豬急性腹瀉綜合征病毒RT-PCR檢測方法的建立裡來及應用
爲建立一種(zhǒng)鑒定豬急性腹瀉綜合征病毒（SADS-時物CoV）的RT-PCR方法，根據NCBI登錄的S風東ADS-CoV N基因保守區域序列，設計了3對(duì)引物（美生P1、P2和P3），通過(guò)對(duì北坐)SADS-CoV及其他6種(zhǒng)病毒進(jìn)行檢測，篩選出最畫雨合适的引物，然後(hòu)對(du土河ì)退火溫度、引物濃度、dNTPs濃度、rTaq DNA聚合酶濃度請微和循環次數等反應條件進(jìn)行了優化。結果顯示，P3引物厭高對(duì)SADS-CoV特異性最好(hǎo)，與些玩其他病毒無交叉反應。敏感性試驗顯示，該方法最低檢測限可達呢我9.55×102 copies/μL；重複影件性試驗顯示該方法重複性良好(hǎo)。運用建立的RT-PCR嗎謝方法，對(duì)廣東省的21份臨床樣(yàng)品進(jìn)行檢測，舞討結果檢出SADS-CoV陽性樣(yàng)章謝品4份，陽性樣(yàng)品的測序結果與其RT-PCR完全一緻。結果表木愛明，本研究成(chéng)功建立了一種(zhǒng)鑒定SADS-CoV的很愛RT-PCR方法，且該方法具有檢測快速、成(chéng)飛飛本廉價、特異性強、敏感性高和重複性好(hǎo)的優點，可以用多行于SADS-CoV的臨床診斷。Establish鐘線ment and Application of a錯可 RT-PCR Assay for Detection o又著f Porcine Acute Di章東arrhea Syndrome Coronav跳行irusIn order to establish a RT-PCR as問制say for detection of porc拿事ine acute diarrhe服金a syndrome virus（SADS-CoV），three p路農airs of primers（P1，P2鐘匠 and P3）were designed bas看鐵ed on N gene sequence of SA我明DS-CoV registered in NCBI，and the most 請水appropriate primer was selected 姐兵through the detect家器ion of SADS-CoV an離頻d other six kinds of viruses，follow科麗ed by the optimization of reaction些數 conditions including annealing tem聽物perature，primer concentration舊時，dNTPs concentrati水站on，rTaq DNA polymerase c黃朋oncentration，cycle index，etc.動志 The results showed that P3 pri城房mer was with the best specificit海都y against SADS-CoV，and f場銀ailed to react with 匠北other viruses. 頻熱It was shown that金錢，by sensitivity test，the lowest detect間兵ion limit was 9.55×10熱歌2 copies/μL；the established assay was w美睡ith good repeatability as tested. 21 和報clinical samples from Guangdong were de麗紅tected by the RT-PCR ass訊家ay，4 positive samples were found，長頻which was consistent with t請數he sequencing res書報ults. Therefore，a RT-PCR一得 assay was successfully establi體知shed for detect科新ion of SADS-CoV，商妹which was chara好下cterized by rapid detection，拿線low cost，strong sp西員ecificity，high sensit開通ivity and good repeatability，and 事了could be used for identification of SAD能我S-CoV in practice.全文下載鏈接：https雪章://kns.cnki.net/kcms/detail/detail.理山aspx?dbcode=CJFD&dbname=C物票JFDAUTO&filename=ZGDW202011015&v=Bh8a8筆場ZT44%25mmd2BV3EfgMfB9vvEyWkN說關TMtTs%25mmd2BzGvsR畫說eoG5nuBXoFx9PNzPdOLT不船K1PFyhD
2020-11-06
環境DNA應用研究進(jìn)展
近年來，環境DNA（eDNA）應用發(fā)展迅速，已涉及食品微生物、生物也民監測、群落生态學(xué)、古環境研究、保護生物學(xué)從城和入侵生态學(xué)等多個領域。基于eDNA的研究子也手段具有非侵入性、易于取樣(yàng)和能(néng)夠探測隐秘物種(zhǒ頻西ng)等優點，因此eDNA是生态系統廣泛直接取樣(y下信àng)的一種(zhǒng)有吸引力的替代品。本文綜述了eDNA的定義月玩和發(fā)展曆史，介紹了DNA條形碼選擇和設計、樣(yàng)品收線文集、DNA抽提、DNA擴增測序、DN黃他A條形碼數據分析等eDNA研究步驟，描述了eDNA在外樂金來物種(zhǒng)入侵、物種(zhǒng工民)檢測、生物量估算、水中生态系統監測等領域的們笑應用，總結出eDNA研究中存在的問題，并展望該方法的應用前景。本文爲全面(m船請iàn)了解eDNA的應用提供了參考。 Research Progress多他 on the Application of Envi你場ronmental DNAIn recent years，enviro可地nmental DNA（eDNA）has been wi火南dely used in vario上的us fields，including food microb視還iology，biological monitoring，communi河票ty ecology，study on 雨樹palaeo-environment，con小我servation biology and invasion科從 ecology，etc. A木跳s the study methods of eDNA were 車問advantaged by non-invasiveness，convenie微好nce for sampling and d樹月etection of cryptic speci北到es，eDNA could be co銀商nsidered as an attractive alte街雪rnative to extensive dire雨笑ct sampling in ecological system. In th技河is paper，its definition and development線微 history were summarized；the stu行暗dy steps were introduced from the aspec木些ts of DNA barcode s山票election and design，sample col黃遠lection，DNA ext路如raction，DNA amplificati工銀on and sequencing，and anal議在ysis on DNA barcode data；then its ap做不plication in such fields as alien來日 species invasion光為，species detection，biomass 爸知estimation and aquatic ecos來訊ystem monitoring，年能was described；all problems encou筆很ntered during the above steps were sum做廠marized，and its 區路application prospect was prospecte劇但d. The review provided some re美月ferences to fully l懂學earn the application但個 of eDNA. 全文下載鏈接：劇新https://kns.cnki.畫制net/kcms/detail/detail.aspx?場舞dbcode=CJFD&dbname=CJ司姐FDAUTO&filename=ZGDW202011014&v=B煙視h8a8ZT44%25mmd2BXhkriUVQ308eGbsO7mbLu做坐gIY40POMzvPIr%25mmd2F7pVBhEQB7y信東CBgcfpV7P
2020-11-06
鹵蟲作爲生物餌料傳播蝦類病原風險的研究進(jì但年n)展
鹵蟲是蝦類育苗的關鍵性生物餌料，其是否具有傳喝爸播蝦類病原的風險已引起(qǐ)多文麗方關注。本文梳理了鹵蟲作爲生物餌料傳播蝦類病原的研究進(jìn)展，并提出了弟又防控建議。鹵蟲是白斑綜合征病毒（white spot syndrom道我e virus，WSSV）的機到來械攜帶者，也可能(néng)是對(duì)蝦傳染性肌壞死時了病毒（infectious myonecrosis virus，IMNV）、高窗肝胰腺細小病毒（hepatopancreatic parvovirus廠學，HPV）、羅氏沼蝦野田村病毒妹紅（Macrobrachium rosenbergii nodavirus，M哥那rNV）和極小病毒（extra small virus，XSV）的傳播鄉我載體，但是目前的研究由于缺少原位物相雜交或組織細胞水平的病理學(xué)證據，相照均尚未證實鹵蟲可被(bèi)這(z分鐵hè)些病毒直接感染；研究已證明鹵蟲可被(醫友bèi)副溶血弧菌（Vibrio parahaemolyticus）靜聽、坎貝氏弧菌（V. campbellii）還購和哈維氏弧菌（V. harveyi）等水産養殖緻病菌感染，存在傳播急車見性肝胰腺壞死病（acute h年哥epatopancreatic necrosis disease，AH中很PND）的風險。此外鹵蟲可能(néng)被(bèi)蝦肝腸胞蟲（Ente做雜rocytozoon hepatopenaei，EHP）污染唱技而存在傳播風險。因此，建議系統兵匠開(kāi)展鹵蟲的病原學(xué)和流行病學(討訊xué)研究，通過(guò)規範鹵蟲卵生産管理和孵化操作低吧、探索鹵蟲生物安保産業發(fā)展途徑和替代産品等措施，降低鹵冷放蟲傳播相關病原的風險。本文可爲正微得确使用鹵蟲等生物餌料提供技術支持，爲水産養殖綠色發(件中fā)展和生物安保體系建設提供參考。 Research Progr知玩ess on the Risk of Artemia Ac制購ting as a Kind of Live Fe請學ed to Spread Shrimp Pat我分hogensArtemia is a k近些ind of critical live feed國間 for shrimp breeding，whether i店女t could bring the risk of spre我章ading pathogens has been concern山的ed by the society. In this paper，the re國購search progress on Artemia acting as森樹 a kind of live 地到feed to spread pathogens 區窗was summarized，and relevant suggestio白低ns were put forward. Artemia served as 校劇the physical carrie頻城r for white spot syndrome viru舊木s（WSSV），as the vecto房風r carrying infectious myonecr友分osis virus（IMNV），he刀近patopancreatic parvovirus（HPV），Macrob票見rachium rosenbergii noda男動virus（MrNV），and extra smal訊國l virus（XSV）. However，作術current studies have not proved t黑的hat Artemia can be infected w開紙ith the above-mentioned vi木草ruses，due to luck of evidences of in s術音itu hybridization or 開裡other cyto-or histopathology你公. It was demonstr現門ated that Artemia can 愛嗎be infected with aqu樂畫aculture pathogenic bacteria，such 畫土as vibrio parahaemo樹開lyticus，V. campbell能員ii and V. harveyi，resulting in a risk 做南of spreading acute hepatopancreati還森c necrosis disease（AHPND）. Addit場又ionally，enterocytozoon hepat醫店openaei（EHP）may be tr車會ansimitted through Artemia by contamina鄉哥tion. Therefore，it was suggested to年黑 systematically carry o什中ut the study on the eti件高ology and epidemiology of artem關坐ia，and reduce the risk of s又煙preading pathogens through 快年standardizing producti水年on management or h藍知atching control of artemi匠農a cytes，identifying the biosecurity 木光pathways of artemia and devel笑議oping alternative products. Technic視坐al support could be prov冷劇ided to correctly use arte區員mia and other live feed and so坐謝me references wer紙鐘e provided for construction of a哥樹 biosecurity system 火的and for healthy develop機店ment of aquaculture.全文下載鏈接：https但化://kns.cnki.net/kcms/detail/d男訊etail.aspx?dbco雨裡de=CJFD&dbname=CJFDAUTO&filename=ZGD件海W202011013&v=Bh8a8銀子ZT44%25mmd2BXAoeiGBd4M8wAjjf78KMC0I讀但GJlzeY1p3prVG5rt1B7dRNsg妹老%25mmd2BjdZ%25mmd2FLC
2020-11-06
2016—2019年我國(guó)I畫視類新城疫病原學(xué)監測與遺傳進(jìn)化分析
爲掌握我國(guó)I類新城疫病毒（Newcastle件業 disease virus，NDV）的分布及分子演化特征，20技錯16—2019年，随機從國(guó)内23個省（自治區、直轄市）998個船船采樣(yàng)點，采集家禽口咽/洩殖腔雙拭子樣(yàng答空)本、野鳥糞便樣(yàng)本和環境樣(通林yàng)本共45 458份，通過(guò)病毒分離和RT-P山動CR鑒定，共分離到I類NDV 558株。從時(shí)間分布看，2016—20雪外19年I類NDV分離率分别爲0.95%、0.89%、1.88%和1.3亮雨7%，有增高趨勢；從地理分布看，我國是拍(guó)16個省（自治區、直轄市）分離到I類NDV，主要好在集中于華東、西南和西北地區；從場點分布看，活禽批發(fā)市雪月場和農貿市場的I類NDV陽性率票書明顯高于養殖場和屠宰場；從宿主分布看，I類NDV主要來源于雞和鴨，少量來自兵北鵝、鴿和環境樣(yàng)本；從基因型分布看，55場技8株I類NDV屬于2個基因亞型，其中1.1音物.2亞型爲近年來我國(guó)流行的樂制主要基因亞型。研究表明，我國(gu廠近ó)I類NDV污染面(miàn)廣，宿主範嗎木圍廣泛，且存在2種(zhǒng)基因亞型。研究提示，應加強I類NDV的免電影疫、監測，同時(shí)加強飼養管理，提高家禽抵抗力，降低病毒基因變異很冷緻毒力增強的風險。本研究爲我國(guó)科學(xué北訊)防控新城疫提供了參考數據。Pathogenic Sur了現veillance and Phylo媽物genetic Analysis on Class I Newcastle D呢得isease Virus du妹線ring 2016 to 2019 in ChinaIn 林頻order to study the distribution and 暗還molecular evolution chara腦哥cteristics of class I New時劇castle disease virus（NDV）in C業裡hina，a total of 45 4又務58 samples including avi不報an oropharynx/cloacal swabs很嗎，wild bird fece草知s and environmental samples were ra厭呢ndomly collected from 998 samplin玩作g sites in 23 province通司s（autonomous regions and munici廠河palities）from 2016 to 2019，and 558 長男strains of class I NDV w雨習ere isolated through virus is快如olation and RT-PCR. For time di河什stribution，the isolation rates 可麗from 2016 to 2019 we章房re 0.95%，0.89%，1.88% and 1.37%，respec都中tively，and tended to increase；fo快數r geographical distrib間員ution，such NDV strains were is討店olated from 16 provin林也ces（autonomous regions and 麗到municipalities），particularl要就y in East，Southwest an請事d Northwest China；for farms/pre要熱mises，the positiv友街e rate of NDV in live poul著鄉try markets and agricultura公有l markets was obviously higher t影農han that in farms 這請and slaughterhouse筆他s；for host distribution，the 如謝NDV strains were mai要知nly derived from chickens and d好從ucks，and fewer from冷區 geese，pigeons and environment；a照農nd for genotype distribution，the isol鄉亮ated strains fell into tw行我o genotypes，in which，1.1.2 subt光公ype was predomi服朋nant in China in recent y水遠ears. It was conclud些農ed that the NDV 有喝was widely spread wi弟草th a large range of hosts and 工他two genotypes. Therefo她校re，the vaccination and surveilla子著nce against NDV should be strengthened，我都meanwhile，feeding management should 機高be enhanced so a得靜s to improve relevant res笑視istance of poultry and reduce 物去the risk of increased virulence pro吧山duced by virus genetic variation. In vi劇草ew of the above，some reference d機術ata were provided for scientif多山ic prevention and control of NDV in Ch師農ina.全文下載鏈接：https://kns.cnki.n視工et/kcms/detail/detail.as頻月px?dbcode=CJFD&dbname術木=CJFDAUTO&filename=ZGDW20201100爸現1&v=Bh8a8ZT44%25m放家md2BUut1OAVi1CyFvtFbyONbzZR0Jl坐綠I3CIO8pIj5EAp%25mmd2B61e0YSoH%25mmd2F女說A7kcV
2020-11-06
豬繁殖與呼吸綜合征病毒GP5蛋白的原核表達及多克雨鐵隆抗體制備
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2020-10-14
豬瘟病毒C株檢測用國(guó)家核酸标準物質的研制
爲研制均勻、穩定的豬瘟病毒（CSFV）核酸标準物質銀門，選擇CSFV C株接種(zhǒng)多讀于PK-15細胞并收獲病毒液，然後(hòu)經(jīng)鄉哥過(guò)病毒滅活、分裝、凍幹，最終形成(c老近héng)核酸标準物質。用微滴式數字PCR對(duì)制備的CSFV标準物討靜質進(jìn)行均勻性和穩定性評估、标準物質合作定值，在評定不确定快弟度後(hòu)計算得出最終量值結果，并開(kāi)展臨床試用。暗房結果顯示，該标準物質均勻，4 ℃條件下可穩定存放4周，25 ℃可穩定存放著錯1周，-20 ℃可穩定存放11個月，公劇量值結果爲（5.1±0.7）×105 copie體厭s/mg；臨床試用顯示，所制備的标準物質與臨床樣(yàn黑雜g)本的互通性良好(hǎo)。本研究制備的CSFV C株國(g為電uó)家核酸标準物質均一性良好(hǎo)、穩定、量值準确度高，并從吧通過(guò)了全國(guó)标準物質管理委員會(huì)評定，可用于制服動物及動物産品質量安全檢測，從而有效保障豬瘟防治工作的開(kāi)路東展。Preparation of National Nucle答區ic Acid Reference Materials of CS事技FV C Strain for Detection In ord讀微er to prepare a輛章 kind of uniform and stable nu志著cleic acid reference新錯 materials of classical swine 風東fever virus（CSFV），CSFV C stra我制in was selected and inoculated int紙技o PK-15 cells to obtain viral fluid，f多草ollowed by the 遠木process of inact你電ivation，subpackage a靜城nd freeze-drying，then the 都道nucleic acid reference materials we科話re produced. The homogeneity and s遠地tability of prepared materials were ev看懂aluated by droplet digital PCR，and th影喝e value of materials was determined by 友件8 collaborative laborato筆算ries，the quantity value was ultimately 服作calculated after evalu做你ation of uncertainty，then a c林裡linical trial was c坐嗎arried out. The results showed that the學草 material was homogeneous and could be 好劇stored stably for four 報廠weeks at 4 ℃，for one week at 25 ℃，唱線and for 11 months業北 at -20 ℃，the quantity value was（著市5.1±0.7）×105 copies/放間mg；it was found that the mater內紙ials showed good interactivity with 民可clinical samples in 物嗎clinical trial. In con好可clusion，the prepared m答唱aterials were advantaged by good h鐘時omogeneity，stab知暗ility and high accuracy小動，which had been successfully evalua相店ted by National Administrat分拿ive Committee for CR白謝M's and could be used in the fie是會ld of quality and safety detection of員務 animals and an雨好imal products，so as to ef南小fectively guarantee the preve歌西ntion and contr筆資ol of CSFV.全文下載鏈接：https://kns.cnki.net/學購kcms/detail/detail.aspx?dbcode=玩土CJFD&dbname=CJFDAUT在他O&filename=ZGDW202010022&v=Bh8a8Z海請T44%25mmd2BUcz35雨關7Gp6m%25mmd2FNVNsXMlCDIj%拍亮25mmd2BoMrJ31MCG%25mm的吃d2FRTZxo4dMrPllwgY自廠bd35ap
2020-10-14
PCV4 SYBR Green I實時(shí)熒光定量PCR方姐相法的建立及應用
爲建立一種(zhǒng)靈敏、可快速檢測豬圓環病毒4型（PC年對V4）的SYBR Green I實時(shí)熒光定現費量PCR方法，根據PCV4 Rep基因的保守區域設些雜計引物，建立針對(duì)PCV4的SYBR Gr花自een I實時(shí)熒光定量PCR檢測方法，并對(duì)該方法的特異性外書、靈敏性和重複性進(jìn)行評價。結果顯示，什商該檢測方法的Ct值與标準品在5.64×102~5.學樹64×109 copies/μL範圍内呈良好(hǎo)的線性關公員系，R2爲0.999，斜率爲-3.低煙638，檢測下限爲5.64×102 copies/μL，且無非特異性擴錢水增。利用該方法對(duì)56份臨床樣(yàng)品進(j媽低ìn)行檢測，發(fā)現PCV4核酸陽性率爲3.57%，比普通PCR更靈影鐵敏可靠。結果表明，本試驗建立的SYBR Green I實時理地(shí)熒光定量PCR方法可用于PCV4的快速診斷。Establishme自為nt and Application of SYBR 多雪Green I Real-time PCR for Detection o校喝f PCV4In order to rapidly and 窗麗sensitively detect porc下制ine circovirus ty兵計pe 4（PCV4），the SYBR Green I real-身書time quantitative PCR assay was藍還 established based on the primers東行 that were designe制厭d according to t黑風he conserved region of PCV4呢厭 Rep gene. Then 謝唱its specificity，sensitivity and repeat行技ability were evaluated. T鄉明he results showed that its Ct valu舞睡e remained a good linear relationsh費要ip with the standard materials都笑 within the range of 5.64×10體去9~5.64×102 copies/μL，specif黑筆ically，R2 was 0.999，the slope was -3.63公煙8，and the detection limit was林笑 5.64×102 copies/μL without non-s飛和pecific amplification. 56 clinical sa坐工mples were tested by the method，i費相t was found that the nucleic ac我上id positive rate of P吧看CV4 was 3.57%，which was more sensiti他西ve and reliable女地 than that of conventional PCR兵謝. Therefore，the established SYBR 裡拍Green I real-time PCR method could achi車山eve the rapid detection of P放街CV4.全文下載鏈接：https://kns坐請.cnki.net/kcms/detail/detail.場分aspx?dbcode=CJFD&dbname=店制CJFDAUTO&filename=ZGDW202010021&v=Bh8a8的知ZT44%25mmd2BX%2問好5mmd2B788oXBniPXK%25mmd業能2BKSJj9SS83bukL0TwOF3EqaFRYut6xL7od5eX了雨Fwsk
2020-10-14
血清型特異性水泡性口炎病毒同步檢測和鑒别方法的建立
爲準确地鑒别診斷新澤西型水泡性口炎病毒（VSV-NJ）和厭器印第安納型水泡性口炎病毒（VSV-IND），本光弟研究建立了可以同時(shí)檢測和鑒别診斷VSV-NJ和VSV-IND業請的雙重熒光RT-PCR方法，并對(duì)方法性能(néng)進(城白jìn)行了測試和初步應用。結果顯示，微事建立的雙重熒光RT-PCR特異性爲100%，最低檢舊樂測限1~10個拷貝/μL，擴增效率E值均在99%左右，R2值分别爲0.9開遠99和0.998。VSV-NJ和VS會說V-IND單熒光RT-PCR與雙重熒光RT-PCR的相關性系數分看服别爲0.999 7和0.999 4，與單熒光RT-PCR相比，該引物和探針的老物雙重體系混合對(duì)特異性和敏感性均未産生明顯幹擾。坐懂用已知VSV-NJ及VSV-IND陽性核酸驗證，發(fā)資高現符合率爲100%，對(duì)50份臨床樣(y制雨àng)品的檢測均爲陰性，與單熒光RT新和-PCR結果一緻。本研究建立的雙重熒光RT-PCR方法實現了她司VSV-NJ和VSV-IND的同步檢測和鑒别，爲水泡性口炎的防控提供煙低了技術儲備。Establishme民場nt of a Duplex Real-time RT年妹-PCR for Detection and Identificat區女ion of Serotype-s開也pecific VSV Strai的路nsIn order to accur學吃ately identify and科舞 differentiate the serotype-知藍specific strains of New Jers技票ey type of vesicular st秒山omatitis virus（車你VSV-NJ）and Indiana type of vesicular s飛月tomatitis virus（VSV-IN知媽D），a duplex real-time R飛快T-PCR method was establ為熱ished for simultaneous detection of th雨討e two serotypes of 請醫viruses，and its performa靜媽nce was tested and initial app林農lication was carried ou自個t. The results s人大howed that the s友技pecificity of the established m要長ethod was 100%，with the lo大銀west detection limit of 哥服1~10 copies/μL，both of the amp下哥lification efficien媽冷cies（E）were about 訊遠99%，and R2 values wer開討e 0.999 and 0.998，respectively. The co聽為rrelation coefficients between吃制 singular real-time RT-PCR and du問日plex real-time RT-PCR for VSV-NJ土要 and VSV-IND were 0.999 7 and 0.9土唱99 4，respectively. C很們ompared to sing吧都ular real-time RT-PCR，both the sp坐樹ecificity and sensitivity were 學內not affected by the吃問 mixture of the primers and probes.風南 The coincidence rate wa海電s 100% as verified by the known pos短北itive nucleic aci就嗎ds of VSV-NJ and VSV-IND吃車，and 50 clinical samples were t朋微ested to be negative，which was錯錢 consistent with the results 我舊by singular real-time RT-P謝通CR. As a conclusion，the method establi廠光shed in this study could simu亮說ltaneously detect 的內and differentiate VSV-NJ and VS視妹V-IND，which would請得 provide technical reserves 吃事for prevention and con是章trol of VSV.全文下載鏈接：https鐘湖://kns.cnki.net/kcms亮子/detail/detail.aspx?dbcode=CJFD&dbn我輛ame=CJFDAUTO&filename=ZGDW202010020&窗理v=Bh8a8ZT44%25mmd2BVeM1YDq6a8的得EjAd9sCihEm3ObfSAW0y們站v0S397NvPYV7ZDJgBAp1舞這of7G
2020-10-14
嚴重急性呼吸綜合征冠狀病毒2型4種和輛(zhǒng)結構蛋白特性分析
爲深入了解嚴重急性呼吸綜合征冠狀病毒2型（SARS-Co資算V-2）S、M、E、N蛋白的分子生物學(xu業短é)特征和基本結構特征，選取S、M、E、嗎從N蛋白序列進(jìn)行生物信息事愛學(xué)分析和預測，使用DNAstar、樹劇SignalP、TMHHM、PSORT Prediction、BUSCA和飛請ABCpred軟件，分析并相互驗證4種(zhǒng)蛋白的結構域特征和S蛋白男綠的潛在B細胞抗原表位，結果預測樹熱出了4種(zhǒng)結構蛋白的功能(nén員有g)結構域，并分析預測出S蛋白從著潛在的B細胞抗原表位爲25~29、75~81、112~116能坐、148~152、773~779 aa。研究結果可爲SA土習RS-CoV-2相關的分子生物學(xué)麗歌和結構生物學(xué)研究提供參考，并爲疫苗開(kāi)發(fā)少書等提供理論依據。Characteristics of Four Kinds o雪雪f Structural Pro村大teins of SARS-CoV-2In ord林煙er to further identify the mole個長cular biological characteristics and g媽火eneral structural characteristics of 愛知S，M，E and N proteins of severe ac笑可ute respiratory syndrome coronavirus去地 type 2（SARS-CoV-2），the amino acid sequ關照ences of the above protei見問ns were selected for bioinform西友atics analysis and p志行rediction. The struct鄉行ural domain charact樂生eristics of the above proteins and th廠機e potential B cell epitopes of S prote樂黑in were analyzed and mutuall視理y verified by DNAstar，SignalP，TMH暗些HM，PSORT Prediction，BUSCA and ABC遠喝pred softwares. The re月厭sults showed that the functional dom子地ains of the four proteins were predic中林ted，and the potential B cell epitopes 河木of S protein was analyzed a近歌nd predicted to be 25 to 29，75 to 81慢快，112 to 116，148 to152北年 and 773 to 779 aa.知信 Based on the results，s低司ome references were provided for刀物 further study on molecular biology an內場d structural biology rela筆音ted to SARS-CoV視理-2，and theoretical basis wa爸從s provided for vac吧懂cine development.全文下作畫載鏈接：https://kns.cnki.net/kcms/deta東農il/detail.aspx?dbcode=CJFD&db男影name=CJFDAUTO&filename遠制=ZGDW202010018&v=Bh8a8ZT44%25mmd2BUnxRi們也nWoOlcFhZMdOudDd1JIPIlGMwpAhO習醫sdGh10NuiNy9Bd18v2rS
2020-10-14
犬冠狀病毒研究進(jìn)展
2020年2月，我國(guó)香港報道(dào)了新冠肺員間炎（COVID-19）患者家養的寵物犬感染了新型冠狀病毒（SA懂新RS-CoV-2），這(zhè還吧)引起(qǐ)了人們對(duì)犬冠狀病毒的關注。爲有助照照于人們全面(miàn)了解犬冠狀病毒，做好(h雨店ǎo)犬冠狀病毒病防治，本文從病原學(xué)與流行病學(x刀制ué)、臨床症狀以及診斷和防治措施等方面(miàn)進(jìn)行了樹吧綜述。犬冠狀病毒在犬群中廣泛存在，主要包括α冠狀病毒屬的犬腸炎冠狀病毒也地和β冠狀病毒屬的犬呼吸道(dào)紙對冠狀病毒。犬冠狀病毒緻病性不強，引起(qǐ)的臨床村光症狀通常較輕，但對(duì)幼犬或與犬細小病毒、犬瘟熱病毒等發(fā費廠)生混合感染時(shí)，會(huì)引起(qǐ)較嚴重症技村狀，且易發(fā)生突變，産生影坐新的變異毒株，具有緻病性增強的風險。目前，林黑犬冠狀病毒檢測方法主要包括病原學(x和資ué)檢測、血清學(xué)檢測以及RT-PCR、qRT-PCR、納米PCR等姐去分子生物學(xué)檢測。犬冠狀病毒感染的防不水治主要通過(guò)疫苗免疫以及科綠會學(xué)飼養管理、衛生消毒和對(duì湖場)症治療等綜合措施。雖然犬可感染SARS-CoV-2，但尚不清楚感染裡分犬是否可將(jiāng)病毒再傳染給其他動物或空短人類。因此，應加強犬冠狀病毒的病原生态學好我(xué)研究和監測預警。Rese黑去arch Progress on Canine CoronavirusIn 理工February 2020，i拿個t was reported in Hon黃如gkong，China，that a pet dog kept by a用山 patient confirmed with c也廠oronavirus disease 2019（C黃微OVID-19）was infected 司草with severe acute respiratory syndrome 我綠coronavirus 2（SARS-船車CoV-2），which caused the concern about答爸 canine coronavirus. In order to f弟從ully understand the study on canine cor樹山onavirus，prevent and control這知 the disease，the etiology and epidemi資子ology，clinical symptoms，meas吃年ures of diagnosis and con船年trol were summarized in the paper. Th個知e canine coronavirus was widely distrib動熱uted in dog populations，ma報睡inly including canin對理e enteritis coronavirus of Alphacor事歌onavirus and canine respi在舞ratory coronavirus of Beta去費coronavirus. The canine coro道靜navirus was with poo書鐵r pathogenicity，which could cause slig區算htly clinical sym個愛ptoms，but would be more ser作議ious and likely to mutate to new mutan高子t strains when mixed infect嗎市ion of it with c他習anine parvovirus and canin站見e distemper virus oc醫東curred，with the risk唱民 of increasing pathogenicity. A水道t present，the detection m關民ethods for canine 們林coronavirus wer厭知e mainly including pathogen d市是etection，serological detection，能聽and molecular biolog不的ical detection methods suc南了h as RT-PCR，qRT-PCR and N河玩ano PCR. Canine coronavirus co化跳uld be controlled by a ser店短ies of comprehensive measur兒技es including vaccina美鐘tion，scientific feeding and身子 management，disinfection and sympto照煙matic treatment，etc答花. It was still unclear 理件whether the virus could be sp愛白read to other animals or human小短 via the infected dogs暗鐘，although dogs could 民分be infected with SARS林鐵-CoV-2. Therefore，it雜美 was necessary to strengthen st請城udy on pathogenic eco畫了logy as well as monitoring and early近區 warning against the viru我兒s.全文下載鏈接：https://kns.cnki.ne體業t/kcms/detail/detail.aspx?dbco的電de=CJFD&dbname=CJFDAUTO&filename=ZGD懂劇W202010017&v=Bh8a8ZT44%25mmd2BXJIZxJ窗嗎9AXLnmQLd%25mmd2BGaTsBTZkV2VNkT6%25mmd2外電By5cE%25mmd2F1YOsAS0綠民uFHMQ4f5Dz
2020-10-14
國(guó)内外奶牛疫病預警監測技術發(fā)展現狀
爲了解國(guó)内外奶牛疫病預警技術街也及其應用現狀，歸納了奶牛疾病的預警要求，分析了國(guó)内外8種說鐵(zhǒng)疫病預警技術的發(fā)展現狀及其購輛在奶牛養殖中存在的局限性。一是獸醫巡檢預警技術：該的海技術較爲成(chéng)熟，但不能(né機現ng)滿足大規模養殖牛場迅速、高效預警的需求。二是奶樣(yàn舊間g)體細胞數測定技術：可初步診斷隐性乳腺炎，但需要事習額外細菌培養和檢測确診。三是奶牛群體改良測定技術：以監測生産性能(néng房拿)爲主，僅可預警奶牛乳腺炎及酮病。四是計步器及發(文器fā)情、反刍項圈預警技術：同時(shí)具備發(fā)情與反刍監近又測功能(néng)，但僅适用于牛群的發(fā)情與反刍監測。五是大數據挖掘亮聽技術：在數據庫基礎上，利用各種(zhǒng)模型或算法預測傳染病的發(fā)生時舊和發(fā)展，但預警效率受限。六是智能(néng)化體溫連續遠程監測及預警技但亮術：可預警口蹄疫等重大傳染病，但無法完全替代臨床診斷，不能年雜(néng)監測病原微生物。七是區紙基于AI和群體熱成(chéng)像的奶牛行爲分析技術：可以監視記錄奶服了牛行爲與溫度特征，鎖定病畜，但對(duì)發(fā)病初期無異綠大常行爲的牛隻無效。八是生物氣溶膠激光光譜測量技術：通過(guò)檢測病票快原微生物種(zhǒng)類和濃度，達到預警疫病的目的，目前主聽對要用于軍方監測人類重大疫病。未來的奶牛疫病預警技術需要進(jì睡長n)一步相互結合、取長(cháng)補短，充分利用大數據、物聯網等新興技家音術進(jìn)行奶牛重大疫病預警診斷。本文爲預警監測明說技術的本地化發(fā)展提供了參考。 Develop相睡ment Status of Early Warning 間河and Surveillance 都到Technologies for C多森ow Diseases In order to id可農entify the development and applicati站又on of early warning在銀 and surveillance technologies 商在for cow diseases in the world，relev是離ant requirements for the te紅個chnologies were summarized，and the dev可村elopment status of 8 different te說老chnologies and their 章弟limitations in cow 紅老breeding industry were analyzed.年女 Specifically，for the tec校畫hnology of routing inspection 做快and early warni微請ng by veterinarian，歌費it was relatively mat公水ure，but failed to 科音meet the need for謝下 rapid and efficient early 紅體warning in large-scale farms；for the老做 technology of somat子事ic cell count（SCC）of milk sa筆船mples，by which，recessive masti生要tis could be initially diagnosed，but得紅 additional bacteri船是al culture and testing woul美慢d still be needed；for the dairy he子體rd improvement（DHI）technolog自有y，it mainly focused on the來土 monitoring of produ場內ction performance，and onl喝日y could be applied to early wa高我rning of cow mas間放titis and ketosis；for the pedometer a科日nd collar early war跳黑ning technology for estro窗問us and ruminant，it was only appropri爸笑ate for monitoring of estrous and 雜離ruminant of the cows；for big dat機坐a mining technology，t這師he occurrence and development o山去f infectious diseases could be 女業predicted by various m冷一odels or algorith去東ms based on the database，but its刀森 efficiency was limited；for intellige師紅nt continuous remote tem長間perature monito事不ring and early warning姐件 technology，major infectious diseas到友es such as foot-and-房可mouth disease could be war西習ned，but it could not fully replace clin木短ical diagnosis and failed to 遠是monitor pathogeni購理c microorganisms；for th知新e cow behavior anal放說ysis technology based 低爸on AI and population thermal ima劇制ging，the behavior and temp商服erature characteristics of腦計 cows could be 電老monitored and recorde微間d，sick cows could b但店e targeted，but the technology was inv歌報alid for the cows witho大了ut any abnormal behavior場來s at the early stage of the街站 disease；and for bio-aerosol laser場場 spectrometric measurement tec還藍hnology，the purpose慢用 of early warni友快ng could be achieved through th月大e testing of species and concentration 月個of pathogenic microorganism做村，which was mainly used to 森遠monitor major d師湖iseases in human by milita照頻ry at present. In c愛兒onclusion，the early warning te我錯chnologies for cow diseases sho看民uld be further c爸師ombined with each other in the fu動家ture，learn from each oth件房er and make full use of big dat去電a，internet of things and other emer北姐ging technologies，水林which would contribute to early warn可上ing and diagnosis of major cow disea黑能ses. Some references were provi短用ded for the localization developme知短nt of early warning and surveillance te數年chnologies. 全文下載鏈接：ht錢自tps://kns.cnki.net/KCMS/deta化路il/detail.aspx?dbcode=CJF舊話Q&dbname=CJFDAUTO&filename=ZGDW202010購近016&v=MjUzODc0SE5ITnI0OUVZb1I身子4ZVgxTHV4WVM3RGgxVD農村NxVHJXTTFGckNVUjdxZVorZH工坐NGeS9rVTc3SlB5clBl相文Ykc=
2020-10-14
我國(guó)部分地區送檢樣(亮老yàng)品3種(zhǒng)常見豬病血清學(xué)和病原吧機學(xué)檢測
爲了解我國(guó)部分地區豬瘟、豬繁殖與呼吸綜合媽綠征和豬僞狂犬病3種(zhǒng)常見疫病的流行情況，采用EL快科ISA、聚合酶鏈式反應（PCR拍報）和反轉錄PCR（RT-PCR）等方法，對(duì)2019年1報草1個省（自治區）172個豬場送檢的3 550份豬血清樣(yàng)品進又農(jìn)行豬瘟病毒（CSFV）、快樂豬繁殖與呼吸綜合征病毒（PRRSV）及僞狂犬病野毒（PRV-gE）抗體檢測，劇路對(duì)9個省（自治區）72個豬場送檢的371份厭習病料進(jìn)行抗原檢測。結果顯示：CSFV、PRRSV、PRV姐就-gE抗體陽性率分别爲82.75%、84.08%、31.動紙86%，抗原陽性率分别爲2.62%答月、26.15%、1.67%。保育豬群CSFV抗體平均阻斷學對率最低，爲44.62%，抗體整齊度差，CV爲58.2嗎動6%，且CSFV抗原主要在哺乳仔豬和保育豬病事數料中被(bèi)檢出，檢出率分票西别爲3.57%、3.00%；除育肥豬群外，其站北他各階段豬群的PRRSV抗體S/P平均值爲1.0~2.0能爸，抗體整齊度整體較差，且平均抗長照原陽性率較高（26.15%），其中保育階段豬群病原檢出率場商最高（37.68%）；育肥豬、母豬、公豬的P低廠RV-gE抗體陽性率分别爲36.82%、35.31%、15.65%，病原主要從能來死胎/木乃伊胎和哺乳仔豬病料中被(bèi)檢出，檢出率分别爲8.70%和2.藍長99%。結果表明，PRRSV對(duì)我國(guó)豬場不同階段豬群均微吃有較大威脅；保育豬群CSFV抗體水平不理想，暴發(fā)疫情可長風險較高；公豬、母豬及育肥豬群普遍存在PRV感染，尤其是母豬。提議看土各豬場采取免疫、監測、淨化等綜合措施，加請畫強這(zhè)3類常見豬群疫病的防控。Serological and Path年離ogenic Detection of Three Commo白身n Swine Diseases照都 in Some Region資下s of ChinaIn order to iden煙匠tify the prevalence status of three c說得ommon swine diseases including cl雨讀assical swine fever區分（CSF），porcine r船懂eproductive and respiratory syndrome（P鐵票RRS）and porcine pseudorabies（PR）in算門 some regions of China，by the me說熱thods of ELISA，PCR and R說南T-PCR，3 550 swine se的嗎rum samples delivere動舞d by 172 farms in 11 provinces（a信農utonomous regions）in 2019短討 were tested for the ant那水ibodies against CSFV，PRRSV and PRV-g短志E，and 371 lesion samples from 72 f看知arms in 9 provinces（autonomo醫購us regions）were detected 舞化for virus antigens. The results 光短showed that the antib妹愛ody positive rates of CS民下FV，PRRSV and PRV-gE we唱理re 82.75%，84.08% and 31.86% r睡還espectively，and the anti們拍gen positive rates山放 were 2.62%，26.15% 飛腦and 1.67% respectively. 來黃The average blocking rate火喝 of CSFV antibody in nursery pigs 費空was 44.62%，which was the low子人est with poor antibody uniform煙司ity，and the CV value w爸白as 58.26%，CSFV wa亮道s mainly detected from the相體 lesions of suckling p議可iglets and nursery pigs為讀，with the detection rates of 3.57% an木唱d 3.00% respectively. E的從xcept for fattening pi兵飛gs，the mean S/P value of P媽一RRSV antibody was 1.0 to 2.0 for other志她 pigs，with poor antibod工時y uniformity，and t讀日he average positive rate of PRRSV a雜文ntigen was higher（26.15%），in 刀森which，the detection rate 業麗of PRRSV antigen i票路n nursery pigs was the highest（37.就們68%）. The positive rates o微土f PRV-gE antibody in fat做資tening pigs，sows an舊子d boars were 36.82%，35.31% and 15秒雪.65% respectively，and the virus pathoge場是ns were mainly detect些熱ed out in stillborn/mummy這去 fetuses and lesions of suckling光白 piglets，with the positive r到兒ates of 8.70% and 2.9術雨9% respectively. It was concl笑人uded that PRRSV had brought great 的們threat to pig pop喝讀ulations at different gr見朋owth stages in China對少；the nursery pigs an人些tibody level of CSFV地文 was poor with hig水裡h risk of outbrea購拿k；boars，sows and fatte電有ning pigs had been 村討widely infected with PRV，especially so行說ws. Therefore，a series of compreh鄉章ensive measures including immunizat訊票ion，monitoring and purificati這商on and so on should be applied in all f女船arms，so as to strengthen the 跳南prevention and control of thes鐘農e common diseases. 全文下載鏈接：https://kn拍電s.cnki.net/kcms/detail/detail.aspx?機友dbcode=CJFD&dbn哥的ame=CJFDAUTO&filename=ZGDW2020100謝海05&v=Bh8a8ZT44%25m司件md2BXxmHa%25mmd2腦相F3pXZzppSm8lNN5tOt上湖vxTeoAZGuhyS9TRPv%25mmd2FYabVKxDG%2亮外5mmd2Bf1nv
2020-10-14
雲南省邊境地區送檢牛血清樣(yàng)品藍舌麗黃病病毒抗體檢測與血清型鑒定
爲了解雲南邊境地區的藍舌病流行和分布情況，采用競争ELISA亮道方法，2019年對(duì)雲南省邊境地區5個州市12個縣（市）的1 5在湖80份牛血清樣(yàng)品進(jìn)行藍舌病病毒錯們抗體檢測，同時(shí)從檢出的陽性血清中，每縣（市）随機選取15~55份，采費生用微量血清中和試驗進(jìn)行血清型鑒定。結果顯示：12多弟個邊境縣（市）均檢出陽性血清樣(yàng)品，但有一定的地區兒一差異，其中芒市最高（94.1%），滄源縣最慢費低（17.0%）；本地牛和境外牛血清樣(yà秒靜ng)品陽性率差異不大（P＞0.05），均在70.0%以上；各縣（鐘得市）均存在5個以上血清型的交叉感染。結果表明，藍舌病在雲南姐亮邊境地區廣泛存在，且呈多種(zhǒng)血清型混合流行态勢，需要持續女員開(kāi)展血清學(xué)調查。本研究爲銀看我國(guó)西南邊境地區藍舌病防控提供了數據參考。Detection 他務of Bluetongue Virus Antibodies in Bovi空麗ne Serum Samples from Border Are訊離as of Yunnan Province a我黑nd Relevant Serotype Ide綠體ntificationIn order討學 to investigate the prevalence and術知 distribution of blue有紙tongue disease in border areas of Y東為unnan Province，1 580 cattle serum samp行市les collected from 12為化 counties（cities）of 5公門 prefectures（municipalities）in border睡從 areas in 2019 were t藍視ested for blueto少個ngue virus antibod笑友ies. For the positive空厭 serum samples，15 to 55 samples were r媽工andomly selected from each co可小unty（city）to identi友好fy serotypes by micro-很爸serum neutralization站吧 test. The resu要關lts showed that positive serum sam通報ples were detected ou們喝t from all the 12 border countie新紅s（cities），with certai家件n regional differe機亮nce，among which，maxi視花mum positive rate occurred in Mang Cit技雪y（94.1%）and minimum rat畫現e in Cangyuan Count城電y（17.0%）. The serum posi都他tive rate of local cattle was slig業紙htly different from tha男數t of foreign cattle技資（P＞0.05），both o低器f them were higher than 70窗錢.0%. Mixed infect厭制ion of more than 5 seroty作湖pes was present in al月你l counties（cities）. It was con女理cluded that bluetongue disease was站冷 widely distributed in the border ar數數eas，and tended to spread 你嗎with mixed serotypes，so serolo分小gical investigation 中間should be continued to carry out到分. Some data references were provi都個ded for the prevention and control遠個 of the disease in the border areas城短 in Yunnan Province.全文下載鏈接：ht妹這tps://kns.cnki.net/kcms/detai河制l/detail.aspx?dbcode=CJFD&dbname=C熱快JFDAUTO&filename=ZGDW20201訊媽0004&v=Bh8a8ZT44%這農25mmd2BUA18IL%25mmd2FvvYlJeua9SVCqZZW年書mUVoNbW7uYr4JfTu5x3Fjl5GE%25mmd2FC6校報i6H
2020-10-14
豬瘟病毒和牛病毒性腹瀉病毒雙重熒光RT匠低-PCR檢測方法的建立
爲同時(shí)檢測豬源臨床樣(yàng)本中的豬瘟病毒（CSFV）北吧和牛病毒性腹瀉病毒（BVDV），基于這(zhè)兩(liǎng)種(知山zhǒng)病毒的5'UTR序列設計特異引物和多車TaqMan探針，建立了一種(zhǒng)檢測CSFV志章和BVDV的雙重熒光RT-PCR檢測方能科法，并對(duì)該方法的特異性我河、最低檢出限和重複性等進(jìn)行了評價開歌。結果顯示，該方法隻對(duì)CSFV和BV道開DV呈現特異性擴增，對(duì)豬僞狂犬病病毒、豬爸和繁殖與呼吸綜合征病毒、豬傳染性胃腸炎病毒、豬流行性腹瀉病毒、豬圓環病毒2嗎唱型不發(fā)生交叉反應，對(duì)陽性标準對(duì)照CSFV-5村店'UTR-RNA、BVDV-1-5'北頻;UTR-RNA和BVDV-2-5'UTR-RNA，金用最低可分别檢出27、36和32拷貝/空西μL。該方法的組内和組間試驗Ct值變異系數介于0.11人區%~1.20%，具有良好(hǎo)的重現性家紅。對(duì)152份豬組織樣(yàng)本用白數該方法進(jìn)行CSFV和BVDV核酸檢測，結果山我檢出CSFV陽性樣(yàng)本16份，BVDV陽性樣(yàng)本3年月份，CSFV和BVDV雙陽性樣(yàng)本1事這份，與國(guó)标和OIE《陸生動物診斷試驗和疫苗》相應的熒劇跳光RT-PCR方法陽性符合率爲100%。結果表明，本研究建立的雙重熒光RT-P可銀CR方法可用于臨床樣(yàng)本中的C黑那SFV和BVDV檢測，從而爲豬瘟防制和朋藍淨化提供了一種(zhǒng)有效的技術手段。Establis銀自hment of a Duplex Fluorescen分腦t RT-PCR Assay for D雪算etection of Cla木林ssical Swine Fever Virus and B熱妹ovine Viral Diarrhea VirusIn o可秒rder to simultaneously 為體detect classical男白 swine fever virus（CSFV）and bovine vir門高al diarrhea virus（BVDV長車）in clinical samples新嗎 from pigs，the specific primers a兒外nd TaqMan probes wer為街e designed based on 5'UTR 厭些sequence of the two kinds of viru靜議ses，and a duplex fluorescent RT-P少校CR was established，the specif藍銀icity，minimum detection limit and repea房謝tability were evaluate理劇d. The results 看聽showed that the assay could reac大又t specifically with CSFV歌睡 and BVDV only，的樂but failed to crossly身視 react with other viruses including 海友pseudorabies viru他動s（PRV），porcine reproductive and resp校業iratory syndrome virus（PRRSV）黃能，transmissible gastroenteritis viru我事s（TGEV），porcine epidemi器文c diarrhea virus（PEDV）and porci小舊ne circovirus-2（PCV-2）. 村我The minimum detec身懂tion limits were 27，36 and 32 copie湖木s/μL for the positive standard是現 plasmid control of CSFV-5'UTR音站-RNA，BVDV-1-5校微9;UTR-RNA and BVDV-2-5'UTR-RNA新請 respectively. The月相 coefficients of variation（CV）綠媽of intra-and inter-group ranged 0.11% t見朋o 1.20%，showing good re空新producibility. Atotal of 152 tis南多sue samples were tested for CSFV and子窗 BVDV by the assay，with the results of 知呢16 CSFV positive samples，3 BVD爸事V positive samples an著銀d 1 CSFV-BVDV dual pos外務itive samples，which were completely con店子sistent with the nat西對ional standard a農多nd the results of corr短著esponding fluore通河scent RT-PCR as這靜say specified in OIE 現新Manual of Diagnostic Te工醫sts and Vaccines for Terrestrial Anima明雨ls. In conclusion，the assay establis什那hed in this study could b計山e used for the detec樹喝tion of CSFV and BV拿文DV in clinical samples，which provided a放輛n effective techn計線ical method for control a離畫nd purification of the two dis下暗eases.全文下載鏈接：https://kns.c志這nki.net/kcms/det街雜ail/detail.aspx?dbcode=CJFD&dbname=CJ微湖FDAUTO&filename=ZGDW202009022&v=B笑雜h8a8ZT44%25mmd2BXfswhdu到厭qZcZQVHMHhHXi9wSi2xg6rwQX6OsZDt動車jLz8LFamaifdRAHg
2020-09-27
鹦鹉熱衣原體熒光環介導等溫擴增（LAMP）檢測方法的建立
鹦鹉熱衣原體（Chlamydia psittac我森i）是一種(zhǒng)人獸共患病原體，能(néng)引起(我雨qǐ)自然疫源性衣原體病，目前廣泛分布于世界各地。本研究應用環介導等溫擴增技妹窗術（LAMP），針對(duì)鹦鹉熱衣原體23S物煙 RNA基因序列設計引物，并進(jìn)行篩選以及敏感車近性和特異性試驗，建立了鹦鹉熱衣原體恒溫熒光擴增要技方法。該方法在63 ℃恒溫條件下1 h内即可顯示結果，操作簡單土物、快速，特異性強，靈敏度可達100拷貝/µL，且與流産衣問開原體、動物布魯氏菌、牛結核分枝杆菌、沙門氏菌、大腸杆海哥菌、金黃色葡萄球菌無交叉反應；用光很國(guó)産儀器可直接通過(guò)檢測熒光信号判讀結果，既增強了看是準确性，也避免了開(kāi)蓋污染産生假陽性；應用員哥建立的LAMP方法對(duì)實驗室保存的臨床DNA樣(yàn頻熱g)品進(jìn)行檢測，發(fā)現檢測結果與QPCR相同。該方法的建立彌補兵和了傳統檢測技術的不足，爲實現鹦鹉熱衣原體的現場快速診斷提供了技術支持事草。Establishment of a Loop-mediated Isot讀場hermal Amplification Method說話 for Chlamydia psitta樹藍ciChlamydia psittaci，a 動暗kind of zoonotic pathogen，is widel也男y distributed all over店國 the world，by which開空，chlamydiosis of natural origin mig長動ht be caused. In this study，the prim技畫ers were designed bas間算ed on 23S RNA gene sequence和刀 of Chlamydia psittaci，and then路答 screened. After sensitivity and sp路朋ecificity tests木鐵，a thermostatic fluorescence ampl有舞ification method for Chlamydia psi地體ttaci was established according to the技有 loop-mediated isothermal amplification老黑 method（LAMP）. By the e用公stablished method街舊，test results could be shown within one暗匠 hour under the consta對友nt temperature of友老 63 ℃ due to its advantages i你算ncluding simple operat信銀ion，rapidity，strong spe長不cificity，sensitivity of 100 co著服pies/µL and no cross-reac書暗tion with Chlamydia 秒要abortus，Brucella an司頻imalis，Mycobacteri河到um bovis，Salmonella，Escherichia col鐘明i and Staphylococcus aureus. T年外he results could b話購e interpreted directly by fluore是線scence signals with domestic instr件去uments，which not only enhanced資知 the accuracy，but also avoided fals那來e positive due to pollution brought離姐 by cap opening. The clinical D雜說NA samples stored in laboratories were 窗哥detected by the LA票很MP method，it was found th民也at the results were same as th會來at by QPCR. It was concluded tha呢視t traditional m化暗ethods were remedied by the establ人動ished method，which provided techn亮跳ical support for rapid f說我ield detection of Chla筆女mydia psittaci.全文下載鏈接：https://kns.cn草少ki.net/kcms/detail/detail.aspx?dbcode=學工CJFD&dbname=CJFDAUTO&filename=ZGDW20200綠但9021&v=Bh8a8ZT44%25mmd2BXxG什厭njxzXiJU3p%25mmd2Fivz暗水uIEWb0bnO6OLe8PeO%25mmd2Bm音自d6CaSRqBjEGndhokrs
2020-09-27
牛結節性皮膚病診斷方法研究進(jìn)展
牛結節性皮膚病（lumpy skin disease，LSD）得見是由牛結節性皮膚病病毒（lumpy skin dis物時ease virus，LSDV）引起(qǐ)的急性、亞光銀急性傳染病。我國(guó)于2019 年8 月在新疆伊音媽犁地區首次确診該病。快速準确檢測是防控該病的關鍵。目前，應用常規光和PCR、實時(shí)熒光定量PCR、高和他分辨率熔解曲線分析、環介導等溫擴增等分子檢測方法，已實現了紙雪LSDV與其同屬不同種(zhǒng體制)病毒的通用檢測和鑒别檢測以及LSDV野毒株與疫苗株的區分；應用女麗病毒中和試驗、酶聯免疫吸附（ELISA）和蛋白印迹試驗（WB）等免疫學(xué車校)方法，可進(jìn)行羊痘病毒屬的特異志煙性抗原與抗體檢測，而對(duì)于LSDV特異性抗體檢測方校站法尚有待研究。LSD作爲一種(zhǒng)新傳入我國(guó)的腦吃外來動物疫病，有必要運用快速準确的分子或免疫學(x腦個ué)檢測方法加強監測。本文概述了LSD診斷方法研究進(jìn)展，愛問爲我國(guó)LSD診斷技術研發(fā)提供參考。 Res了西earch Progress on Diagnostic 西少Methods of Lumpy Skin DiseaseLum家光py skin disease（LSD），an acute an麗樂d subacute infectious disease，購工is caused by lumpy skin disease vi嗎放rus（LSDV）. In August 2019，Chinese firs文雜t case was confirmed in 有理Yili Prefecture，Xinjiang. To pr明為event and control the di訊麗sease is subject to rapi算討d and accurate detection. Presentl雜電y，the goal of generic d生花etection of LSDV and its congeneric門師 virus of different spec道劇ies as well as differential雜畫 diagnosis of wild科車 strains and vaccine strains have been 懂和achieved by a serie物腦s of molecular detection method從湖s，such as conventi數們onal PCR，real-time f司文luorescent quantitativ動火e PCR，high-resolution melting（HRM）cur農雪ve analysis，loop 用河mediated isothermal amplificatio爸線n（LAMP）；the specific antigen and antib謝請ody of capripoxvirus could b下媽e detected by the immunologica熱女l methods，such as virus neut空動ralization test（VNT），enz家拍yme-linked immunosorbent assay（ELISA），W機些estern Blot（WB），etc. However，the det些裡ection methods for speci笑西fic antibodies 我煙against LSDV still need further st電和udies. As a new年舞 exotic animal diseas湖和e，LSD should be furthe裡用r monitored by rapid and accurate molec冷但ular or immunological methods. In c黑我onclusion，relevant research progress門個 was summarized for the purpose 有區of providing some references電風 for the development of LSD dia理爸gnosis technology i嗎北n China. 全文下載鏈接：https://讀路kns.cnki.net/kcms/detail/detail.林要aspx?dbcode=CJFD&dbname=CJFDAU銀草TO&filename=ZGDW202009018&v=Bh8a話影8ZT44%25mmd2BWJa低空zKXHZJ2lAttfvyxPfcF6AGOiS28OYFQ理年zAbEmEhkOi8ryjg0p66%25mmd2B
2020-09-27
副豬嗜血杆菌的實驗室檢測技術
冠狀病毒在自然界中普遍存在，可感染多種(zhǒn照國g)哺乳動物和鳥類，給畜牧業生産安全和副豬嗜農藍血杆菌病發(fā)病率和死亡率較高，給養豬業造成(開要chéng)了嚴重損失。爲及時(shí)确診該病，本文從細菌分離鑒雨中定、血清學(xué)檢測和分子生物學了河(xué)檢測3個方面(miàn)，對(duì遠信)副豬嗜血杆菌實驗室診斷方法的研究西如進(jìn)展及其優缺點進(jìn)行了綜述。細菌分離鑒定特異性較強，作如但操作複雜、用時(shí)長(cháng)、內爸所需儀器多，無法滿足快速檢測需求；酶聯北做免疫吸附試驗（ELISA）、間接血凝試驗（IHA）和補體鄉市結合試驗（CF）3種(zhǒng)血清學(自個xué)檢測方法操作簡便、快速，但特異性和敏感性不高，主要适用于早期山雨診斷和篩查。近年來，分子生物技術快速發(fā)展，已建立信那了PCR、熒光定量PCR、數字PCR和環介導等溫擴增（LAMP）等多種草快(zhǒng)分子生物學(xué)城鐵檢測方法。這(zhè)些分子生物學(xué)檢測方法敏感、快速、特異和穩司呢定，其中PCR、熒光定量PCR已廣泛應用于實道唱踐中，而數字PCR和LAMP檢測技術尚需進(路知jìn)一步優化。多種(zhǒng)特異性好(作吧hǎo)、靈敏度高、重複性好(hǎo)、檢測快速的實驗室檢測我姐方法的建立和改進(jìn)，爲副豬嗜習道血杆菌病檢測和流行病學(xué)調查提供了有效的技術支撐。 L地裡aboratory Testing Technologies for Hae麗會mophilus parasui綠著sHaemophilus parasuis has brought hug自知e losses to pig industry due to it習慢s high morbidity and mortality. 說匠In order to confirm the dise務司ase timely，the research p城年rogress on relevant laboratory te錢少sting technologie唱船s as well as their advantages and di匠黑sadvantages were summarized from日著 the aspects of isolation and id村好entification of bacteria，serologic線是al detection and molecular biological人慢 detection. Specifically，the method of 亮歌isolation and identificati刀從on of bacteria was with strong specific開吧ity，but was complex and time-為車consuming for ope下一ration，which would req不藍uire for more inst照事ruments and fail to mee雜飛t the needs of rapid detect暗森ion；the serological methods including和資 enzyme linked immunosorbent assay（ELIS門美A），indirect hemagglutination assay（IHA黑電）and complement fixation test件知（CF）were simple動中 and rapid，mainly appropriate for業要 early diagnosis 來黃and screening，but their specif你我icity and sensitivity were relati購是vely low. In recent技呢 years，with rapid development of molecu筆但lar biotechnology，a variety 雜地of molecular biological methods includi快和ng PCR，fluorescence quant謝南itative PCR，digital P湖討CR and loop mediated isother影鄉mal amplification（LAMP）離城had been established，which were師山 sensitive，rapid，specific and st討外able，especially，PCR and fluorescen現海t quantitative PCR 好著had been widely used in 看會practice. But digi腦購tal PCR and LAMP s個嗎till needed to be further會熱 optimized. In concl書好usion，an effective technical support w鄉姐as provided for de東有tection of the disease and e她來pidemiological investigatio房工n through establishing and imp新上roving a variety of laboratory testing 物頻methods with the advantag的雨es of good specificity and rep森就eatability，high sensitivi行哥ty and rapid detection sp為間eed. 全文下載鏈接：https://kns.cnki.net/kcms/d南草etail/detail.aspx?dbcode=CJF對作D&dbname=CJFDAUTO紙道&filename=ZGDW202009017&v=Bh8a8ZT44%25動司mmd2BW9paznIkMtVg舊匠Yb7DdBga1vvYUGvtvTHRfdlrLjxzvQYt%25mmd2嗎快B6li8ZxOQy
2020-09-27
我國(guó)不同地區豬源緻瀉性大腸杆菌的多重PCR報工鑒定及2b-RAD序列分析
爲了解我國(guó)不同地區屠宰生豬肉品中緻瀉性大腸杆菌污染情書分況及系統進(jìn)化關系，降黃計低由其帶來的公共衛生危害，將(jiāng)近一紙幾年分離自華北、華東、華中及西南地區豬屠體表面(miàn)的283株大腸杆菌美樹進(jìn)行了緻瀉性大腸杆菌多重PCR鑒定及2b-RAD測序分型。多月是重PCR鑒定結果顯示，共檢出36株緻瀉性大腸杆菌道些，檢出率爲12.7%，其中西南地區檢出率（26.92%）腦資高于華北（13.33%）、華東（10.57%）和華中（10.81%）民國地區；5種(zhǒng)緻瀉類型中，産腸毒素大腸杆菌（ETEC）、腸緻瀉性大腸朋舊杆菌（EPEC）、腸聚集性大腸杆菌（E問短AEC）、腸出血性大腸杆菌（EHEC）和做畫腸侵襲性大腸杆菌（EIEC）的占比分别爲33.33%、弟男30.56%、25.00%、5.56%和在書5.56%。將(jiāng)緻瀉性大腸杆靜花菌進(jìn)行2b-RAD測序，種(zh學友ǒng)群聚類分析顯示，這(zhè)36株緻瀉學拍菌可分爲8個亞群；系統發(fā)生樹分析顯購美示，親緣關系較近菌株的緻瀉類型基本一緻，分請體離時(shí)間也比較接近，而且來自同一地區菌株的低對親緣關系相對(duì)較近。可見紙問，我國(guó)屠宰生豬肉品中污銀地染的緻瀉性大腸杆菌以ETEC、EPEC和湖美EAEC類型爲主，其流行分布有一定的地域差異，且其親緣關系呈水現一定的時(shí)空特異性。本研究爲精準防控生豬屠宰環節緻瀉性大腸杆菌污信費染，保障肉品衛生安全提供了科學(xué)數據和技術支撐。 Multipl書讀ex PCR Identification 明書and 2b-RAD Sequence Analysis of金通 Porcine Diarrheage船腦nic Escherichia coli i吃裡n Different Regions in Ch照愛inaIn order to make clear the s厭中tatus of contamin訊們ation and phylogenetic relationship長朋 of diarrheagenic Escherichia會短 coli（E.coli）in raw 森老pork products in different regio錢月ns of China and to reduce 音師the public health hazards t山山hat might occur therefrom，diarrheagenic東些 E.coli strains were iden到個tified by multiplex PCR from 283 E.col現玩i strains isolated f相子rom the surface of pig carcasses i還新n North China，East Chin呢票a，Central China and Southwest China i站廠n recent years，then 2b-RA廠美D sequencing and gene typing were car老明ried out. It was found that草吃，by multiplex PCR，36 strains o訊還f diarrheagenic E.coli were de影她tected out，with the det錢學ection rate of 12.7%，among whic開呢h，the detection rate in South自校west China（26.92%）was signifi城家cantly higher than those in North Chi音近na（13.33%），East China（10.57%）and Centr來睡al China（10.81%）；for the f慢但ive types of diarrhea，enterotox們她igenic E.coli（ETEC），e你為nteropathogenic E.coli（樂志EPEC），enteroaggregati業舊ve E.coli（EAEC），enterohemorrhagic E.暗知coli（EHEC）and enteroi物朋nvasive E.coli（EIEC）accoun電城ted for 33.33%，30.56%，2司街5.00%，5.56% and 5.56%，respectively.冷多 After 2b-RAD sequenci畫亮ng for species clustering校了 analysis，it was found t間錯hat the 36 strains could be classifi少訊ed into 8 subgroups；舞請it was shown that，她高by phylogenetic鐘醫 tree analysis，the strains wi金師th close genetic relationship belong不件ed to similar diarrheagenic types，and t睡小heir isolation time was also cl拍地ose，in particular，the str志的ains from the same region w多唱ere with closely呢說 genetic relations人動hip. It was concluded 為劇that the diarrheagenic E.co信自li contaminated in raw pork products哥林 mainly included three types of 火照ETEC，EPEC and EAEC，林物the diarrheagenic E.coli strains were器視 distributed with regional diver現吃sity，and their geneti哥校c relationships were with ce間能rtain spatial and temporal specifici吧路ty. In conclusion，some scientific d農務ata and technical support w老話ere provided to女也 prevent and contro歌街l the containment with 很是diarrheagenic E.coli during the pro鐵通cess of pig slaughtering and to 一紙safeguard hygiene and safety of pork pr區校oducts.全文下載鏈接：https://kns.cnki.net/kc作你ms/detail/detail.aspx?dbc兵明ode=CJFD&dbname=CJFDAUTO&fil作分ename=ZGDW20200得務9010&v=Bh8a8ZT44%25mmd2BUeRq5k%25mm家不d2BkfhEPYt2%25mmd2FmPx9quStu地白%25mmd2F7HhxzlR4Jr8q妹請SYJnW%25mmd2B7QKhWBaK車長Bd
2020-09-27
基于實時(shí)熒光PCR法定量檢測食品中雞源笑一性成(chéng)分
肉類食品真僞鑒定是目前食品安全檢測領域的重點研究内容之讀中一。爲準确定量檢測食品中的雞源性藍了成(chéng)分，根據現行檢驗檢疫标準來南（SN/T 2727—2010）合成(chéng)引物及探針，金工并拓展了特異性、靈敏度、檢出限和定量分析等試驗。結果顯示：特異性試驗中，水短僅雞成(chéng)分檢測出現特異愛空性擴增，其他動物組織均未出現特異性擴增；梯度些文稀釋雞源性DNA模闆原液進(jì光刀n)行靈敏度評價，發(fā)現引物和探針靈敏度較高，可以檢測到100靜長 fg級的雞組分模闆DNA，且建立的标準曲線具有良了我好(hǎo)的線性關系，R2達0.是低99以上，定量準确；加工制品中的驗證試驗結果顯示，合成(chéng)的引物及拿志探針具有良好(hǎo)的适用性和定量檢測個雜能(néng)力。以上研究結果均說(shuō)明鐵通，此引物和探針适用于市場食品中雞源性成(chéng)分麗的的定性、定量檢測。本研究爲今後(hòu)TaqMan實時(sh錢科í)熒光PCR法不同動物源性引物和探針的黑很自主研發(fā)奠定了基礎。Quantitative Detecti低唱on of Chicken-derived Compone票在nts inFood Based o外草n Real-time PCRAt present，t短子he identification of 門討meat products is emphatically studied得多 in the field of food safety. In o答線rder to establish a quantita車但tive detection method for chicken-deriv理小ed components i音鄉n food，primers and probes 行動were synthesized according 書能to current standard of inspection and 時這quarantine（SN/T 2727—2010），and specific長秒ity test，sensitivi東朋ty test，detection lim問志it and quantitative analysis 和媽were expanded. The results showed 錢房that the specific amplifica我吃tion was found o科低nly in chicken-derived components in用要stead of other animal tissues；gra章件dient dilution of c很新hicken-derived DNA template were us報妹ed to evaluate the sensitivity，and多讀 it was found that the sensitivi人月ty of primers and probe wa花司s high，which could朋少 detect the DNA template at 100件喝 fg level，and a做雨 good linear relationship wa地喝s observed in the s月廠tandard curve，R2 could r生歌each above 0.99 with an accurate q雨妹uantitative detection. For the proce喝國ssed products，t自藍he synthesized primers and probes were 飛對with good applicability and quantita樂但tive detection capacity. It 紅件was concluded that the primers and雜日 probes were suitable for qualitative 醫很and quantitative 體雨detection of chicken-derived compo樂好nents in food markets. A foundation w樹制as laid for the indepe間科ndent development of different ani跳黃mal-derived primers and probes姐姐 of TaqMan real-ti著些me PCR in the future.計體全文下載鏈接：https://kns.cnki.net/KCMS/det開時ail/detail.aspx?dbcode來鐵=CJFQ&dbname=CJFDAUTO&fi店兵lename=ZGDW202008023&v=MDU5MDFYMUx個拍1eFlTN0RoMVQzcVRyV00xRnJDVVI3cW務區ZiK2RtRnlqa1dyN0JQeXJQZWJHNEhOSE1wND月呢lIWjRSOGU=
2020-08-20
豬血凝性腦脊髓炎病毒實時(shí)熒光RT-PCR檢測方法的建立與應用
爲建立一種(zhǒng)快速、準确、常規的豬關錢血凝性腦脊髓炎病毒（porcine hemagglutinating ence一錢phalomyelitis virus，PHEV）檢測方法，根據P公雜HEV N蛋白基因序列，設計１對(duì)引物和１條探針房可，建立了PHEV實時(shí)熒光RT-PCR檢測方法。該方法特異性好城兵(hǎo)，與豬繁殖與呼吸綜合征病毒（PRRSV來黑）、豬僞狂犬病病毒（PRV）、豬流行性腹瀉病毒（PEDV）、豬傳染錯費性胃腸炎病毒（TGEV）、豬瘟病毒（CSFV）和豬圓環病毒2型暗民（PCV2）均無交叉反應；用陽性質粒Puc不熱57-PHEVn評估其敏感性，發(fā)體兵現檢測下限達10-6 ng/μL（224拷貝/μL）。采集233年議份發(fā)病豬組織樣(yàng)品進(jìn)林懂行臨床檢測，發(fā)現該方法與巢式RT-PCR的符合率達98.3%，準确篩睡年查出巢式RT-PCR并測序陽性的樣(yàng)品14份，且檢出率高于巢式開遠RT-PCR。結果表明，本方法敏感、特異，可用于PHEV臨床樣(yàng)文不品的檢測。Establishment and Application 東體of a Real-time RT-PCR Assa能南y for Detection of Porcine Hemagg知靜lutinating Enceph舊公alomyelitis VirusIn order to establish花行 a rapid，accura妹呢te and routine me多市thod for detection of porci區上ne hemagglutinati拍通ng encephalomyelitis 道跳virus（PHEV），a pair of primers and光河 a probe were designed based熱有 on the N gene se公兒quence of PHEV，and a real-t不熱ime RT-PCR assay was develo土錯ped，which was wi作空th good specificity，an讀又d failed to crossly react with購區 porcine reproductive and resp視器iratory syndrome virus（PRRSV），porcine書子 pseudorabies virus（內煙PRV），porcine epidemic d身弟iarrhea virus（PE生討DV），transmissibl刀好e gastroenteritis virus（TGEV），c就弟lassical swine fever virus（CSFV）and po愛要rcine circovirus type 2（PC中頻V2）. The sensitivity of the met白知hod was evaluated b影暗ased on positive plasm場照id Puc57-PHEVn，and it was found相器 that the limit detection was 10-6 土男ng/μL（224 copies/μL）.業到 A total of 233 制靜swine tissues were tes下車ted by the method，and i廠一t was found that 14 positive姐視 samples were detected票算 out，and the coincidence rate慢那 with the nested 煙微RT-PCR was 98.3%，but the detection r黑區ate was higher than 如火that by nested RT-PCR.都市 It was concluded that t火東he method could be us請暗ed to detect PHEV clinical samples 時光benefited from it短雪s high sensitivity and specificity..還體全文下載鏈接：https://kns.cnki.net/KCMS少光/detail/detail.asp西和x?dbcode=CJFQ&dbname=CJFDAUTO&filename=也白ZGDW202008022&v=MzAwOTVUcldNMU學他ZyQ1VSN3FmYitkbUZ5am章飛tWNzdLUHlyUGViRzRITkh制有NcDQ5SFpvUjhlWDFMdXhZUzdEaDFUM3E=
2020-08-20
布魯氏菌S2疫苗陰道(dào)途徑免疫羊的安全性初步研究
布魯氏菌病（簡稱布病）是一種(zhǒng)由布魯氏菌引鐘子起(qǐ)的嚴重危害人和動物健康的人獸共患病工行。疫苗免疫是畜間布病流行率較高地區控制該病的重要手段。遼甯省建昌縣坐司經(jīng)過(guò)多年實踐子北，創新出一套免疫保護效果較好(hǎo妹子)的陰道(dào)途徑免疫S2疫苗的新型家畜免疫技術。爲了解該技術對(duì些科)家畜的安全性，本研究采用該技術免疫了3隻山羊，在免疫1個月少綠後(hòu)檢測其抗體滴度、體内含菌量，觀察脾髒和肝髒的病理變化。結果顯開文示，3隻免疫山羊均抗體陽轉，所有采集的髒器均未分離到布魯氏菌，且了跳脾髒和肝髒未出現布病相關病理變化。初步研究表明，通過(guò)陰道(dào)懂風途徑對(duì)羊進(jìn)行站冷S2疫苗免疫是安全的。Preliminary 現視Study on the Safety of Brucella V分拍accine S2 Strain on Goats Imm玩業unized via Vagina Brucellosis i美也s a kind of zoonosis ca月媽used by Brucella，which seriously e說動ndangers human and animal h雪哥ealth，and is mainly contr科日olled through vaccination in area就笑s where animal brucellosis is preval個要ent. After practices for se紙唱veral years，a new immune高算 technology（immunization via va新身gina）was developed 知影in Jianchang County of Li鄉雨aoning Province，a視拿nd good immune effects have been 要慢achieved. In order to make clear the s麗答afety of the technology on animals錯藍，3 goats were vaccinated，one month lat爸信er，their antibody tit火南er and bacterial content were detected 厭南and the pathologica城跳l changes of their s議業pleens and livers were obs睡影erved. The results showed that 子人the antibodies in the 金通three goats turned to be positiv議就e，no Brucella was isolated fro亮呢m all the organ快地s collected，and no pathological 紅笑changes related to b答事rucellosis were found in splee人這ns and livers. These results p答熱reliminarily indicat們子ed that it was safe to i腦我mmunize goat with Brucella vaccine用到 S2 strain via the 就術vagina.全文下載鏈接：https://kns.cnki.net/KC亮亮MS/detail/detail.aspx?dbcode=CJFQ&db身樂name=CJFDAUTO&f訊樹ilename=ZGDW202008019&v=M們日jU3NjBlWDFMdXhZUzdEaDFUM3FUcldNMUZyQ1V外熱SN3FmYitkbUZ5amtVN3ZJUHlyUGViRzRI志快TkhNcDQ5RWJZUjg=
2020-08-20
化學(xué)發(fā)光免疫分析技術在動物疫病檢測中的應用
化學(xué)發(fā)光免疫分析（chemiluminesecence男可 immunoassay，CLIA）是用于疫病檢測的一種(z下雜hǒng)免疫檢測技術，是基于免疫反應原舞男理，結合化學(xué)發(fā)光檢測技術而建立的。與放射免疫分析、酶免疫分析妹遠等标記免疫技術相比，具有無放射性污染、靈敏度高和特異性強的特點，已被(b她視èi)廣泛應用于疫病流行病學(xué)調查、診斷、藥物分析以及微生物工中檢測等方面(miàn)。本文簡要介紹了CLIA技術的來海基本原理及發(fā)展，從細菌、服愛病毒和寄生蟲檢測方面(miàn)，綜述了近年來該技術在動物疫朋鄉病檢測中的應用，提出該技術正向(xiàng)高特異性、高靈敏度、高通量制站、快速和自動化檢測的方向(xiàng)發(f短吃ā)展，這(zhè)爲今後(hòu)CLIA在動物疫病檢測中中章的應用研究提供參考。Application of Chemiluminesc舞煙ence Immunoassay in Detecti公村on of Animal Diseases As a kind of i去暗mmunological technolog雨筆y for detection 兵微of animal diseases，chemilumin的雨escence immunoassay（CLIA）is developed i歌機n combination with chemilumine愛購scence detection technology based on如相 the principle of immune 會機response，which is 我玩characterized by high讀嗎 sensitivity and good specificity wi兵兒th no radioactive pollution compare務少d to the immuno討是labelling technology 關站such as radioimmunoassay（RIA）and en花草zyme immunoassay（EIA），and has been 看我widely used for epidemio術我logical investigation an就什d diagnosis of animal diseases a請湖s well as drug analysi訊動s and microbial detection. In th個音is paper，the basic principle and 黃錯development of CLIA t厭路echnology were introdu快會ced，and its application i就就n animal disease d空白etection in recent years was summ說開arized from the到木 aspects of bacteria，v男數irus and parasite detectio用費n. It was shown that chemilumin黃一escence immunoassay was tur媽吧ning to be a rapid and automatic detec月自tion technology 麗你with high specificity，high sensi在黑tivity and high throughput. 草做This study would provide some ref了飛erence for application of CLIA in裡讀 animal disease dete家得ction in the future. 全文下載鏈接：ht錢線tps://kns.cnki.net/KCMS/detai放制l/detail.aspx?dbcode=CJFQ&dbname=近但CJFDAUTO&filename=ZGDW2020080船遠18&v=MDMwNDkrZG1GeW5oV3JyT1B5cl謝妹BlYkc0SE5ITXA0OUViSVI4ZVgxTHV4WVM3RGgxV計還DNxVHJXTTFGckNVUjd睡放xZmI=
2020-08-20
水生動物冠狀病毒研究進(jìn)展
冠狀病毒在自然界中普遍存在，可感染多種(zhǒ化美ng)哺乳動物和鳥類，給畜牧業生産安全和公共衛生安全帶來嚴重威脅。自北京新發(從音fā)地批發(fā)市場從切割進(jìn)口外店三文魚案闆上檢測到人新型冠狀病毒（水街SARS-CoV-2）核酸以來，三文魚以及水生動物冠狀病毒與SARS-年喝CoV-2的關系成(chéng)爲公衆關注的焦點。草器本文詳細描述了白鲸、寬吻海豚、三文魚等水生動物感染冠狀病毒後(hòu)白鐘的臨床症狀，以及病毒基因組結構特征，并對(duì)水生動物冠狀病毒與SAR員森S-CoV-2進(jìn)行了遺傳演化分析，公得結果顯示目前已知的水生動物冠狀病毒長事屬于γ冠狀病毒屬和Alphale動家tovirus屬，而SARS-CoV-2屬于β冠狀病毒屬，二者基我友因組和主要基因（1ab、S、E、M、N）核苷酸同源性僅和紅爲43.7%~54.1%，可初步排除SARS-CoV-2來源于已知水玩匠生動物冠狀病毒的可能(néng)性。本文路東通過(guò)系統闡述水生動物冠狀病毒感染狀況，旨在進(jìn)一步提高雪習公衆對(duì)水生動物冠狀病黑師毒的認知，爲科學(xué)防控冠狀病毒提供理論依車關據。 Research Progress on Cor區科onaviruses in Aqua化要tic AnimalsCoronavir相相us is widely prevalent in n中內ature and may infect a variety of mamma銀線ls and birds，which poses a ser村現ious threat to the safety of ani知化mal husbandry and public計綠 health. The relationship be熱空tween coronaviruses in aquatic 雜數animals including salmon and severe acu技大te respiratory syndrome coronavirus 2謝兒（SARS-CoV-2）has been concer什鐘ned by the public since the n嗎長ucleic acid of SARS-CoV-2 花秒was detected on the cutt到志ing board for i生費mported salmon in 家道Beijing Xinfadi 議河wholesale market樂體. In this paper，the clinical裡音 symptoms of infected aquatic animal門視s such as belugas，bottlenose 長城dolphins and salmon，as w要媽ell as the structur醫民al features of viral genome wer錯兵e described in 訊們detail，the genetic evolution 金畫of coronaviruses in aquatic 短坐animals and SAR東著S-CoV-2 were anal去歌yzed. The results showed that the志兵 known coronaviruses in aquatic animal冷校s belonged to Gamma飛兒coronavirus and A就習lphaletovirus，while S不媽ARS-CoV-2 fell into Beta日線coronavirus，and the nucleotide hom志錢ology of their genomes and major g兵窗enes（1ab，S，E，M and N）was 視計43.7% to 54.1%，which co路鄉uld exclude any possibility of SARS-C跳很oV-2 originating from the known c拍員oronaviruses. The infection 話樹status of coronaviruses in aquatic電是 animals was systematically desc年子ribed in this paper，hoping 問的to further improve public 舊拍awareness and provide theoretical ba說身sis for scientific prev草站ention and cont美物rol of coronavirus. 全文下載鏈接：放微https://kns.cnki.net/KCMS/deta習術il/detail.aspx?dbcode=CJFQ&秒民dbname=CJFDAUTO&filename=Z對話GDW202008017&v=MzI2NzkxRnJDVVI3cWZiK愛行2RtRnluaFZML0tQeX月明JQZWJHNEhOSE1wNDlFWTRSOGVYMUx1eFlTN0R船又oMVQzcVRyV00=
2020-08-20
雲南省藍舌病病毒“曆史毒株”的遺傳特征
20世紀90年代，本實驗室從雲南省哨兵動物采集的血液中分離到7種(志北zhǒng)血清型藍舌病病毒（bluetong山那ue virus，BTV）。但明信這(zhè)些毒株的遺傳特征至今仍為紙不清楚，因而阻礙了我國(guó)的章務BTV演化曆史分析。爲掌握雲南省早期流行BTV毒株的遺傳特征，吧河對(duì)1995—1997年雲南省分離的25子服株BTV毒株的基因組節段2、3、7和10（Seg-2、-3、-7、不說-10）進(jìn)行一步法RT-PCR土低擴增、測序以及序列分析。Seg-2序列分析顯示，1995—議藍1997年雲南省分離到的7種(zhǒ話車ng)不同血清型（BTV-1、-2、-3、-4店謝、-12、-15、-16）BTV毒株，分屬A、B要影、G、H、I和J等6種(zhǒ區水ng)基因型；BTV-12型毒株的Seg-2爲西方地域型公河，其餘毒株的Seg-2則屬于東方地域做自型；Seg-3和Seg-10序列分析顯示，西唱25株BTV毒株均爲東方地域型；Seg-7序列分頻懂析顯示，1株BTV-2型、2株BT作在V-12型和1株BTV-16型毒株屬于西方地域亞型，并與南非和業銀荷蘭BTV毒株的Seg-7具有最近的親緣關系，其餘毒株則均爲農身東方地域型。上述結果表明，雲南省存在多種(zhǒng)血清筆林型BTV的流行，而Seg-2和Se河鄉g-7基因重配毒株的發(fā)現司對，提示國(guó)外的西方地域型毒株已侵入雲南省，并在傳播過視藍(guò)程中與我國(guó)毒株發(fā)生了基因下些重配。本研究爲我國(guó)BTV的演化分析與溯源研究提供在外了數據參考。 Genetic Characteristics of“Hi體刀storical Strains”of Bluetongue 關玩Virus Isolated fr物懂om Yunnan Provinc化坐e In the 1990s，7 serotypes of bluetongu森兵e virus（BTV）were isolated from blood s場風amples collected fr資玩om sentinel animals in Yunnan Provi現錯nce by our labora好月tory. However，the gen近歌etic characteristics of these strains章長 were still not clear，which hindered國訊 the study on the evolutio窗相nary history of BTV in子明 China. In order to mak車來e clear the genetic characteristics看廠 of BTV strains spread early i門外n Yunnan，one-step RT-PCR森知 amplification，著在sequencing and sequence analysis were c嗎子arried out for gene兒了 segments 2，3，7 下靜and 10（Seg-2，-3，-7 and-10）of 25 BTV st要靜rains isolated in the province from能子 1995 to 1997. It was found t到作hat，by Seg-2 sequence analysis，Sev拍外en different BTV serot聽音ype strains（BTV-1，劇銀-2，-3，-4，-12，-15 and -16）we木見re isolated，whi讀話ch fell into 6 nu那商cleotypes of A，B，G，又笑H，I and J，respectively；the Seg-2 of B鐵湖TV-12 fell into Western topotype，but ot得內her strains belonged t房知o Eastern topotype；it歌木 was showed tha我來t，by Seg-3 and Seg-10 sequence anal間文ysis，25 strains of BTV belonged to Ea著校stern topotype；and by Seg-林湖7 sequence analysis，one strain of BTV-2從他，two strains of BTV-12 and one strain 紅報of BTV-16 were clustered into 人高Western topotype，and shared the clo讀雪sest genetic relationship wi視山th Seg-7 isolated from South Af飛風rica and Netherl為花ands，and the other 報藍strains belonged 土房to Eastern topotyp體紙e. It was concluded that multiple serot女河ypes of BTV wer爸兵e prevalent in Yunn身雨an Province，and 看什foreign Western topotype strains下如 had been introduced into問說 the province and reassorted with Ch務舞inese strains in the pro商跳cess of transmi分車ssion as Seg-2 and Seg-7 gene re多拍assortative isolat拿影es were detected. Some data was p兒上rovided for evolution ana路時lysis and traceability st南頻udy on BTV in China.全文下載鏈接：https愛靜://kns.cnki.net/KCMS/de農國tail/detail.aspx?db件子code=CJFQ&dbname=CJFDAUTO&f外這ilename=ZGDW202008007&v=MTYzNDg3RGg來照xVDNxVHJXTTFGckNVUjdxZmIrZ愛謝G1GeW5oVnI3TlB5農路clBlYkc0SE5ITXA0OUZZNFI4ZVgxTHV村音4WVM=
2020-08-20
馬泰勒蟲RAP-1基因的原核表達及蛋白性質分析
棒狀體相關蛋白1（rhoptr作話y-associated protein 1，RAP-1）是由棒狀體分泌的與錢海侵襲宿主細胞相關的蛋白。本研究以頂複門原蟲RAP-1基因爲靶标，用最大計山似然法構建了巴貝斯屬、瘧原蟲屬、泰勒蟲屬的RAP-1基因進(j匠是ìn)化樹；應用軟件預測分析RAP-1們雪蛋白的理化性質、親疏水性、跨膜區以及二、三級結構，姐西并表達純化了馬泰勒蟲（Theil城習eria equi）RAP-1重組蛋白。結果顯報林示：巴貝斯屬不同蟲株相聚類，測定序列與日鐘馬泰勒蟲聚爲一支，又與惡性瘧原蟲相聚類，表明親緣關系較近。馬泰勒蟲RAP-這雜1蛋白理論等電點爲9.85，半衰期爲30 h，總平均親月拿水性（GRAVY）爲-0.368，無跨膜區；RA新姐P-1蛋白二級結構中，α-螺旋、β-折疊、β綠短-轉角和無規卷曲占比分别爲38.5%、15.坐村5%、27.8%、18.1%；構建的三級結構立體展現了RAP-高樹1蛋白形态。成(chéng)功構建了原核表達載體pET-32a-RAP短城-1；該重組蛋白主要以包涵體形式存在，蛋白大小爲下微65 kDa。RAP-1蛋白可通過(guò)原核表達系統高效表達，純化後(小子hòu)的産物能(néng)被(bèi機鐘)馬泰勒蟲标準陽性血清識别，具有良好公資(hǎo)的反應原性。本研究爲深入了解馬泰勒蟲入侵宿主的機制機理錯開以及RAP-1蛋白的功能(néng)研究奠定醫離了基礎。Prokaryotic Expression of RAP-1 飛樹Gene of Theileriaequi andCh友有aracteristic Analysis on海吃 the Encoded ProteinRh街謝optry-associated pro是麗tein 1（RAP-1）is a明短 protein related to invading the ho看得st cells and secreted by t校男he rhoptry. Targeting at the R學購AP-1 gene of Protozoan protozoa，the了員 gene evolution trees 事件of Babesia，Plasmodium and Theiler白資ia were constructed by the max弟友imum likelihood me民朋thod. The physica店中l and chemical properties，hydrophobi吃錢city，transmembrane zone，se劇日condary and tertiary structures of那秒 RAP-1 protein were predicted by什看 software，and the recombinant RAP-1 道城protein of Theileria eq裡雪ui was expressed and purified. The 子討results showed that the gen農時us of Babesia was clustered 男舊together，its sequence and that of匠友 Theileria equi belonged 商數to the same branch and clu公務stered with the genus o事草f Plasmodium falciparum，indicati木東ng that their genu章空s was closely related. Th還白e theoretical isoel房城ectric point of RAP-1 我都protein was 9.85，the half-life wa水內s 30 h，the total村業 average hydrophilicity was -0.368照對，and there was no transme問吧mbrane zone. The secon上用dary structure α-helices，β-sheet暗下s，β-turns and random厭秒 coils of RAP-1 protein ac生上counted for 38.5%，15.5%，27.8% and一黑 18.1%，respectively. The form o輛短f RAP-1 protein was煙業 shown by the three-level s很朋tructure，and the prokaryotic expre外器ssion vector pET-32a-RAP-1 was succ錯票essfully constr好飛ucted，the recombinant protein 影生mainly existed in the form of inclus你多ion body with the size of 65 kDa在雨. In conclusion，the RAP鐵草-1 protein could 司工be highly expressed by pr影木okaryotic expression system，and the pur西雨ified products could be identified b風街y the standard positive serum o車湖f Theileriaequi due to its good re上務actogenicity. Th去看erefore，a foundation was prov音子ided for further studyi歌遠ng the mechanis女在m of Theileriaeq要頻ui invading into hosts and 間廠the functions of RAP-1 protein.日服全文下載鏈接：https://kns.cnki.net/KCMS/detai資們l/detail.aspx?dbcode=CJFQ&dbname=個算CJFDAUTO&filename=ZGDW202007022&小妹v=MTU3NDdTN0RoMVQ吃機zcVRyV00xRnJDVVI3cWZZT2RvRnl6bFU3dlB為弟QeXJQZWJHNEhOSE1xSTlIWm們笑9SOGVYMUx1eFk=
2020-07-17
韓長(cháng)賦會(huì)見俄羅斯聯邦農業有動部部長(cháng)帕特魯舍夫
習近平主席特使韓長(cháng)賦出席委内瑞拉總統就(jiù)樂麗職儀式
韓長(cháng)賦在農業農村部非洲豬瘟疫情應急信得指揮部會(huì)議上強調：強化責任擔當 形成(chéng要厭)工作合力 共同打赢非洲豬瘟防控殲滅戰持久戰
豬流行性腹瀉病毒RT-LAMP可視化檢測方法的內美建立
動物衛生信息快報2019第17期
農業農村部副部長(cháng)于康震赴黑龍江督導非洲豬瘟防控工那子作時(shí)強調：壓實責任 落實措施 嚴是風防疫情擴散蔓延
北京市延慶區農業局（北京市延慶區動物腦務衛生監督管理局）2019年度公開(kāi)招聘體檢考察公告
中心組織開(kāi)展“過(gu如河ò)政治生日，憶入黨初心”主題黨日活動
2019農曆豬年 讓我們爲豬年慶祝并保護生劇看豬！
2019年第一期國(guó)家影愛官方獸醫師資培訓班在青島舉辦
