VP2 enco日拿ding proteins of the three isol快子ates. It was indicated th吃跳at the novel variant IBDV could not be 商行effectively contr我大olled by some current commer呢對cial vaccines，as it has been spread體朋ing in local chicken in Chi物短na.全文下載鏈接：https://答請kns.cnki.net/kcms/detail/detail技南.aspx?dbcode=CJFD&dbname=CJFDAUTO&filen兵雜ame=ZGDW202105022媽自&v=2vpJqQNi66EG9nURkPtQqnsBL45事舊kjWFuSxjcE%25mmd2BxMFYTC%25mmd2FNabg視藍exGz2EDjIZ%25mmd2BVgWY
2021-05-17
非洲豬瘟病毒抗體間接ELISA檢測方法的建立與應用
爲建立一種(zhǒng)檢測非洲豬瘟病毒（ASFV）抗體的間接ELIS上問A（iELISA）方法，對(duì)構建的ASFV P30我資基因表達工程菌誘導表達後(hòu)，將(jiāng)獲取的重組ASF個紅V P30蛋白進(jìn)行純化和West哥行ern-blot檢測，然後(hòu)以純化的重組蛋白爲抗原，建慢火立了ASFV抗體iELISA檢測方法，并進(jìn)行了特關文異性、靈敏性、重複性試驗；同時(shí)與基于ASFV P30-2Hi做請s6蛋白（N、C末端各融合1個Hi自話s6标簽）的iELISA方法進(jìn)行獸醫臨床樣(yàng)本比較試驗錯音。結果顯示：重組ASFV P30蛋白和窗車重組ASFV P30-2His6蛋白在Western-blot檢測中，均老南能(néng)與豬ASFV陽性血清産算妹生特異性雜交帶；基于重組ASFV P3站器0蛋白iELISA的最佳反應條件爲，抗原蛋白包被(b些謝èi)質量濃度20 μg/mL、血清樣(yàn兒好g)品稀釋度1:1 000、酶标二抗稀釋度1:40 000、血清樣(yà不放ng)品檢測OD450陽性結果臨界值0.22。該方法僅對(duì)ASFV陽聽了性血清呈特異性反應，1:3 200稀釋的陽性血遠喝清仍可檢出，批内試驗和批間試驗變異系數均小和生于10%，可以消除His标簽所造成(chéng)的假陽性話數反應。本研究建立的ASFV P30 iELISA檢測方法爲ASFV抗體檢測冷懂提供了一種(zhǒng)有效手段。Establishment and去小 Application of the iELISA for D妹習etection of Antibod去會ies against ASFVIn order to establi是飛sh an indirect enzyme河些-linked immunosorbent assay（iELISA）for著場 detecting antibodies 問近against African swine f民明ever virus（ASFV），the gene express店裡ion engineering st開生rain of recombina輛快nt ASFV P30 was induced離日 and expressed，followed by p厭頻urification and Western-blot 工電detection of the recombinant proteins門喝，then the iELISA method was establishe鐘坐d by taking the pur樂金ified recombinant protei藍數ns as antigens，its s區答pecificity，sensitivity and repea唱匠tability were subsequently eval自哥uated. Meanwhile，it was花歌 compared with the i錯煙ELISA based on ASFV P30-2His6 prot日刀ein（one His6 tag was fused 畫內respectively at N and C terminals）能吧for veterinary clinical samples. T費對he results showed that，by Wes北制tern-blot detection，both the reco很不mbinant proteins of A銀煙SFV P30 and ASFV P30-2His6 懂鄉could produce specific hybrid band唱光 with ASFV posit麗分ive serum；the optimal reaction conditi湖用ons of iELISA based on the recombina森謝nt ASFV P30 proteins were 20 μg/m電得L mass concentration of antigen prot錢場ein coating，1:1 000 dilution of serum 吃醫samples，1:40 000 dilution of goat a雜志nti-swine IgG conjugate，and 0.22 快爸critical value of positive serum sa吃高mples OD450. Th跳林e established method was specifi黑光c only to ASFV positive serum，1:話大3 200 diluted posit分員ive serum could still be detected紅離，and the coefficients of variation（C筆東V）of intra assay熱機 and inter assay wer理從e both less than廠店 10%，which could eliminate any fa說不lse positive reaction caused by His音個 tag. Therefore，an eff讀有ective tool was provided for d照一etecting antibodie要很s against ASFV b喝議y the establishe又市d ASFV P30 iELISA in the s但就tudy.全文下載鏈接：https:海們//kns.cnki.net/kc服湖ms/detail/detail.aspx?dbcode=C可公JFD&dbname=CJFDAUTO&filename=ZGDW2021吧上05021&v=2vpJqQNi66FSzfXYeD5Lvm3vSgcyl2市技HyKfSv6OXd%25mmd2BKgpa日的t1OHHXYyFn4oslWpcnF
2021-05-17
五種(zhǒng)重要犬病毒微流控芯片檢測方法的建立及應用
狂犬病毒、僞狂犬病毒、犬瘟熱病毒、犬細小病毒和犬冠狀病毒是引起(q廠玩ǐ)犬科動物重要傳染病的5種(zhǒng)病毒，嚴重威脅著(zhe)犬科動物的煙兒健康，是出入境犬科動物重點篩查的緻病因子。爲提升口岸進(jìn)出境中懂犬科動物檢疫工作效率，建立了上述5種(zhǒng)病毒為熱微流控芯片檢測方法。在建立5種(zhǒng)病機什毒環等溫擴增檢測技術（loop media鄉紙ted isothermal amplifica愛朋tion，LAMP）的基礎上，將(jiāng)這下雪(zhè)些LAMP檢測體系固化在同一塊塑料芯又少片上，制備了同步高通量快速檢測微流控芯片，并將(ji河會āng)其應用于臨床檢測。結果顯示：微流控芯片檢測方法暗技具有良好(hǎo)的特異性，含有目的基因的陽性樣(yàng)本僅在日服芯片上相應病毒反應槽出現顯著擴增，而其他病要靜毒反應槽未出現擴增，彼此無交叉反應；微流控芯片的敏感性與LAMP方法一緻；短冷通過(guò)檢測不同時(shí)期收集的感染犬瘟熱病毒、犬細小病毒和犬冠狀森紅病毒臨床樣(yàng)本，證實建立的微流控芯片檢測方法具有良好(hǎo)的穩定科見性和可靠性。與現有核酸檢測方法相比，微流控芯志校片可以在受試核酸上樣(yàng)40 min内完成(chéng)上述5雜習種(zhǒng)犬病毒的快速篩查，從而滿足進(jìn市河)出境犬科動物快速檢疫的需求。Es科這tablishment and Applicatio是討n of the Microflui子微dic Chip Assay for Five Kinds of 來通Major Canine Viruses Rabies v自照irus，pseudorabies virus，c那訊anine distemper virus，canine parvovi頻放rus and canine coronavirus生腦 are five kinds of viruses that 業會cause important infectious 近錯diseases of canine，which s為紙eriously threate要美n the health of canine，and ar拍那e the key pathogenic f光什actors to be screened and 務自examined by the exit-ent黑資ry ports. In order請了 to improve the 藍從efficiency of port entry-exit quarant匠我ine for canine，a microfluidic很相 chip assay for the above five匠什 viruses was establishe土書d. Five loop-mediate明電d isothermal amplification（L房紙AMP）assays were specifically es了家tablished and then solidif白一ied on the same plastic ch睡日ip to prepare a microfluidic ch時下ip with high-throughput森件 and rapid detecti要南on capacity for clinic作紅al detection. The resu技又lts showed that the established assay h友不ad a good specificity where pos厭西itive samples with 快快targeted genes were obviousl山自y amplified only in t內技he reaction tank 有如in corresponding viruses instead o船討f other viruses，an子的d no cross reaction with each o友事ther was found；the sensi問飛tivity of microfluidic 少大chip was consistent with that票弟 of LAMP；the establ高行ished assay was wit民還h good stability and reliabili個坐ty as verified through detectin的家g clinical samples infected with ca行但nine distemper virus an男好d canine parvovirus and collected i讀月n different perio愛紙ds. Compared to existing nucleic aci工事d detection methods，the microfluid下作ic chip could be 間鐘used to complete rapid screening of城他 the above five viru科區ses within 40 minutes after 媽舞the loading of nucleic acids，which c資物ould satisfy the needs of rapid裡在 quarantine for entry-exit canine ani愛道mals. 全文下載鏈接：https購來://kns.cnki.net/kcms/detail/detail遠服.aspx?dbcode=CJFD&算很dbname=CJFDAUTO&filename=房亮ZGDW202105020&v=2vpJqQNi66EdimcaTwAcYhu綠相aBu2kchUvfLZovQccWmMOE7GMIQI0zM暗務9H4K4ihNQs
2021-05-17
禽白血病病毒抗原高敏熒光微球快速定量檢測方法的建立
爲建立快速定量檢測家禽樣(yàng)本中禽白血為討病病毒抗原的方法，對(duì)禽白血病病毒p27蛋白單克隆抗體細胞株2E線北5和3D5進(jìn)行複蘇、鑒定，通過(guò)反應條件優化，建立了笑相禽白血病病毒抗原高敏熒光微球快速檢測方法。的下該方法能(néng)夠快速定量檢測家場但禽樣(yàng)本中禽白血病病毒，敏感性高、特異性資都強，對(duì)其他家禽病原無交叉反應且具有良好(hǎ有他o)的檢測重複性。對(duì)采自全國(guó)的240份臨床家一湖禽樣(yàng)品進(jìn)行檢測，同時(shí)用美國(g的呢uó)IDEXX公司生産的EL鐘冷ISA抗原試劑盒開(kāi)展比較，結果發(fā)現兩(liǎng)種(算答zhǒng)方法符合率達93.多民98%，其中陽性樣(yàng)品符合率爲90城的.83%，陰性樣(yàng)品符合率爲95.83%；組内與組間變異系數司影分别低于10%和15%。本研究建立的禽白血病病毒p27抗議唱原高敏熒光微球快速檢測方法，特異性高、敏感性強弟藍，操作簡單、快速，具有較高的應用推廣價值。E下筆stablishment of the High遠白ly Sensitive and Rapid Fluore就銀scence Quantitative Detection行數 for Avian Leukemia VirusIn歌訊 order to establish a rapid an錢麗d quantitative method 又影for detection of avian l音月eukosis virus（ALV）in poultry samples，2E技哥5 and 3D5，the monoc學作lonal antibody cell strains of ALV樹坐 p27 protein were revived and identif湖費ied，a highly sensitive and rap農習id fluorescence quantitative detection女小 method，with high sensit紅關ivity and specificity and go數妹od test repeatability and no cross rea刀黃ction to other pathogens鐘購，was established through o錯員ptimization of reaction conditions，by熱日 which，ALV could be什年 rapidly and quan分短titatively detected fro空章m poultry samples. 240 自筆clinical samples collec少近ted across China were detected and co愛如mpared with ELISA kit 請視produced by IDEXX. It物外 was found that the co山她incidence rate of th如風e two methods was 93.98%，in whic紅志h the coincidence rate of positive 河在samples was 90.83%，and that of ne通錯gative ones was 95.83%. The coefficien下湖ts of variation in intra g空地roup and inter group were lower t制金han 10% and 15%，re麗錯spectively. In conclusion，the es理爸tablished method窗也 was worthy of bein服熱g applied and expanded as su遠鄉pported by its high s就跳pecificity and sensitivity，easy and ra妹近pid operation. 全文下載鏈接：https:/行湖/kns.cnki.net/kcms/detail/d上鄉etail.aspx?dbcode=CJFD&dbname=分看CJFDAUTO&filename=ZGDW20了事2105019&v=2vpJqQNi66GeYlL7h4X%2友短5mmd2FfETw9jF5iXTqBJjpI7l9VOef8tuE2WJ鐘煙08X8bEbqL%25mmd2國費FnyF
2021-05-17
雞肝破裂血水綜合征病因初探
近幾年，在雞群中出現一種(zhǒng)以肝破裂血水爲主山議要特征的疾病，簡稱爲肝破裂血水綜合征，給養殖單位造成(笑報chéng)較大經(jīng)濟損失。爲探究雞肝破裂血水綜服民合征發(fā)生原因，采集11亮商省區市9種(zhǒng)品系雞的458小新份臨床肝破裂血水樣(yàng)本，開(kāi)展病原分離以及分子生物錢影學(xué)檢測、病理學(xué)診斷、内毒素檢測和動物高現試驗，并結合流行病學(xué)調的白查進(jìn)行病因探讨。病原分離結果顯示：檢測什來到大腸杆菌14份，占比2.8%；沙門的關氏菌7份，占比1.5%；未檢測到彎曲杆菌以及未分離到相關病毒。PCR、就看RT-PCR結果顯示：檢測到禽腺病毒（FA吧暗dV）3份，占比0.66%；禽戊用可型肝炎病毒（HEV）1份，占比0.22%；未檢測到雞傳染性貧血服歌病毒（CIAV）、馬立克病毒（MDV）、禽白血病病毒（ALV）。Rand街路om PCR檢測未發(fā)現RNA和DNA病毒；内火兒毒素檢測發(fā)現15份樣(yàng)本超标，占比3.3%。動物試驗結果顯示玩水，腹腔攻毒不能(néng)複制出臨床肝破裂血水表征。選取11省區人還市99份有代表性的肝髒樣(yàng)本，固朋秒定後(hòu)病理學(xué)診斷發也放(fā)現：無疑似髓樣(yàng)白血病/白血病農村病理變化；疑似細菌造成(chéng)的壞死性肝炎4份，占比4.04雪窗%；疑似禽網狀内皮細胞增生病并發(fā)血管瘤病理變化1份，占比2.廠現02%；澱粉樣(yàng)變94份，占話司比93.94%。流行病學(xué)調查發(fā)現：該的愛病在全國(guó)各地均有發(fā)病，發(fā)線習病雞品種(zhǒng)和日齡廣泛，無明顯季節性；同村技一雞場不同品種(zhǒng)，在飼料、免疫和管理都(dōu)一緻的情煙空況下，有的品種(zhǒng)肝破裂血水，有的品種(也習zhǒng)正常。以上研究表明，近幾年雞群中出現的肝破裂血水綜合征病例與白嗎常見的可引起(qǐ)雞肝破裂出血的因素無明顯相關性，吃兵初步分析與雞體本身存在遺傳自身免疫性缺陷因素有關，機體反應過(guò外術)度，出現細胞因子風暴綜合征導緻雞肝髒澱粉樣(yàng)變出血，從而出現又男肝髒破裂血水的臨床症狀。Exploration on the技和 Causes of Chicken Liver Hemor哥亮rhagic Syndrome笑美In recent years，an animal disease多務 mainly characterized 畫習by liver hemorrhag來她e（liver hemorrhagic sy他子ndrome）appeared in chicken，resul但北ting in huge economic loss to p畫購oultry industry. In ord話空er to identify relevant causes，麗友458 clinical lesion samples were 費舞collected from 9 chicken lines in 11 p訊時rovinces/distric請場ts/cities for pathogen isolation用要，molecular biological 謝習detection，pathological diagnosis，街間endotoxin test 輛哥and experiment on animals，從工the causes for the disease were di城月scussed based on epidemiological i匠南nvestigation results. It was shown tha雨微t，by pathogen isol亮刀ation，14 samples of Escherichia coli an校頻d 7 samples of Salmonella were detecte為地d out，accounting坐歌 for 2.8% and 1.5%，respectively，no 生門Campylobacter or other virus空知 was detected or isolated. It was conc看作luded that，by PCR and 微道RT-PCR，3 samples of fowl a行下denovirus（fadv）and on文光e sample of avian hepatiti購自s E virus（HEV）were detected out，accou信開nting for 0.66% and 0.2還年2%，respectively，no c雨體hicken infectious anemia virus（CIAV），森錯Marek's disease virus（MDV）and avian車麗 leukosis virus（ALV）were dete我著cted. No RNA or DNA virus吧內 was found by Random 分體PCR. 15 samples門子 exceeded the standa匠做rd for endotoxin，來物accounting for 3.3%. It was found 城校that，by experiment on animals，the sym下動ptoms of clinical liver hemor話妹rhage could not be reproduced by日店 intraperitoneal challenge. 99 represen內物tative liver samples were selec花自ted from 11 provinces/districts/鐘個cities，followed by being fix開生ed for pathological dia睡物gnosis，no suspected myelo自器id leukemia/leukemia p哥高athological changes were found，4現做 cases of necrotic hepat見資itis caused by suspe都聽cted bacteria，one suspected avian reti綠森culoendotheliosis complic鐘知ated with hemangioma and 94 cases 亮唱of amyloidosis were observed，accounting人裡 for 4.04%，2.02% and 93.94物算%，respectively. By epidemiological 麗到investigation，it was found tha要機t the disease occ去科urred all over t體文he country，covering 聽紙a wide range of varieties and age，witho明新ut obvious seasonality；for 樂雜different varieties in the same farm，so到跳me were with liver 金拍hemorrhage but others were 她少normal under consist物白ent conditions of feed，vaccinati綠又on and management.了黃 In short，the cases of liver 店音hemorrhagic syndrome found in rece師黑nt years were not related to the c亮通ommon factors leading to liver hemorr做但hagic，which was connecte唱靜d with the genetic a動間utoimmune defects in chicken a器木s preliminarily anal藍影yzed，specifically，th讀照e organism reacted ov嗎河erly，followed by cytokine storm，result東從ing in amyloidosis bleeding大很 in chicken liver，and thereby clinical笑妹 symptoms of liver hemorrhage商月.全文下載鏈接：https://kns樂討.cnki.net/kcms/detail/detai多鐵l.aspx?dbcode=CJFD&dbna農會me=CJFDAUTO&filename=ZGDW妹要202105018&v=2vpJqQN市微i66Gk%25mmd2FmO路妹b8Nn1bPYoKcWscH1oT57w4oQOpUdlny們子lBTbdSc3QPJgPCq9eb
2021-05-17
無乳鏈球菌對(duì)奶牛乳腺成(c報答héng)纖維細胞金屬蛋白酶及u錯外PA系統表達的影響
爲研究無乳鏈球菌（Streptoco志志ccus agalactiae，S. agalac長作tiae）作用奶牛乳腺成(chéng)纖維細胞（BMFBs）後(hòu海作)對(duì)MMPs/TIMPs與uPA系統（uPA/PAI-1）吃雜表達的影響，以進(jìn)一步秒數闡明MMPs/TIMPs與uPA系統在調控細胞外基質（ECM）代謝的聽綠作用，將(jiāng)107 cfu/mL熱滅活S.場靜 agalactiae菌液作用道低于BMFBs，在作用後(hòu)劇就不同時(shí)間點（6、12、24木從、48 h）通過(guò)RT-qPCR和請謝Western Blot檢測M聽腦MP-2、MMP-9、TIMP-1個跳、TIMP-2、uPA、PAI-1 mRN些服A轉錄水平和相應蛋白表達水平，并用明膠酶譜法檢測MMP到冷-2、MMP-9的酶活性。結果顯示：熱滅活S. agalactiae作用後(明票hòu)，MMP-9、TIMP-2、uPA、PAI-1 mRNA轉錄水鄉劇平均有不同程度下降趨勢，但MMP-2 mRNA水平上調朋說，TIMP-1 mRNA水平呈也厭先上升後(hòu)下降趨勢；明膠酶譜法視頻檢測MMP-2的酶活性較MMP-9明顯，二者均在匠我作用6 h、48 h活性最強；MM美民P-2和MMP-9蛋白表達水平與對(duì)照組相比主要呈先升高後(hòu做物)下降趨勢，但在作用後(hòu)期MMP-2蛋白表達又出現了回升趨勢；T件拿IMP-1蛋白水平與對(duì)照組相比主線光要呈先下降後(hòu)上升趨勢，在作用前期與MMP-9蛋白表達相反，TI南愛MP-2蛋白表達與對(duì)照組相比呈上升影音趨勢；uPA和PAI-1蛋白水平與對(書體duì)照組相比均呈先降低後(hòu)升高再海報降低趨勢，均在24 h達到峰值，随後(hòu)呈下降趨勢。研究表明，熱滅子業活S. agalactiae菌液可誘導BMFB城錢s中MMPs/TIMPs表達，影響uPA系統參與調控MMPs的表達及活性的妹。The Effect of Inactivated S區時. agalactiae on 知木Metalloproteinase an兵媽d uPA System Expre舞務ssion in BMFBsIn order to st信街udy the effect of 低務Streptococcus agalacti謝說ae（S. agalactiae）on bovine mammary fib服工roblasts（BMFBs）and subsequen鐘白tly on the expres秒很sion of MMPs/TIM森數Ps and uPA system（uP跳照A/PAI-1），and to further clarif下章y the role of MMPs/TIMPs and u白身PA system in regulating extracell訊民ular matrix（ECM）metabolism，107cfu他讀/mL heat-inactivated S. aga信照lactiae was acted o媽會n BMFBs，the levels o藍開f MMP-2，MMP-9，TIMP-1，TIMP-2，u媽子PA，PAI-1 mRNA transcription 銀拍and corresponding protein expressio照讀n were detected by RT-qPCR and Wester睡地n Blot at different time poin門街ts（6，12，24 and 48 h火小），then the enzymatic activities of MMP小舊-2 and MMP-9 were det哥湖ected by gelatin z空術ymography. The results showed t弟懂hat after the reaction of heat資會-inactivated S. agalactia知家e，the level of t雨東ranscription of MMP-9，TIMP-2，uPA and P車雨AI-1 mRNA trended to reduce to 多白different extent，except that of MMP-2 m得姐RNA，and that of TIMP-1 mR鐘都NA increased firstly and th玩森en decreased；the enzymatic activity of 司好MMP-2 was more obvious上民 than that of MMP要笑-9 as detected by gelatin zymography，bo土體th of which were st訊內rongest in 6 and 8 h after reaction；as 銀藍compared with the contr自文ol group，the level of protei來不n expression of MMP-2 and MMP-9 t行紅rended to increase prior to decr為短ease，but the pr一國otein expression of MM朋河P-2 rose again at later stage；as compa好畫red to the control grou放高p，the level of protein of TIMP-1 show家林ed a trend of decreasing prior t公紅o rising，which was 請答contrary to that of M厭問MP-9 at early stag拍化e，and TIMP-2 trended to increase；t亮線he levels of protein of uPA and PA區道I-1 both decreased firs白業t then increased，and decreas得花ed finally，reaching the peak at 2知什4 h after reaction. In conclusion，the暗笑 expression of MMPs/TIMPs 劇從in BMFBs could be induced著森 by heat-inactivated S. agalactiae，whic明多h could influence uPA system in regula還白ting the expression and act訊暗ivity of MMPs.全一謝文下載鏈接：https://kns.cnki.net/亮山kcms/detail/detail.aspx?dbcode=CJFD&d輛會bname=CJFDAUTO&filename=Z厭道GDW202103017&v=票什2vpJqQNi66HD98xaKRkUCe2%25mm時煙d2FvY%25mmd2BdoL技房qNpkBm9OA0N3AHjQbEicL7krHGw89vj4Hl少民
2021-05-17
新疆野生北山羊小反刍獸疫病毒分子特征
爲了解新疆野生北山羊小反刍獸疫病毒（peste des petits ru這去minants virus，PPRV）的煙資分子特征，根據GenBank公布的PPRV基因組序列設計并合成(c紅化héng)引物，采用RT-PCR方法和基因測序技術獲得病毒全基因序列，應用些山分子生物學(xué)分析軟件，對(duì)分離的P高輛PRV毒株進(jìn)行序列分析西爸。結果顯示，本次分離的PPRV毒株（China/XJAKS/2017）屬于明爸基因IV型，基因組全長(chán女山g)15 954 nt，編碼6種(zhǒng)結房朋構蛋白和2種(zhǒng)非結構蛋白；在系統進(jì行務n)化上，與國(guó)内新疆分離株China/XJBZ/2015家家、China/XJYL/2013同源率分别高達99.0%、99.6%腦都，與國(guó)外的巴基斯坦和塔吉克能文斯坦分離株親緣關系最近。結果表明，P訊鐵PRV已經(jīng)在家養動物與野生動物之間傳播。結果提示，PP年是RV傳入野生動物群爲我國(guó)消除PPR帶來極大困難，需加強我國(guó)紅微邊疆地區PPR免疫隔離帶建設，并對(好亮duì)野生動物PPRV分子流行病學(xué)特空算點進(jìn)行追蹤監測。 Molecular Characteri動來stics of PPRV I多數solated from Ibex in Xinjiang我路 In order to identify the 煙秒molecular characteristics of pes匠司te des petits ruminants virus（PP城國RV）in ibex in Xinjiang，primers we公書re designed and synthesized acco數暗rding to the gen請又ome sequence of PPR事區V published in GenBank，the whol時長e gene sequence of the v報劇irus was obtained by 友舊RT-PCR and gene sequencing，then the is綠錯olated PPRV strain was sequenced 問她and analyzed by molecular biology a文長nalysis software. The results showed t熱分hat the isolated PPRV strain（China/匠對XJAKS/2017）fell into genotype IV，with a員們 total genome length of 15 954白日 nt，encoding 6 struc商道tural proteins and風高 2 non-structural proteins是計；for phylogenetic evolution，its 年兒homology rates wit分西h Xinjiang isolates inclu筆雜ding China/XJBZ/2015 and China/X線行JYL/2013 were 9舞車9.0% and 99.6%，respectively，an低話d it was closest rel如地ative to Pakistan舊議 and Tajikistan isol火厭ates. It was co空朋ncluded that PPRV had been 少大spreading among domestic文廠 and wild animals，especially wild anima家動ls，which brought difficulties 慢銀to eradicate it in China，從關so it was necessary 小厭to strengthen the construction of isol民船ation zone with v下車accination against FMDV in border 冷房regions in China，and to track an到影d monitor the mo日時lecular epidemiological charac新兒teristics of PPRV in 姐白wild animals.全文下載鏈接：https://kns.cnki.ne舊小t/kcms/detail/detail.aspx?dbcode=CJF開電D&dbname=CJFDAUTO&filename=ZGDW202間花105008&v=2vpJqQNi66FB38爸那KGubt17K3%25mmd站雨2BS%25mmd2BOWJtAMbK55OsDUUt熱雪zg45nPuZOLL3I5PoszpeXv
2021-05-17
我國(guó)西部地區農牧民包蟲病防控認知情況及其影響因素
爲掌握新疆、青海、甘肅、四川和西藏等通工西部5省區農牧民包蟲病防控相關知識、态度和行爲（“知信行”），分析美議影響“知信行”水平的關鍵因素，對(duì)1 213名農牧民進(j服喝ìn)行問卷調查，用百分比描述“知信行”的每個條目，采用t檢驗或方差分析，比北綠較不同人群的“知信行”水平差異。結果顯不低示：西部5省區農牧民對(duì)包蟲病防控相關知識的知曉率爲57.舞老0%，其中感染途徑知曉率最低，爲制答30.8%；55.6%的農牧民對(術但duì)包蟲病防控持有積極态度，68.3%的農牧民對(duì站數)包蟲病防控有較好(hǎo)的行爲習慣。當銀光地農牧民獲取包蟲病防治知識的途徑多來自醫生和傳統宣傳資料，分别討章占60.7%和48.2%，未來還(hái)希東事望通過(guò)電視、廣播和網絡等大衆媒體獲取。青影員海省農牧民以及文化程度初中及以上、民族爲漢族的農牧民具有較高的認知做弟水平。結果表明，西部5省區農牧民對(duì)包蟲病的認知水平較低，獲取相關信息去子途徑較爲單一。建議加大宣傳力度，突出包蟲病源頭防控；利用當前主流新媒體，豐富多用宣傳方式，使宣傳内容、載體更加契合西部少個行數民族地區的生活和文化習慣，切實推進(jìn)農牧民防疫意得綠識和自我防護意識的提升。Awareness of Herdsmen 子城for the Prevention and Control of Ec用女hinococcosis and Potential Risk Fa那技ctors in Western ChinaIn order to inves通家tigate relevant knowledge，東又attitude and practice（KAP）o算音f herdsmen towards the prev術制ention and cont一從rol of echinococcosis in Western Chin家舞a including Xinjiang紙術，Qinghai，Gansu，Sichuan and Ti遠人bet，and to analyze the key f綠短actors for KAP level，a qu綠坐estionnaire investigation was 城民conducted for 1 213暗一 herdsmen，each item of 你北KAP was describ話能ed by percentage站路，and the KAP levels among d紅地ifferent popula吧器tions were compared 冷拍by t test or ANOVA. The商些 results showed that 廠司the average awareness又關 rate of herdsmen 東森for the knowledge of prevent都也ion and control of echinococcosis 綠鄉was 57.0%，and that fo相術r infection route was 30.8%，which wa嗎子s the lowest；55.6% of herdsmen were 自放active for the prevention of echinococ森輛cosis，and 68.3% performed bet區市ter behavior hab動快its. Local herdsmen could access to th在秒e above knowledge mostly from d喝快octors and traditional publicity船線 materials，account白低ing for 60.7% and 48.2%，respectiv物票ely，mass media such 唱什as television，rad內動io and Internet wer日車e also expected in the futu算黃re. The herdsmen wit林器h education back能愛ground of junior middle scho對能ol or above，and from Han ethnic 空年group in Qinghai Province were at雪空 high awareness level. It was concl畫中uded that the herdsmen knew little abo線雪ut echinococcosis術呢 due to single pathway to access to rel場什evant information. It was suggest工見ed to strengthen awareness ac訊器tivities with a priority of preventio男身n and control from source，enrich p友秒ublicity methods by use of current n小森ew media to make the contents and carri美樹ers conform to the living and cultura生算l habits in the ethni時黃c minority regions，and to effectively林又 improve herdsmen's awar雨線eness for prevention and control of th照們e disease and self-protection.歌這全文下載鏈接：https://kns.cnki.net/kcms/de音拍tail/detail.asp多間x?dbcode=CJFD&dbname中藍=CJFDAUTO&filename=ZGDW20210照作5001&v=2vpJqQNi66EDmTu0UXlUm6Q%25m業分md2BIvqB1ZwkysgLv0長離jd%25mmd2B2gd8fd民熱MS9VGzNWDhm8218%25mmd2B3
2021-05-17
腸衣傳統和新型腌制技術研究及應用進(jìn)展
我國(guó)是世界腸衣出口大國(guó)。爲生山使國(guó)内腌制腸衣企業全面(mi飛知àn)了解國(guó)際腸衣腌制工藝更新動态，及時(shí)跟蹤相關機構的科研為熱方向(xiàng)及研究進(jìn)展，本文綜行日述了傳統氯化鈉（NaCl）腌制腸衣技術和新型磷酸鹽補充幹鹽（P-salt）道媽腌制腸衣技術的研究背景、最新進(jìn)展及其應用範圍，介紹自數了國(guó)外更新腸衣加工工藝的上那操作模式。NaCl腌制技術一直是腸關廠衣防腐殺菌和常溫保存的主要方法，目前國(guó)際飛暗貿易中絕大多數半成(chéng)品和成(chéng)品腸衣采用該技術處理。N林通aCl腌制技術的殺菌能(néng)力與腌制腸衣中的鹽濃度即水活對年性、腌制溫度和pH等多種(zhǒng)因素聯系緊密。P-salt腌制技術火年能(néng)顯著提高腸衣的衛生狀況和機械長現特性，既可顯著提高腸衣灌注内容物時(shí)的潤滑度，又妹風能(néng)提高腸衣攜帶病毒的殺滅效果。一些相關研究成(區筆chéng)果已成(chéng)爲當前制定腸衣腌制問信處理工藝的重要依據。目前國(g錢拿uó)内外的腸衣腌制技術已由單一的店外NaCl腌制技術，逐漸進(jìn)我快入到傳統NaCl腌制和新型P-salt腌制共存的局光愛面(miàn)，并大有向(xià著話ng)後(hòu)者傾斜的趨勢。本文黃樹爲國(guó)内腸衣腌制技術的研究和儲備提供了參考。Research and紙校 Application Prog市唱ress of Tradition亮店al and New Salting Techniques fo那書r Casing for SausagesAs China has been 愛麗a major exporter of casing for saus但音ages in the world，the research bac輛北kground，latest prog黑拍ress and application scope of the sa讀她lting techniques by 男內traditional sodiu年湖m chloride（NaCl）and new phosph年鄉ate supplement dry salt（P-salt）were sum草章marized，and the operation mo北她de of latest casing proce區海ssing techniques in o拍光ther countries was introd務要uced in the paper，for the pur事也pose of learning update 海算trend of international salting te習們chniques，and tracing the resear道章ch direction and progres樂鐘s in relevant organizations. N事嗎aCl salting technology 短唱has always been 物冷the main method for antisepsis，工他sterilization and room temperatu老商re preservation of casings，a個文t present，most semi-fin門有ished and finished casings in玩化 international trade are treated with務黑 this technology. The bacter大雜icidal capacity of t子多he NaCl salting technology is clo近雨sely related to salt雜森 concentration（water妹音 activity，curing temperature，火北pH value and ot知就her factors）in sal有友ted casings. P-salt salting techniq長章ue can obviously improve sanitary cond紙月itions and mechanical properties of農快 casings，including b木睡oth the lubricity when cont頻還ents are filled into casin報場gs and the eradication算話 effect when viruses are carried by 銀愛casings. Relevant research results have自裡 been considered as the important道謝 basis for develo請服pment of salting technique for casings.去草 Currently，the salting technology 高藍of casings at h如看ome and abroad has gradually enter爸子ed into the coexistence of traditi照文onal NaCl and new p-美站salt salting technology，and ther對哥e is a great trend to the草車 latter. Some re車中ferences were thereby provi遠麗ded for the research and 用站reserve of casing salting te美子chnique in China.全文下載鏈接：https://kn化新s.cnki.net/kcms/detail/通答detail.aspx?dbcod照醫e=CJFD&dbname=CJFDAUTO&filename=ZGDW202新場105015&v=2vpJqQNi66EGh們市D5qw18P4wrvCIB9HA去樂3PPuI98VprWEkP2理年jNEHuAwXrt2M%25火動mmd2FhcJYgx
2021-05-17
液相色譜-串聯質譜法測定豬尿液中地西泮及其7這什種(zhǒng)代謝物殘留
爲建立快速測定豬尿中地西泮及其7種(zhǒng)代謝物殘留的超高效液相色譜-土慢串聯質譜（UPLC-MS/MS）确證方法區放，本研究優化了樣(yàng)品淨化的陽離子交換固相萃取柱（PCX柱）及其淋數輛洗洗脫條件。豬尿液酸化後(hòu)直接經(jīng)PCX柱淨化，依次用水睡到、60%甲醇水溶液淋洗，最後(hòu)用5%氨化甲醇洗脫；選用BEH 雨國C18色譜柱分離，UPLC-MS/校理MS進(jìn)行檢測，以基質匹配标準曲她會線定量。結果顯示：8種(zhǒng)藥物在0.3~20們草.0 μg/L範圍内具有良好(hǎo)的線性關系（R2≥工子0.995），檢出限和定量限分别爲0.1 μg個公/L和0.3 μg/L；各藥物在3個添加濃度下回收率爲73資機.6%~95.3%，日内、日間相對(duì資大)标準偏差分别爲2.9%~18.6%（n=6）和2在雨.2%~12.6%（n=3）。結果表明，本方法操作簡單、習月快速，靈敏度高、特異性好(hǎ音我o)。Determination of Diazepam 鄉老and Its Metabolit腦他e Residuesin Swine Urine by UPLC-動站MS/MSIn order to es年還tablish an ultra performance 計近liquid chromatography-tandem mass 輛路spectrometry（UPLC-MS/MS）to determi聽討ne diazepam and its 路站seven metabolite residues（temazepam，見影nimetazepam，nordiazepam，ox件在azepam，nitrazepam，7-aminonitra拍商zepam and 7-aminonimetazepam）in 吧理swine urine，a cation exch舞討ange solid phase extractio懂行n column（PCX column）and its co雜亮nditions of elutio短山n were optimized. Swine ur喝件ine was directly purified by問讀 PCX column after being acidized，washe時匠d with water and 60% methano務費l water solution in外些 turn，and then eluted with 5% ammoniate理自d methanol；then the pre-東會treated urine sa生司mple was separated by算錢 BEH C18 column，de老你tected by UPLC-MS/MS，and qu話多antified by matrix-matched extern子村al standard curves. The r站小esults showed that a good li購開near relationship（R2≥0.995）among the 大坐eight drugs was obtained wi通冷thin 0.3 to 20 μg/L，the limits of detec看秒tion and quantification were個報 0.1 and 0.3 μg/L respectively；t什生he mean recoveries vari跳城ed from 73.6% to 95.3% at放關 the three concentr作美ations，the relative standard deviations你問 of intra and inter d冷那ay were 2.9% to 18.6%（n=6）and 2.2% 飛遠to 12.6%（n=3），re學年spectively. It was con國中cluded that the proposed method was 業筆simple and rapid with high sensitivity草村 and specificity.全文下載鏈接：https://kns.子近cnki.net/kcms/detail/detail.照個aspx?dbcode=CJFD&dbna醫廠me=CJFDAUTO&filename=ZGDW202103022&v化刀=2vpJqQNi66FHwRwrKctQIdf4s明費HDyT80dN7Vep7TY5yE2n5Tg湖制lxoDELE5pR1HP%2和兵5mmd2FaL
2021-03-16
獸用疫苗中BVDV、PCV2和PPV污染答制三重熒光定量PCR檢測方法的建立
爲建立可以快速同時(shí)檢測獸用疫苗中牛病毒性腹瀉病毒（BVDV）、豬麗家圓環病毒2型（PCV2）和豬細小病毒（PPV）3種(zhǒng)病原我著的方法，通過(guò)研究比對(duì)BVDV 5'音朋;-UTR、PCV2 Rep以及PPV NS1基因序列，分别湖們設計合成(chéng)了3對(duì)引物和3條熒光探針，在優化林技反應條件和反應程序後(hòu)，建立線女了一種(zhǒng)可同時(shí)檢測BVDV、PCV2對文以及PPV的三重熒光定量PCR方法，并對門區(duì)其敏感性、特異性進(jìn)行了評估。結果顯示，建立的三重熒光定量吃下PCR方法靈敏度高，對(duì)3種(zhǒng)病原核酸的最低檢測限均爲1兵商0 copies/µL；特異性強，與其他相關病原（舊子豬瘟病毒、豬繁殖與呼吸綜合征病毒、僞狂犬病視理病毒、非洲豬瘟病毒）均無交叉反應。結果表明這能，本試驗建立的三重熒光定量PCR适于疫苗中外源病毒的檢測，可用于疫苗等生店身物制品質量把控。Establishment of花愛 a Triple Fluorescent Quant他謝itative PCR for Detect還去ing BVDV，PCV-2 and PPV in Veterin是相ary VaccinesIn order 機紙to establish a rapid 店他method for simultaneously detecting街訊 bovine viral di行雜arrhea virus（BVDV），porcine 農小circovirus type 2東厭（PCV2）and porcine parvovirus（PPV）in vet湖筆erinary vaccines，three pairs of prim懂風ers and three fluorescent熱開 probes were de見線signed and synthesized through comparin計跳g the sequences of BVDV 5'-UTR，白件PCV2 Rep and PPA NS1 genes，f子笑ollowed by the opti自鄉mization of reaction算媽 conditions and procedures，a triple f子都luorescent quantitative PCR was establi行到shed for simultaneous關劇ly detecting BVDV，PC山對V2 and PPV，then its s對拍ensitivity and specificity were 高爸evaluated. The results showed that the雜男 established method was with high 家內sensitivity and specificity，老讀its minimum detection limits for the th房話ree pathogen nucleic acids were 10街綠 copies/μL，and failed明山 to react with other relevant path知拿ogens，including classi畫村cal swine fever virus（CSFV），por了了cine reproductive and respiratory s土新yndrome virus（PRRSV），pseudorabies呢下 virus（PRV），African 玩西swine fever viru音書s（ASFV）. It was conclude朋離d that the metho她車d was applicable for detection of exog能河enous virus in vaccine吧數s，and could be u大好sed to control the quality 就生of vaccines and other biologi議員cal products. 全文下載鏈接：https://kns.cnki畫司.net/kcms/detail/detail.aspx?db現是code=CJFD&dbname=CJFDAUTO&f媽商ilename=ZGDW202103021&相少v=2vpJqQNi66F74B如來tupbXSqOF6E0q2970nQnOlj%25mmd2F98%25m玩日md2FrBzx%25mmd2BbBHjr31kasB71O7ZIJ
2021-03-16
BVDV、BRV和BCoV Taq動媽Man三重RT-qPCR檢測方法的建立
爲建立牛病毒性腹瀉病毒（BVDV）、牛輪狀病毒（BRV）和牛冠狀病毒（B機裡CoV）的快速檢測方法，根據GenBank中登錄的B很舊VDV 5'-UTR、BRV NSP5和BCoV N基因序列設計南暗特異性引物和探針，通過(guò)優化反應體南和系和條件，建立了同時(shí)檢測上述3種上月(zhǒng)病毒的TaqMan三重RT-qPCR方法。該頻船方法僅對(duì)BVDV 5'-UT北工R、BRV NSP5和BCoV N基因擴增呈陽性，而對(duì)牛傳染性鼻氣坐木管炎病毒、牛副流感病毒、魏氏梭菌（A裡時型、B型和D型）、多殺性巴氏杆菌（A型和B型）等犢牛腹瀉相關病原擴增均呈陰性；藍商最低檢出限爲10拷貝/μL；組内和組間變異系數均小于2%。利用本研究建立的Ta制雪qMan三重RT-qPCR方法費妹對(duì)29份臨床樣(yàng)品進(jìn)行檢測，獲得BVD自飛V、BRV、BCoV的4種(zhǒng)混合感染型，其中BVDV與農湖BCoV的混合感染率最高（27.6%）；與已有單服村重RT-qPCR方法比較，發(fā)現兩(liǎng)種(zhǒng)方街的法檢測BVDV、BRV和BCoV的符合率分别爲身討100%、96.5%、100%。結果表明麗明，本研究建立的TaqMan三重RT-qPCR方法具有但技特異性強、敏感性高、重複性好(h水煙ǎo)、可行性高等優點，可爲今後(hòu)道得BVDV、BRV和BCoV共感染引起(qǐ)的牛腹瀉性疾病的玩朋鑒别診斷和流行病學(xué)調查提供新技術手段。Estab舞離lishment of TaqMan Triple RT-房區qPCR for Detectin樂們g BVDV，BRV and B亮相CoVIn order to establish a ra聽錯pid method for detecting bo近現vine viral diarrh服慢ea virus（BVDV），bovine rotavir民答us（BRV）and bovine coronavirus（BCoV），話機the specific primers and probes南麗 were designed according to the seq歌慢uences of 5'-UTR，BRV N鐵去SP5 and BCoV N genes r國林egistered in GenBank，followed by the章錯 optimization of拍術 reaction systems and cond南紙itions，then the TaqMa個我n triple RT-qPCR was establishe秒能d to simultaneously detect the above 北很three viruses. By which，positiv花影e results only occurred in the am女有plification of BVDV 5'-UTR，BRV NSP西公5 and BCoV N genes，but negative友暗 for infectious bovine rhin謝地otracheitis virus（IBRV），bovine 線很parainfluenza virus（BPIV），Clos路這tridium welchii（type A，B and 師現D），Pasteurella multocida（type A去黃 and B）and other patho訊土gens related to 亮司calf diarrhea；the minimum detect舊紙ion limit was 10 copies/μ業科L；and the variation coefficients of又習 intra-and inter-group were l分身ess than 2%. 29 喝來clinical samples were de坐討tected by the established method，跳請the four types of co-infection 雨是of BVDV，BRV and BCoV were obtained，in雨中 which，the co-infection r就章ate of BVDV and BCoV was hi快路ghest（up to 27.6%）. The coincide分路nce rates of current single RT-qPCR西站 and the establis費拍hed method for BVDV，BRV and BCoV we做亮re 100%，96.5% and 100%，respectively.樂家 As a conclusion，廠厭the established method could be applied睡玩 in differential diagnosis and epidem子嗎iological investigation against 慢志bovine diarrhoeal diseases ca如能used by the co-inf長小ection with BVDV，BRV and BCoV in th生吃e future due to 門工its advantages of go開腦od specificity，sensitivity，repeatabilit多雨y and feasibility. 全文下載鏈接：https://照明kns.cnki.net/kcms/detai他家l/detail.aspx?dbcode=CJFD&dbname=CJFD妹通AUTO&filename=ZGDW2021哥個03020&v=2vpJqQN也火i66FgWN7t02lf8J慢日APYtfwV3kJao7hAGUuiVll0QCZG9Q10FUq5hMJe計城Vk8
2021-03-16
PCR結合斑點雜交技術檢測禽多瘤病毒方法和志的建立及應用
鹦鹉幼雛病是由禽類多瘤病毒（APV）引起(qǐ)海會的多種(zhǒng)鹦鹉雛鳥死亡的急性病毒性傳染病，嚴重危害鹦鹉養殖錯些業的健康發(fā)展。爲提高分子生物學(xué)方法答內檢測APV的敏感性和特異性，對(duì)APV基因片段進習員(jìn)行克隆和序列分析，設計合成(chéng)1對(duì)特異務媽性引物，以VP1基因爲模闆，經(jīng)PCR擴增獲得731 bp的核苷路冷酸DNA，并用DIG标記，制備用于檢志身測APV的特異性核酸探針；對(du笑員ì)制備的探針進(jìn)行靈敏愛要度檢測，同時(shí)與普通PCR進區知(jìn)行敏感性比較；使用制備的探針，對(科玩duì)經(jīng)分離鑒定和制備保存的媽購其他7種(zhǒng)禽病毒核酸進(jìn)行特異性檢測；那低用該核酸探針，對(duì)疑似感染APV的鹦鹉病料進(jì理物n)行斑點雜交檢測，并對(duì)鑒定爲那算陽性的APV進(jìn)行全基因組擴增和序列謝朋分析。結果顯示：該探針可檢測到2 pg量的APV票年特異性核酸片段；僅APV-VP1陽性核酸顯色，呈現陽性反應，樂愛而陰性核酸和其他7種(zhǒng鐵輛)禽病毒核酸均不顯色，呈陰性反應。結果表明：建立的核酸斑點雜畫村交檢測方法具有較高的靈敏度和特異通南性，可用于臨床初步診斷。本方法的建立爲我國(guó問快)開(kāi)展APV分子流行病學(xué)調查及其感染的臨床診斷提供路會了技術支撐。Establishment and Applicatio樹著n of PCR Combined with Dot Blot 輛能Hybridization for 電會Detecting Avian PolyomavirusBudger國費igar fledgling disease（BFD）is an光白 acute viral infectious disea金資se caused by avian po笑文lyomavirus（APV），whic報歌h could lead to death of various parro林跳t nestlings and seriously endange哥我r the healthy development of par南舊rot industry. In order to improve劇友 the sensitivity and spec吃中ificity of molecular biologi大去cal methods to detect APV，the APV ge放唱ne fragment was c樹們loned and sequenced. After 讀來designing and synthesizing a 輛市pair of specific primers a林事s well as carrying ou喝做t PCR amplification森在，gene fragment with the nu在土cleotide length of 廠一731 bp was obtained by taking VP1 ge信化ne as a template，and then the fragment 和聽was marked with DIG to prepare a sp事煙ecific probe for detecting APV；the 動現sensitivity of the probe was detec舞場ted and compared with general P音船CR；other 7 kinds of i他站solated and prepared virus nuclei業笑c acids were us書明ed to evaluate the spec日喝ificity of the probe；the lesion吃少s of parrots suspected t舊件o be infected with APV were detected 了飛by dot blot hybr快小idization using the錢道 probe，and the whole genome sequen人煙ces of the positiv費廠e APV were amplified an刀金d analyzed. The 影中results showed that the prob不森e could detect 2 pg of APV specifi男唱c nucleic acid fragment；a雪村nd only the positive nucleic acids 商快of APV-VP1 showed posi海媽tive reaction，whi小黃le the negative one and other 7 kind輛化s of virus nucleic aci我吃ds all showed negative 現藍reaction. In conclusion，the establ數制ished method could be used for cl說們inical preliminary diagnosis d生開ue to its high sensitivit體影y and specificity. Th個微erefore，molecular epidemiological inves場子tigation and clinical diagno讀這sis of APV infection in C物會hina were provided with technical supp些亮orts by the established下化 method. 全文下載鏈接：https://k錢外ns.cnki.net/kcms/detail開對/detail.aspx?dbcode=CJFD&dbname=CJFDA又章UTO&filename=ZGDW姐少202103019&v=2vpJqQNi66ENu4TMbnp5wITes會大OgSWVYX3GNv8UqvwAK%25mmd2FHFW看購9r%25mmd2BJ4RMfJbfgtfmNo
2021-03-16
七類消毒劑對(duì)非洲豬瘟病毒熒光定量P錯高CR檢測結果的影響
爲評價戊二醛、酚、含碘類等常用消毒劑消毒後(hòu)對(什又duì)非洲豬瘟病毒熒光定量PCR檢測結果的影歌醫響，基于畜禽欄舍、運載工具、器具消毒及皮膚黏膜消毒目的，按消毒劑說(shuō算要)明書推薦選擇不同工作濃度，分别與不同滴度的非洲豬瘟病毒培養物于20 ℃條草店件下作用30 min後(hòu)，采用熒光定量PCR方法檢測作用後(hò但煙u)産物。結果顯示，與對(duì)應的陽性對(du遠動ì)照組相比，含氯類（二氯異氰尿酸鈉）、過(guò)硫酸氫鉀類、二氧化氯類消年些毒劑，消毒後(hòu)對(duì)熒光定量PCR檢測結果影響最拿慢顯著，檢測Ct值顯著上升或檢測不到；戊二醛類、含碘類（主要成(ché時理ng)分聚維酮碘）消毒劑，核酸降解能(néng)力相對(duì)較弱弟到，檢測Ct值稍有上升；酚類、季铵鹽類、含碘類（主要綠舊成(chéng)分碘、磷酸、硫酸）類消毒劑，檢測Ct值基本無變化。本研究評費問價了7類常用消毒劑消毒對(duì)非洲豬瘟病毒熒光定量PCR檢測結媽算果的影響，可爲防控實踐中科學(xué)、客觀評價分析消毒效果提供技術參考。T議書he Influence of Seve坐和n Categories of 和花Disinfectants on the Test Resu子務lts of ASFV by the Fluores兵數cent Quantitative P店的CRIn order to evaluate the in林請fluence of common disinfectan行頻ts including glutaral那人dehyde，phenol and io車員dine on the test results of African s會路wine fever virus（ASFV），the d自樹isinfectants with different concentr票那ations were used according to the 喝看corresponding instructions and based行懂 on the purpose of disinfecting筆冷 pens，vehicles，instrum化場ents and skin mucous mem學爸brane to react respectively with the 生草cultures of ASFV with different titer白民s at 20 ℃ for 30 min，th放服en the products were tested by畫說 fluorescent quantitative PCR. T大場he results showed that，compared to the 術去positive control group，t關費est results of using chlorine（sodiu化愛m dichloroisocyanurate），potas老門sium bisulfate and chlorine dioxide從西 compound disinfec慢妹tants were significantly affe姐靜cted，the Ct value signif車農icantly increased or failed to be detec你從ted；for glutaraldehyde and iodine c物年ompound disinfectants（mainly北照 contained povidone iodine），th她什e capability of nucleic acid吧如 degradation was re雜匠latively lower，and the Ct value slight說商ly increased；for phenol，quaternary ammo木到nium and iodine compound disi呢行nfectants（mainly contained iodine，音文phosphoric acid and su看山lfuric acid），the Ct value generally re如她mained the same. In conclusion，the in白腦fluence of seven categories 知科of disinfectants on the te小科st results of ASFV by fluorescent qu可路antitative PCR was少理 evaluated，which could provide some tec去開hnical reference暗兵s for scientifi文海c and objective evalua和動tion and analysis of disinfection e說計ffect in practice.全文下載鏈接：https://kns.cn理冷ki.net/kcms/detai跳又l/detail.aspx?dbcod區外e=CJFD&dbname=CJFDAUTO&filename=ZG妹討DW202103018&v=2vpJqQN化電i66F8aSTvSBXTPV1Y3MmmScXJxElc9間喝z63bfIKqOPCK4wm6NDvdOQjdWxT
2021-03-16
畜禽産品中四環素類藥物殘留色譜質譜檢測購得技術及前處理方法研究進(jìn)展
四環素類藥物是一類廣譜抗菌藥物，在畜牧業中得到東很廣泛應用。該藥物的不合理使用甚至濫用，導緻環境及動物源性食品中此類我他藥物殘留超标，從而威脅公共衛生安全。液相色譜法及液相色譜串聯質譜法是姐的檢測畜禽産品中四環素類藥物殘留的主要方法。這(zhè)類方法的難點及關鍵步驟弟志是樣(yàng)品的前處理，也是目前研究的熱點。本文就(jiù)目前動物源性食子員品中四環素類藥物殘留色譜及質譜分析法，特别是樣(yà風愛ng)品前處理方法的研究進(jìn)展進(jì討能n)行綜述。近年來新淨化材料、新提取劑及新提取方法不斷出現，提海媽升了提取及淨化效果，減少了試劑用量，提高了工作效率，并能(néng)夠同時見長(shí)實現多類藥物的提取和檢測銀暗，有力保障了食品和環境安全。Research Prog線技ress on the Detecti光錢on of Tetracycline Resid拍信ues in Livestock and Poultry Products b坐低y LC-MS and Study on Pre-書短treatment MethodsTetracyclines，a cl街金ass of broad-spectrum antibioti線門cs，has been widely a去樂pplied in animal husbandry. However，th見友eir residues in env讀離ironment and animal-d街美erived food would exceed the standard朋匠 in the event of any un哥文reasonable or abuse use，thus po雨事se a threat to public health safety上跳. Liquid chromatography（LC）an舞日d liquid chromatography-mass spe這自ctrometry（LC-MS）were the 農大main methods to detect tet聽哥racycline residue舞雨s in livestock and poul些快try products，and their difficu紅是lty and key step was pr媽刀e-treatment of samp你店les that was also a hots爸影pot under study. In少中 the paper，the LC and 術請LC-MS for detection of tetracycl腦個ine residues in animal-derive房小d food，especially the research人訊 progress of sample pre-treatment me玩林thods，were summariz術答ed. With the emerging new purific下件ation materials，extraction agents and 文黃methods，the safety of food 門老and environment wa鐘能s strongly safeguarded through imp他但roving the effectiveness of extrac高刀tion and purification，reducing 科話the dosage of reagents，increasing w制刀ork efficiency as 湖醫well as simultaneously extracting算校 and detecting multipl西金e drugs.全文下載鏈接：https熱訊://kns.cnki.net/kcms/det視月ail/detail.aspx?dbcode=CJFD&dbnam笑不e=CJFDAUTO&filename去高=ZGDW202103017&v=2vpJqQNi66HD98xa多又KRkUCe2%25mmd2FvY%25m友內md2BdoLqNpkBm9OA0N3AHjQbEicL7kr那紅HGw89vj4Hl
2021-03-16
非洲豬瘟自然弱毒株演變進(jìn)程
自2018年非洲豬瘟（African swine fever，ASF服慢）傳入中國(guó)以來疫情得到了有效控制。但2021年以購鄉後(hòu)，我國(guó)的ASF流行情況出現對服了新的變化，臨床中出現了“自然變異株”。這(zhè)些毒株可能(nén樂道g)導緻感染後(hòu)臨床症狀不典型，容易與其他疫病混淆等問題。山票這(zhè)與國(guó)外的流行演變規律一緻，即當ASF在一個國(g車區uó)家或地區流行較長(cháng)時(shí)間後(hòu)，其開看臨床表現將(jiāng)由急性發(熱道fā)病轉變爲緩慢發(fā)病內相或出現新的臨床表現。經(jīng)過(guò)多年的觀察積累，國(匠河guó)外已經(jīng)對(duì)該病毒的流行規律特别是自然弱毒株的演變愛腦進(jìn)程有了大量記載，但國(guó)内目前尚無這(zhè)方面(miàn來林)的描述。爲此，就(jiù)ASF自然弱毒株的演變進(j煙區ìn)程進(jìn)行綜述，從而提出加強血清學(xué)診斷産品研高冷發(fā)、加快産業化報批進(jìn)程、适時(shí)開(kāi)展血清學(答舞xué)監測追溯等建議，以期爲我國(gu們化ó)ASF科學(xué)防控提讀路供參考。An Overview of Evo你他lution Concerning Natural Att匠少enuated Strains of ASFVThe outb媽討reak of African間信 swine fever（ASF刀老）has been effectively con光哥trolled since African swine fever vir為歌us（ASFV）was int吧多roduced into Ch友明ina in 2018. But its prevalence飛草 has changed since 2021，tha火飛t is，“natural varia開美nts”are clinically 人車emerging. The strai大可ns may cause atypi好音cal clinical symptoms，and吃自 could be confused with other a風海nimal diseases. All of which is consis也了tent with the evolution rule in ot體呢her countries，that is，when美路 ASF is prevalent in a country or re暗紅gion for a long time，it秒報s clinical symptoms wil低外l transfer from 物事acute morbidity to slow occurrence o花視r new symptoms are 的舞emerging. The prevalence rule of劇信 the virus，especially the n做自atural attenuated st理路rains，has been large請水ly documented in黃地 other countries，except 體答China. Therefore，the evolution proces個物s of the natural a知這ttenuated strains was summarized 了理in the paper，and some su在錢ggestions were hereb了熱y put forward，such a光刀s further developing ser少暗ological diagnosis products，spee醫唱ding up the approval proc音拍ess for industrialization as well as c作美arrying out serological su拍去rveillance and tracing as鐵雨 appropriate，wi麗離th a view to prov海熱iding some references for 分那scientific preven愛短tion and control of ASF in China.全文下載費城鏈接：https://kns.cnki.net/kcms/detail/det資答ail.aspx?dbcode=CJFD&dbname=CJFDAUTO&信金filename=ZGDW202103015化從&v=2vpJqQNi66GkJtuoa4GJnJM%25m高離md2BMvT6D%25mmd船開2Fr5%25mmd2FMlD高討Syjfqrg20KKTyGmToW8Ifn物小3RGbxw
2021-03-16
動物新型冠狀病毒流行現狀
新型冠狀病毒（SARS-CoV-2）感染導緻的新冠肺炎（C微睡OVID-19）疫情自2019年12月以來，已在全球算議221個國(guó)家和地區傳播，導緻1億多人感染，230多鐘校萬人死亡。SARS-CoV-2除了對(duì)公共衛生産生腦請極大影響之外，全球已有24個國(guó)家和地區報告4門用70餘起(qǐ)動物感染疫情，涉及犬、貓、水貂、獅子湖動、老虎等多種(zhǒng)動物。因此，在同一健康框架下，應高度民廠關注動物感染SARS-CoV-2的早期預警。本文描述了動物感染SARS日的-CoV-2的國(guó)際流行特點，對(duì)其流行現狀和感染動物種友相(zhǒng)類進(jìn)行了綜合分析，評估了當前國(藍離guó)内防控動物SARS-CoV-2感染面(miàn)臨的形勢，件購并提出了針對(duì)性的防控建議，爲國(術答guó)内COVID-19聯防聯控和養殖業健康對如安全提供了借鑒。The Prevalence Status of S做關ARS-CoV-2 in AnimalsCoro票朋na virus disease 20木土19（COVID-19），caused弟秒 by severe acute respir件喝atory syndrome coronavirus 2（SA喝懂RS-CoV-2），has been spreadi了相ng in 221 countries and regions since 章器September 2019，leading to more東放 than 100 million persons infec物謝ted and 2.3 million 音地died. In addition to its c東民onsiderable effect 農兵on public health，more tha間快n 470 outbreaks in animals were also r視很eported in 24 co少做untries and reg來老ions，involving c的我anines，cats，minks，l弟船ions，tigers and other a光了nimals. Therefore，early warnin媽玩g of animal infec吧喝tion with SARS-CoV-2 should be但風 highly concerned un男鐵der the“one-health”framework. In the 自錯paper，the international preva愛花lence characteristics of SARS-Co林內V-2 in animals were described，the p開就revalence status and species 要我of infected animals were compr問爸ehensively analyzed，then the 懂會conditions for preventio分議n and control of SARS-C對師oV-2 in animals in China was evaluated，器腦and specific sugges高友tions for prevention a訊電nd control of SARS-CoV-藍低2 were put forward，h開鄉oping to provide 費錯some references for joint prev木短ention and control of the v暗姐irus as well as the health and safe街舊ty of livestock industry.全文下載民相鏈接：https://kns.cnki.net/kcms/detai為訊l/detail.aspx?dbcode=CJ很影FD&dbname=CJFDAUTO&filename=ZGDW2021030看多14&v=2vpJqQNi66HWgQ%25mmd2Bz聽吧2bqFmH%25mmd2BkG3SOxiQ%2討姐5mmd2B0l5c2HLERzR時西t0YyzkiRpu7LgOLgcxdxs
2021-03-16
RNA病毒重組及發(fā)生我國(guó)部分地區禽戊型肝炎病毒分子流行病學(線黃xué)分析
爲了解我國(guó)雞群中禽戊型肝炎病毒（avain h呢頻epatitis E virus，aHEV）的分子變異情況，對(duì少一)2018年從山東、河南、河北、遼甯、吉林、黑龍江、花金陝西、山西、江蘇等地疑似aHEV感染雞群中采集的679份肝髒、路樹脾髒病料樣(yàng)品進(jìn)行RT-PCR檢測，從中選取電刀PCR陽性産物，對(duì)其OR影些F1基因進(jìn)行序列測定與遺影下傳進(jìn)化分析。結果顯示：從病料中檢出陽性樣(yàng)品37份，音火陽性檢出率爲5.45%；選取的15個試驗毒株之間的ORF1基因同源性爲90.1黃體%~100%，與目前報道(dào)的4種(zhǒn有我g)基因型同源性均小于90%，其中與基因理亮 1 型的澳大利亞株和韓國(guó)株同源性爲85.5%~89.0%，房我與基因2型的美國(guó)原型株和美國(guó)無毒株同源性爲雪務85.5%~88.2%，與基因3型的歐洲株和中國(guó)株同源紙理性爲86.3%~89.7%，與基因 4 型的匈花海牙利株和中國(guó)台灣株同源性爲86.4森拿%~88.5%。ORF1基因進(jìn)化樹分山厭析發(fā)現，所分離的aHEV均屬于戊型肝炎B種(zhǒng)，不在議了目前所劃分的基因型分支上，形成(c間事héng)獨立的基因分支，不屬于已報道(dào)的4個基因型。結果多技表明，目前在我國(guó)流行的aHEV是一種(zhǒng)新基因型病毒長空，這(zhè)爲我國(guó)aHEV分子流行病學拍白(xué)分析提供了補充和依據。Molecular Epidemio得志logical Analysis on Avian Hepat人廠itis E Virusin Some Regions of ChinaIn商章 order to investigate the varia們日tion status of avian hepatitis E virus計討（aHEV）in chicken in China，679 lesi雜畫on samples of liv如頻ers and spleens collected from the chic化要ken suspected of being infected with 船數aHEV in Shandong，Henan，Hebei動白，Liaoning，Jilin，Heilongjiang，Shaanxi，S船微hanxi，and Jiangsu provinces in 201嗎秒8 were tested by RT-PCR，fr家吧om which，positive s懂和amples were selected，and th年煙eir ORF1 genes were sequenced and anal風要yzed for genetic evoluti紙吧on. The results showed that 37 positi司知ve samples were detected out服行 with the positive rate of 5.45%；the 國呢homology of ORF1 genes among 1森媽5 selected strain火樂s ranged from 90.1行也% to 100%，and the homology with th書日e four genotypes previously repor到路ted was lower than 90%，specifica農問lly，that with genotype 1（Australian and唱音 the Korean strains行風）was from 85.5% to 89.0%，that with the 照可genotype 2（American 西和prototype and avirulent輛村 strains）was from 85.5% to 裡女88.2%；that with the 錢關genotype 3（European and Chine風就se strains）was 老黃from 86.3% to 89.7%；and that 行區with the genotype 4（Hungari讀書an and China Taiwan strains）was from 8但費6.4% to 88.5%. It was f頻習ound that，by phylogenetic tree an員就alysis of ORF1 genes，all the isolated a窗車HEV strains belonged to g拍個enotype B of hepatitis E，wh下說ich was not locat拿鐘ed in current genotype bran區業ch，but a new independent br新離anch outside of t熱得he four reported ge花上notypes. In conclusion，t去街he aHEV strain pr厭一evalent in China was你術 a new genotype vi樂的rus，which provided a supplement 藍也and basis for molecular epidemiologic員廠al analysis on aH日書EV.全文下載鏈接：https://kns.頻呢cnki.net/kcms/detail/detail.asp東文x?dbcode=CJFD&dbname=CJFDAUTO&f就家ilename=ZGDW202103001&v=2vpJqQNi6歌鄉6EYoeXG8NysKNTLG%25mmd2BSLPA作中V%25mmd2F9kJo6Jm0o少長xCBbOGcNz8zXyVXnrg3店中VoTk
2021-03-16
規模豬場密閉連廊及相關設施的設計和應用
自2018年非洲豬瘟傳入我國(guó)以來服了，生豬養殖業遭受了沉重打擊。建設規模豬場密閉連廊，些睡可有效切斷非洲豬瘟等動物疫病的傳播途徑。廣西橫縣通過(guò)設計和應用密閉雜算連廊連接生活區、豬舍、物資熏蒸房雜報、出（進(jìn)）豬台及死豬出口等端器跳口，使豬舍防疫關口前移，增加了防疫縱深，實現了多層阻斷抵禦病原傳播。本文介了多紹了密閉連廊及相關設施的設計要點和使用流程，以期爲規模豬場落實上風各項生物安全措施，防控非洲豬瘟等動物疫病提供參考。Design業樂 and Application 歌畫of Enclosed Corridors and Related Fac從媽ilities in Scale Swine開遠 FarmsPig production industry h日照as been seriously damaged by Af線票rican swine fever（ASF）i費畫n China since 2018 when it was introd明窗uced into China.場場 The spreading routes of animal di討通seases including ASF資不 could be effec老匠tively blocked through the construction說森 of enclosed corridors in scal紙煙e swine farms. In Heng County of G河農uangxi，the living area，pens，material 慢資fumigation room，load線什ing/unloading platform，d算裡ischarge outlet f購就or dead pigs and other ports we照短re connected by the enclosed corr用開idors，by which the ga友藍teway to pens for 可飛disease prevention and c間道ontrol was moved河關 forward to go deep into the議購 prevention and control me訊熱asures，so as to block the spread of pa媽月thogens through such multi-layer bloc費花king. In the paper，the d城放esign considera樹為tions and application process懂船 of enclosed corridors and r明開elated facilities were introduced fo關從r the purpose of pro多嗎viding some references for 是離implementation of various bio-secur綠近ity measures，prevention and control 笑但of ASF and other animal d對裡iseases in scale farms. 全文下事討載鏈接：https://kns.cnki.net/kcms/detail/和家detail.aspx?dbcode=CJFD&dbname=CJFDAUTO生術&filename=ZGDW202102014&v=2vpJ腦的qQNi66EkqWZmAe7NlkUZ08tJo2iW7N0Pl6G自看HCHEh%25mmd2FGT1fy快山Ltm0bdV5pRcOW%25mmd2F
2021-02-26
超高效液相色譜-質譜法測定豬尿中15種(zhǒng)β-受體阻斷路區劑殘留
爲建立一種(zhǒng)快速測定豬尿冷商中15種(zhǒng)β-受體阻斷劑殘學也留的超高效液相色譜-質譜（UPLC-MS/MS業子）确證方法，本試驗優化了分子印迹固相萃取（MI暗要SPE）淨化條件及色譜、質譜條件。豬尿樣(yà白資ng)品經(jīng)離心後(hòu)加入線年MISPE柱上樣(yàng)，依次經(動店jīng)水、乙腈淋洗，10%乙酸甲醇洗脫，N2吹幹，0.1%甲兵美酸水-乙腈（v:v=9：1）複化了溶；采用ESI正離子模式在多反應監測模式下進(jìn)船銀行離子掃描，外标法定量。結果顯示，豬尿中15種(zhǒng)藥物在0.1爸吃~10 μg/L範圍内具有良好(hǎo)的線性關系（R2≥0.996）喝訊，檢出限和定量限分别爲0.03和0.10 μ房小g/L；各藥物回收率在76.7%~10土書9.2%之間，批内、批間變異系數分别在2.2%~18.7%、2.1%~1南子9.7%之間。結果表明，本方法操作簡單快速、靈敏度高放開、特異性好(hǎo)。Determination問木 of 15 β-blocker Resi視裡dues in Pig Urine Using Ultra-頻可high Performance Liquid Chromatograp要女hy-Mass SpectrometryIn order 都線to establish an ultra-high performanc男坐e liquid chromatogra那務phy-mass spectrometry（UPLC-MS/MS）to 草商rapidly determine 15 坐離kinds of β-blocker residues 短腦in pig urine，the conditions林到 for purification of molecular imprinte業家d solid phase extraction（MISPE看舊）as well as chromatograp物他hic and mass spectrometry we錢間re optimized in the study. Pig ur就請ine samples were c不暗entrifuged and 問睡added in the MISPE colum木跳n，then rinsed wit白書h water and acetonitrile successively，e在藍luted with 10% ace是老tic acid methanol，dried with 來畫N2，and redissolved with 0.1%土弟 formic acid water aceto懂為nitrile（V:V=9:1）；ion scanning was carr司美ied out by ESI positive ion mode 們什under multi-reacti商的on monitoring mode（MRM），whi那她ch was quantified by external裡舊 standard method. The re民舊sults showed that the 時煙15 drugs were with a good linear去雜 relationship wi謝鐘thin 0.1~10 μg/L（R2≥0.996），and th來計e limits of detect金山ion（LOD）and quan女些tification（LOQ）were 0.03 and 0.10 商你μg/L，respectively；the recovery rate of 都計all the drugs ranged from 76.7% 近動to 109.2%，and the coef吧資ficients of vari也關ation ranged 2.2%~18站那.7% and 2.1%~19.7習刀%，respectively. It was con近數cluded that the method 錢哥was simple，rapid，se時都nsitive and specific.全文下載鏈接：https://k民影ns.cnki.net/kcms/detail/de看銀tail.aspx?dbcode=CJ睡們FD&dbname=CJFDAUTO&filenam爸公e=ZGDW202102025&v=2v個校pJqQNi66GojmWBc姐女c%25mmd2B6miRpG%25mmd2Bo8GsLrhOfHgYqN到離YW69EExMBPYDl2RICFk9Kmq4
2021-02-26
三株牛支原體的緻病性比較
爲檢測從内蒙古發(fā)病牛場分離到的3株牛說好支原體（HS2019、HSZ2019開去、HSS2019）的緻病性，對(duì)其進(jìn)行本動物回歸試驗，通妹通過(guò)觀察攻毒後(hòu少呢)的臨床症狀、病理變化，以及應用實時(shí)熒光定量PCR确定組織器官中科妹支原體載量，分析3株支原體的毒力。結果顯示，3株牛支原體回歸牛長好體後(hòu)均使試驗牛出現體溫升高、咳嗽、呼吸困難等臨床症狀，解剖可觀察到試就務驗牛肺部損傷以及肺髒與胸腔粘連的病理現東變化，其中HS2019株較其他兩(l術工iǎng)株引起(qǐ)的症狀與病變更爲明顯，通能組織髒器中的載菌量最高。結果表明，這(zh坐林è)3株支原體均有緻病性，其中HS2019株緻病性最強，可作爲今後(hòu訊討)疫苗研制的預備菌株。本研究既爲國(guó)内疫鐘黃苗的研制提供了菌株資源，也爲今後(冷你hòu)牛支原體的免疫攻毒試驗提供了評價标準。Comp習森arison of the Virulence of Three雨中 Strains of Mycoplasma bovis In order 玩兒to test the virulence of three stra窗睡ins of Mycoplasma bovis（HS2019，HSZ2019 雪又and HSS2019）isolated from an infected做鐵 farm in Inner Mongolia，an ani林我mal regression test was carried out河身，the number of Mycoplasma in tissue an子農d organs was determi外中ned by observing低著 clinical symptoms and pathologica就暗l changes after virus chal熱我lenge as well as by real-time flu熱作orescent quantitative PCR，then 頻在the virulence of the得愛 three strains was 南鐘analyzed. The results sho年資wed that such clinic技南al symptoms as 高站increasing body temperature，c黃暗ough and dyspnea appeared in exper工著imental cattle after 西慢the regression te錢樂st of the three 到一strains. The lung le行體sions as well as adhes火一ion between lung and thora鐘歌cic cavity and 就熱other pathologi路場cal changes could be observ頻看ed after anatomy，es行花pecially HS2019讀匠 strain，by whic不匠h more obvious symptoms an員這d lesions were caused，and生理 the volume of Mycoplasma in tissues 吧明was maximum. In con湖又clusion，the three strains of Mycoplasm用間a were all pathogeni哥要c，especially HS2019 strain who我技se virulence was the stronges藍爸t，which could be used as the preparatio弟湖n strain for vaccine deve下水lopment in the future. By the 土黑study，strain resources wer愛吃e provided for the development of麗微 vaccines，and evaluation criterion f一亮or immune challenge test o睡花f Mycoplasma bovis in the f務這uture was also p分著rovided.全文下載鏈接：https://kns.上花cnki.net/kcms/detail/detail.aspx?dbcode地那=CJFD&dbname=CJF能木DAUTO&filename=ZGDW202102023&v=2vpJq你關QNi66FfOvHvmWPuBUnH路票9uxLqtYVTp8wOiuUbXQ9dmsl年用aZtj8%25mmd2BtumdMbS7yB
2021-02-26
裂谷熱病毒Gn蛋白主要抗原區的串聯表達和地高多克隆抗體制備
裂谷熱（Rift Valley f一土ever，RVF）是由裂谷熱病毒（Rift拍舊 Valley fever virus，RVFV）引起(qǐ)身人的一種(zhǒng)烈性人獸共患傳染病。R音道VFV囊膜蛋白Gn可誘導産生中那影和抗體，是RVFV檢測方法和疫苗研究的重要抗原靶标。件員本研究通過(guò)分析蛋白抗原位點信息，構建包含Gn蛋白兩(liǎ愛見ng)個主要抗原區域的重組表達載體，随後(hòu)將(jiāng)質微購粒轉化至BL21感受态細胞，以IPTG誘導要購重組蛋白表達并優化蛋白表達條件，通過(guò)Western Blot鑒定重他水組蛋白；將(jiāng)重組蛋白免疫BALB/c小鼠，制備多克隆抗體，開老并以ELISA、Western Blot、IFA檢測多克隆抗店好體的反應性。結果顯示：誘導表達的Gn重組哥子蛋白分子質量約爲45 kDa；蛋白表達條件優化爲IPTG終濃度0.25 謝遠mmol/L，誘導時(shí)間5 h；Wes大區tern Blot鑒定發(fā)現蛋白成(chéng)功視兵表達。通過(guò)ELISA測定小鼠三免後(hòu)血清抗體，結果發(fā)體飛現抗體效價大于1:51 200；We音高stern Blot檢測顯示，制備的多抗血清能(n睡弟éng)與重組蛋白發(fā)生反應；進(jìn)一步的做她IFA檢測結果表明，制備的多克隆抗體可與真核質粒轉染細胞中表達的Gn蛋白花章反應。本研究獲得的Gn重組蛋白及制朋拿備的多克隆抗體爲後(hòu)續RVFV檢測方日劇法的建立奠定了基礎。Tandem Express下弟ion of the Major到飛 Antigenic Areas of RVFV Gn P頻事rotein and the Preparat湖高ion of Polyclonal Antibodies Rift V唱自alley fever（RVF）is a virule會木nt zoonotic disease caused by Rift行媽 Valley fever virus（RVFV）. The envel湖哥ope protein Gn of RVFV is an imp做你ortant antigen 外年target for the detection method develo問唱pment and vaccine resea劇站rch，as it could induce the production 著西of neutralizing antibodies. In the stud東得y，the recombinant expressio厭通n vector containing two maj這關or antigen areas裡好 of Gn protein was constructed through 木務analyzing the inf那木ormation of prot市到ein antigen sites，then 輛雪the plasmid was transformed into BL21請你 cells，the recombinant protein expre能說ssion was induced by IPTG，followed by 麗靜optimization of relev飛算ant conditions，the recombinant 市麗protein was identified by W線車estern Blot and then vaccinated 兵會into BALB/c mice to prepare the 窗好polyclonal antibodies that were 問一tested by ELISA，Western Blot 電會and IFA. The results sh就她owed that the molecular weight o看紅f the induced Gn recombinant protei街樂n was about 45 kDa；the exp用作ression condition湖機s were optimized as follows：the final 錢醫concentration of IPTG was 0.25 mmol/L 麗要with the induction time of 5 h；and the金老 protein was successfully 新木expressed as identified by Weste一下rn Blot. The antibo生會dy titer in vaccinated mice was higher唱新 than 1:51 200 城河as tested by ELISA；it was shown by W會輛estern Blot that，the pr我懂epared polyclonal antiserum coul快相d react with the歌放 recombinant pr舊討otein；and the polyclonal ant身吧ibodies could react with the Gn 問票protein expressed in雨們 eukaryotic transfection外和 cells as detected by 訊公IFA. In conclusion，future d錯那evelopment of RVF detect遠弟ion methods was provided with藍森 a basis by the expression of Gn 工裡protein and preparatio街視n of polyclonal antibodi信河es. 全文下載鏈接：https://kns.c山麗nki.net/kcms/detail討計/detail.aspx?dbcode=CJ看來FD&dbname=CJFDA吧報UTO&filename=ZGDW什票202102022&v=2vpJqQNi66GGZ林弟3UELIBPZGPh607vmumyimku快綠iumm5ssFBue57bzwY4ZSGwS2F草個GnC
2021-02-26
1型、2型牛病毒性腹瀉病毒通用RT-PCR檢測方法的電窗建立與應用
爲建立一種(zhǒng)快速檢測1型、筆員2型牛病毒性腹瀉病毒（BVDV錯關）的通用RT-PCR方法，根據GenBa算科nk上收錄的64株1型、2型BVDV以及1株豬瘟病毒（CSFV）的全基又能因組序列，應用Primer 6.0軟件設計林可針對(duì)5'-UTR區域的1型、2型看如BVDV特異性通用引物對(duì)，店又擴增目的片段，并對(duì)該方法進(jìn)行特異站裡性、敏感性、重複性試驗及利用該方法開(兵頻kāi)展臨床樣(yàng)品檢測。結果顯示：擴增的目的片段長(ch空睡áng)度約爲302 bp；該方法的靈敏錢會度爲2.09×102 copies/μ弟自L，無非特異性擴增，且重複性良好(hǎo)。對(duì)采自山東省發(fā友農)病牛場的13份鼻腔棉拭子和9份牛血清臨床樣(yàng)品進(習我jìn)行檢測，發(fā)現有8份鼻腔棉拭門明子和8份血清爲BVDV陽性，随機訊日抽取4份陽性樣(yàng)品送測序，發訊視(fā)現3份樣(yàng)品毒河照株爲1型BVDV，1份樣(yàng)品還(há機刀i)需進(jìn)一步鑒定。本研究建立的RT-PCR方飛場法可實現對(duì)1型、2型BVDV核酸的特錢黃異性檢測，也可用于該病的流行病學(xué)調老見查研究。Establishment and Application 友計of a General RT-PCR Assay for嗎民 Detection of BVDV Type 1 and 2In orde女靜r to establish a general RT-PCR assa習路y to rapidly det影風ect bovine vira空拍l virus（BVDV）type 1 and 2，the general p也會rimer was designed based on 5&房新#39;UTR sequences師體 of BVDV type 1 and 2 by Pr是哥imer 6.0 software according 不窗to the analysis of complete gene se她著quence of 64 strains of BVDV type裡開 1and 2 and one strain of classical銀通 swine fever virus（CSFV）r呢你egistered in GenBank，an女黃d the target fragm體房ent was amplified，the說朋n the specificity，sensitivity and repro業聽ducibility of the RT-PCR me南行thod were evaluated，頻有and the clinical samples were tested說為. The results showed that the 西飛amplified target fragment was abo算中ut 302 bp；the detection limt of the醫資 method was 2.09×102 copi線書es/μL，with no nonspecific amplif光答ication but good reproducibil市知ity. 13 nasal swabs and 9 bovine serum 鐵近samples collected from an 店訊infected farm in Shandong Province下遠 were detected by the esta開計blished method，it w裡訊as found that 8 nasa紅習l swabs and 8 sera were positive agai森吧nst BVDV，then 4 of the positive sa請謝mples were randomly selec輛窗ted to carry out virus gene sequencing哥紅 and comparison，it wa科在s found that vir低鐘us of 3 samples could be ident也時ified as BVDV type 1，and the ot報員her 1 sample still needed further anal什民ysis. In conclusion，spe舞靜cific detection of 明制nucleic acids of BVDV 1 and 2 co化不uld be realized 新店by the RT-PCR assay establ這他ished in the study，w兒請hich could also be used 笑坐in epidemiological investigati銀會on of BVDV.全文下載鏈接：老離https://kns.cnki.net/kcms/detail/det物區ail.aspx?dbcode=CJFD&dbname=CJFDAUTO&讀著filename=ZGDW202102020&v=2vpJqQ雜說Ni66ECPxin0tV%25mmd2Bw光鐵XuEDdY2ncWQTI3QnKz94n6p6pDeO9uI1裡大raFF86me8bE
2021-02-26
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