豬德爾塔冠狀病毒納米PCR檢測方法的建立
根據GenBank登錄的豬德爾塔冠狀病毒（PDC微山oV）毒株M基因設計特異性引物，建立了PDCo嗎都V納米PCR檢測方法并進(jìn)行了臨床樣(yàng)品檢測。學是結果顯示：該方法對(duì)标準品檢測的最低濃度可達102 cop在唱ies/μL，與其他幾種(zhǒng)常見豬病病毒站花無交叉反應；用該方法檢測國(g關電uó)内豬場62份腹瀉糞便樣(yàng)品，相比常規PCR方來城法，該納米PCR方法的檢出率更高，很商而兩(liǎng)種(zhǒng)方法對(duì)268份進(jìn錢鄉)境活豬糞拭子樣(yàng)品檢測均爲陰性。結果表秒學明，本研究所建立的納米PCR檢測方法适用于國(guó)内豬場的PDCoV海姐檢測和進(jìn)境活豬的監控檢測，器務可作爲PDCoV實驗室檢測的有力技術手段。Esta物慢blishment of Nanoparticle-as做個sisted PCR Assay for Detectio費東n ofPorcine Deltacor腦離onavirusIn this paper，a pair ofp來冷rimers were designed according to the 動電M gene sequence of porc外嗎inedeltacoronav慢有irus（PDCoV）that waspublished in跳姐 Genbank，then a kin得睡d ofnanoparticle-assis鐵舊ted PCR assay was establish了嗎ed. The results showed 章來that thestandard s男村amples with minimum concentr理場ation of 102 copies/μL could b話錯e detected by this method，and大但 therewas no any cross-reaction wi高雪th other common swine disease vi熱會ruses. Compared toregular刀化 PCR assay，the nan去從oparticle-assisted PCR assay船校presented a higher detection rate when下自 62 diarrhea fec看工es collected fromdome信工stic swine farms were tested. 能兵When 268 fecal swa自了b samples of imported livepigs we樂白re tested by both the two明男 methods，the detectionresu個行lts all showed negative. Therefore，刀學thenanoparticle-assisted PCR assay es資站tablished in the paper could be used in暗舊detecting PDCoV in domestic swine計南 farms and monitoring imported live p綠聽igs，which would be門看 a strong technical tool在車 for laboratory detection ofPDCoV土得. 全文下載鏈接：http://kn商兒s.cnki.net/KCMS/detail/37.1246.S.20190影請802.1405.040.html
2019-08-13
豬僞狂犬病病毒與副豬嗜血杆菌混合感染的綜合診斷及感讀相染毒株gE基因遺傳進(jìn)化分析
2019年4月，山東省新泰市某育肥豬場從某種(zhǒng)豬身厭場購進(jìn)了100頭5周齡仔豬。購進(jìn)1周後(hò開呢u)，有40多頭仔豬表現昏睡、嘔吐、拉稀等症狀如水并出現急性死亡，共死亡28頭，病死短紙率高達68%。對(duì)病豬進(jìn)行病理剖檢、組織病理學體吧(xué)檢查、病原學(xué)檢測等實驗室綜合診斷，最終确診爲僞狂犬病月畫病毒和副豬嗜血杆菌的混合感染。對(duì有開)分離的僞狂犬病病毒gE基因進(jìn)行測國就序和遺傳進(jìn)化分析，确定該分離株與著低近期流行株同源性較高，這(zhè)爲該病知現疫苗的選擇和高效使用提供了依據。這術基于診斷結果，制定了相應的防控措施，使疫情得到了迅速控制。民業本文還(hái)分析讨論了該病例的發(fā)生原因、技拍發(fā)病特點及病變特征，對(duì)類似病例的診日低斷防控具有一定的指導意義。Comprehensive D鐘林iagnosis of Mixed Infection with P舞放orcinePseudorabies Virus a短少nd Haemophilus parasuis and Phylo可又genetic Analysis 水看on the gEGene of Infected S年裡trainIn April 2019，100 piglets影視of 5-week old wer紙工e purchased from a breeding farm醫個 by a fattening farm in XintaiCity o森如f Shandong Province. 光黃Subsequently，some symptomssuch as紅很 lethargy，vomiti農費ng，diarrheaor ev購線en death appeared in more than 40 p睡朋iglets after one week能新，a total of 28 piglets died，場下with a mortalityrate of 68%，which wa電鐵s finally confirmed as the m拍白ixedinfection with pseudorabies vi雜有rus and Haemoph藍頻ilus parasuis through laboratorycompreh內動ensive diagnosis includin書快g pathological autopsy，histopathologi票做calexamination and etiological tes媽是t. Then the gE gen土湖e of isolated pseudorab年件iesvirus was sequenced and ana睡拍lyzed，and high homo什刀logy ofthe isolate with c西土urrent epidemic strain was found，他身whichprovided a basis for select子行ion and efficient application of t懂動he vaccine. Basedon the above短音 results，corresponding contro就民l measureswere formu影睡lated，and the outbreak was rapidlycont身行rolled. Furthermore，t器科he reasons，clinical charac錢土ters and pathological changes of the ca高雨ses werediscussed and analyzed in the 爸影paper，which were ofsign也醫ificance for diagnosis and con這門trol of similar cas生去es. 全文下載鏈接：http://kns.cnki.net/KCMS/和看detail/37.1246.S.20190802.14資火05.038.html
2019-08-13
4例禽戊型肝炎病例的實驗室診斷及ORF2基離你因遺傳進(jìn)化分析
本研究通過(guò)分子流行病學(南弟xué)診斷技術，分别從山東、遼甯、吉林、陝西4個地區表現肝炎症狀的微樂病雞中，采集肝髒和脾髒樣(yàng)品，利用RT-PCR方法，檢測禽戊型肝個照炎病毒（aHEV），并對(duì)aHEV-ORF2 基因片段進(近動jìn)行測序，利用MegAl我男ign進(jìn)行同源性比較。結果顯示：4個雞老照場的病雞樣(yàng)品均爲aHEV 陽性，aHEV-ORF2基因序列購作間同源性爲84.5%~94.5%，與已報道(dào)的4種(zhǒ商會ng)基因型同源性均小于84.0%，其中與基因1型的澳大利亞株和韓國(guó)刀日株同源性爲81.6%~82.8%，與基因2型的美國(guó紙兒)原型株和美國(guó)無毒株同源性爲80.9%~82.8%，與基因3什門型的歐洲株和中國(guó)株同源性爲80.西街8%~82.5%，與基因4型的匈牙利株和中店哥國(guó)台灣株同源性爲 81.4%~82.6%。ORF2基因進(jìn)外了化樹分析顯示：4株aHEV均屬于戊型肝炎B種(影雨zhǒng)，形成(chéng)獨立的基因分支，但不屬于已報道(dà慢工o)的4個基因型，而在一個新型獨立分支上理什，表明國(guó)内存在一種(zhǒn公會g)新基因型的aHEV流行。本研究爲我國(guó)aHEV的進(jìn是員)一步研究提供了基礎和依據。Labor光師atory Diagnosis of 說河four Cases of Avian 錯玩Hepatitis E andPhylogen和子etic Analysis on Its ORF2 GeneIn th友很e study，throughdiagno弟木stic technology of molecu森土lar epidemiology，liverand spl習時een samples were collected fro身呢m sick chickens with sympt靜來oms of hepatitisin four farms in Sh請的andong，Liaoning，Jilin and Sha遠雨anxi provinces，then avian喝廠 hepatitisE virus（aHEV） was detected銀小 byRT-PCR and the fragments of aHEV票高-ORF2 gene were sequenced. At last生農，the homology was compared by m船但eans of MegAlign. T熱場he resultsshowed that all samples f離媽rom the above f亮城our farms were positi歌外ve for aHEV，and the homology be明司tween the gene seq玩子uences ranged from 84.5% to94.短議5%. In addition，the homologi離土es of the four i關個solatedstrains with four alre店坐ady published g風化enotypes were all less than 84.0劇作%，specifically，the homologies with gen喝秒otypeHEV 1（Australian and Ko區謝rean strains），H樂樹EV 2（American prototype and Americanvi訊城rus-free strains），HEV 3（Eu開又ropeanand Chinese strains子遠），HEV 4（Hungarianand Taiwanese-chi金對nese strains）were 81.6% to 82.8喝分%，80.9% to 82.8%，80.8% to 82.5% a器會nd 81.4% to82.6%，respectively. Accordi舞白ng to the analysis of evolut筆低ionarytree of ORF2 gen鄉妹e，the four strains of aHEV 站去allbelonged to B type of hepatitis E，i美爸nstead of the fourreported genotypes，w要空hich formed a new separate genebranc遠務h，indicating a new genotype was preva離歌lent inChina. The research wou從報ld provide some ref文做erences and foundation for furth服吧erstudy of aHEV in China. 全月是文下載鏈接：http://kns.cnki.n師些et/KCMS/detail/3信去7.1246.S.20190802.1405.036.ht船唱ml
2019-08-13
重組禽流感病毒二價滅活疫苗（H5N都黑1 Re-8株和H7N9 H7-Re1株）對(duì)雉免小答疫效果的研究
爲評估重組禽流感病毒二價滅活疫苗（H5N1 Re-8資匠株和H7N9 H7-Re1株）對(du吃知ì)雉的免疫保護效果，選取30日齡的雉，按照免疫程序首免後(hòu)21 議場d，加強免疫1次，分别于接種(zhǒng)前和接種(zh風外ǒng)後(hòu)定期采血，分離血清，檢測H5、年朋H7亞型HI抗體滴度。結果顯示，免疫前，雉血清中H5、H7亞型老個HI抗體滴度均爲0；免疫後(hòu) 14 d，H個錯5和H7亞型抗體平均滴度分别達到7.0log2和7.07log2，49水金 d後(hòu)平均滴度達到峰值，分别爲8.04log2和9.20log場遠2；H5亞型免疫有效期可達105 d以上，H7亞型可達189 d以上。結水妹果表明，重組禽流感病毒二價滅活疫苗（H5N1 Re明道-8株+H7N9 H7-Re1株）對(duì)雉具有良好(hǎ資照o)的免疫保護效果且安全性良好(hǎo)，但疫苗免疫49 d後(厭錢hòu)，H5、H7抗體效價逐漸下降雪知，133 d後(hòu)明顯下降，群體免疫合格率低于著議70%，因此生産中推薦，30日齡首免後(hòu)，51日齡和5我生月齡各加強免疫1次。本研究爲珍稀禽的禽流感免疫疫苗選擇和程序制相業定提供了參考。Study on the Immune Effects 工音of Recombinant Avian Infl靜會uenza VirusBiva市白lent Inactivated Vaccines（H5N1 Re-8+H7中服N9 H7-Re1 Strain）in Pheas影討ants In order to evaluat歌器e theimmune effects of r廠頻ecombinant avian influe通看nza virus（A/V） bivalentinactivated v土們accines（H5N1 Re-8+H畫少7N9 H7-Re1 strain）in phe間高asants，30-day-old pheasant新議s wereselected for fi志吃rst vaccination，followed by booster跳女vaccination after 21 days according to聽問 immune procedures，and their blo年木od samples were collected r理鐵egularly before and after那笑vaccination respectiv舞都ely to test the titers of HI an制妹tibodies against H5 and H7sub科冷type AIV. The re資聽sults showed that，before vaccin但雨ation，the titers of HI anti科他bodies against H5 and H機低7 subtypes in the ser歌但umof pheasants were 0；and the averag計醫e titers of H5 and H文舊7subtypes could r城信each to 7.0log2 and 7.07log2 r信車espectively after 14 d麗做ays，then increased to the問家 peak after 49 days，whichwere 8.04lo地為g2 and 9.20log2 respectively；the immune這亮validity for H5 subtype A自場IV could remain for 105 days or above，a雜分nd that for H7 subtype子拍 could be up to 189 days，which indicat分視ed the immune effects an事市d security of bivalentinactivated vacc廠月ines were good. H志很owever，the titers 鄉著ofH5 and H7 antibodies船器 decreased graduall少但y after 49 days of vaccination，但劇then decreased significantly 飛舞after 133 days，andthe qualified 影懂rates at herd level was 鐵煙less than 70%. Therefore，it書國 was suggested that，after the f子到irstimmunization at th船船e age of 30 days，th店自e reinforcedvaccination should be c問分arried out at the ages of 51 days and間要 5 monthsrespectively. I銀就n conclusion，the research wo討但uld providesome references for se放媽lecting vaccines（文對for rare andprecious河海 birds）and developing relevan件店t procedures.全文下載鏈接：ht街錯tp://kns.cnki.net/KCMS/detai地女l/37.1246.S.20190802.1405.034.html
2019-08-13
廣西動物防疫物資信息管理系統的構可和建與應用
爲實現廣西動物防疫物資的信息化管理，适時(shí)哥朋掌握廣西動物防疫物資的流轉狀況，廣西壯族自治區動物疫病預防控制中愛雪心開(kāi)發(fā)了“廣西壯族自治區動物防疫物資信車紙息管理系統”。該系統爲B/S架構，采用JSP技術和MySQL數據庫平台。船校該系統建立後(hòu)，廣西各級動物疫病預防控校間制機構均可通過(guò)互聯網登錄車議，及時(shí)存儲和統計動物防疫物資信息為了數據，全程監控和實時(shí)了解物資流轉情況。該系統爲廣西動物防疫物資的規快子範化管理提供了重要的技術支持，也爲全區重多務大動物疫病防控提供了堅強的後(hòu)離兒勤保障。Establishment and Application of有雜 Information Management System of Anima計年l Disease Control Materi妹河als in GuangxiIn order to rea知紅lize information-based management o頻很f materialsfor animal disease pr謝機evention and control in Guangxi，an的妹d to recognize the movement status of視呢 thematerials in time，an“informationman南些agement system of animal 能學disease control materials”withB/S fram書時ework was developed 爸房by Guangxi Zhuang什民 Autonomous Region C黃去enter forAnimal Disease 家線Prevention and Control by means of J音雪SP technology and MySQ秒近Ldatabase platform. With the森會 system，data related t場還oanimal disease control materi做校als could be stored a不著nd summarized on line byrelev匠畫ant authorities at all 來拍levels in Guangxi，so thatall movement快費 status of the mate理信rials could be mo門家nitored in real time.用時 It wasconclude一草d that important technical s身車upports were provided for stand錯志ardizedmanagemen秒生t of animal disease co吃和ntrol materials in Guang房自xi by the system，and the st明門rong logistical supports were al上答so guaranteed for thecontrol of maj生多or animal diseases. 全文下載鏈接：校請http://kns.cnki.樹得net/KCMS/detail/37.1246.S.20190802.問兒1405.024.html
2019-08-13
恩諾沙星及其代謝物在方正銀鲫組織中的代謝及消光機除規律
近年來水産養殖中的常用藥物恩諾沙星在鲫中的殘留問題短遠比較突出，但目前國(guó)家未對(duì)其在鲫電舊中的休藥期作出明确規定。爲做到鲫養殖中科學(xué和黃)合理使用恩諾沙星，在12~15 ℃水溫條件下，以60 mg/國空kg.b.w.劑量，對(duì)體質量爲（250±10）g的健康方正銀鲫他海（Carassius aurat計坐us gibelio）灌服恩諾沙星要花；在灌服後(hòu)0~120 d内不間斷采樣(yàng)，爸新用高效液相色譜-串聯質譜儀檢測，研究恩諾沙星及其代謝産內技物環丙沙星在方正銀鲫鰓、血漿、肌肉、皮膚、肝髒和腎髒中的代謝及消除規律，從外購而爲恩諾沙星在方正銀鲫中的休藥期制定提供參考。結果顯示：灌藥後(h和飛òu)，恩諾沙星在方正銀鲫血漿、肌肉、肝髒、鰓、腎髒和皮膚中的達峰時(他路shí)間tmax分别爲9、18、24、24、48和48 h，腎髒、肝山明髒、肌肉、血漿、鰓和皮膚中的達峰濃度c在農max分别爲30.490、21.372、18.715、16.636熱裡、15.157和11.663mg/kg；皮膚暗唱中的消除半衰期t1/2β最大，爲知時338.2 h，血漿中最小，爲93.303姐媽 h；代謝産物環丙沙星的代謝及消除趨勢與恩諾沙星大緻相同，血漿中達峰值産生最早哥都，tmax爲18 h，腎髒中的峰濃度最大，cmax爲547.26 μg/k廠商g。結果表明，方正銀鲫以單次口灌60 mg/kg.b.w.劑量恩諾沙星，皮膚美能和肌肉中的恩諾沙星和環丙沙星總量需要1200度日才能(néng)滿足限量要求窗子。本研究爲恩諾沙星在鲫魚養殖中的科學(xué)使用提供了數據支撐。E坐做stablishmentof a Rea習鐵l-time RPA Method for Rapid錯站 Detection of Lymphoc道離ytic ChoriomeningitisVirusInre相呢cent years，the residues of Enrofl對習oxacin that has been comm年月only used inaquacultu店秒re have become more serious in C錯票arassius auratus 舞路gibelio，however，no any clear regulation高南 on itswithdrawal period has南吧 been developed by the state. In order弟兒 to scientifical公和lyuse the Enrofloxacin in 化但carp breeding，the fol討好lowingexperiment was carried out. Hea現火lthy Carassius auratu們你s gibelio with body mass of（250±10）g w們也as fedwith Enrofloxacin at 木他a dose of 60 mg/kg.b.w. under the wa少間ter temperature of 12to 15 ℃外雨，and then incessa著花ntly sampled within 0 to 12哥兵0 dto test by high perfo樹計rmance liquid chromatog分要raphy-tandem mass spectrometry器科（HPLC-MS），hence the metabolism m相子echanism wasstudied，as well as the e喝土limination rule科筆 of Enrofloxacinand its metabolite（C女生iprofloxacin）in gill，plasma西商，muscle，skin，liv唱頻er and kidney，which could provide s行我ome references 去請for defining its withdrawalperiod都謝. The results showed that，after ad樂畫ministration，the peak time（tmax）ofEnrof低長loxacin in plasma，muscle，liver，g厭新ill，kidney and skin was 9，18，24匠美，24，48 and 48 h，respectively and th站器e peakconcentration（cmax）was 16.6金術36，18.715，21.372，15.157，30.49的友0 and 11.663 mg/kg，res通東pectively. Theelimination half-l分微ife（t1/2β）inskin was the匠歌 maximum，which was 338.2 h，an醫到d in plasma it wa舞資s the minimum（93.303 都離h）. The metabolism and兒站 elimination trend o熱鐵f Ciprofloxacin was similarto that of吧又 Enrofloxacin，specially，the peak va鐵關lue first appeare議筆d in plasma with南算 tmax of 18 h，and the peak concentra聽章tion reached the maximum in kidney 睡那with cmaxof 547.26 μg/kg知妹. Therefore，aconclusion was gi很對ven that it would take mo腦風re than 1 200 degree-days for thet跳知otal amount of 雜店Enrofloxacin and Ciprofloxaci分房n in skin and muscl林生e to reach ther秒坐equired limit if Carassius auratus gib子海elio was orally administered wit熱麗hEnrofloxacin at a dose of 木明60 mg/kg.b.w each time. There放時fore，some data supports were prov外文ided by this study fo日分r scientific use ofEnrofloxacin in carp水短 breeding. 全文下載鏈接：http://kns.cnki志如.net/KCMS/detail/37.1246.S土呢.20190703.1252.042.ht靜關ml
2019-07-31
兔源肺炎克雷伯氏菌重組酶聚合酶擴增檢測方法的建立
爲建立一種(zhǒng)檢測兔源肺炎克雷伯氏菌（Kleb化化siella pneumoniae機章）重組酶聚合酶擴增方法（recombin問歌asepolymerase amplification，農微RPA），根據肺炎克雷伯氏菌的phoE基因保守序列設計飛慢引物，擴增片段大小爲277 bp，并對(duì)反應條件進到要(jìn)一步優化，最終建立了适宜于快速準确檢測兔源肺炎克知些雷伯氏菌的重組酶聚合酶等溫擴增方法。該坐報方法可特異性檢測出兔源肺炎克雷伯氏菌，最低檢出外業細菌量爲8.3×101 CFU/m為東L，靈敏度比傳統PCR高100倍。本研究建立的肺炎克雷伯氏菌RPA女票檢測方法特異性強、操作簡單，從而爲生産中的兔源肺炎克雷伯氏菌現場快速檢務輛測提供了一種(zhǒng)新方法。Establishme新花ntof a RPA Assay for Detecting Kl很站ebsiella pneumon什了iae of Rabbit Orig唱農in In order to establish a recombinase計喝polymerase amplification（RPA）a亮水ssay for detecting Kleb事畫siella pneumonia of rabbit origin河去，a pair of primers were designed based要爸 on the conservative sequenceof PhoE g費做ene of Klebsiella pneumoniae，and th間離e fragmentwith the length of 277 bp was子花 amplified，then thereacti兒睡on conditions wer用劇e further optimized，fina相水lly，the RPA assay was establishe也離d，by which，the Kl影道ebsiella pneumoni呢的ae of rabbit origin could be specifi費朋callydetected with the分離 minimum bacteria音亮 of 8.3×101 CFU/mL，and its se說土nsitivity was 100 times higher than t不件hat of traditio謝黑nalPCR. Therefo務我re，the method es書美tablished in the 我坐study hadcharacteristics 費火of strong specifi畫校city and simple operation，which 議相would provide a new m一子ethod for rapid detection of Kleb鐘爸siellapneumoniae費得 of rabbit origi服新n. 全文下載鏈接：http://kns.cnki.net/KCM月船S/detail/37.1246.S用舞.20190703.1252.036.html
2019-07-31
淋巴細胞脈絡叢腦膜炎病毒實時(shí)熒光RPA快速錯相檢測方法的建立
針對(duì)淋巴細胞脈絡叢腦膜炎病毒（lymphocyt間司ic choriomeningitis virus，LCMV）基因保守序能可列，設計特異性引物和熒光探針，建謝對立了LCMV實時(shí)重組聚合酶等溫擴增（rea國知l-time recombinasepolymerase amp為鄉lification，real-time 作美RPA）檢測方法。該檢測方法具有良好(hǎo)的特異性，與LC志拍MV同步檢測的其他鼠病毒均未出現交叉反應；該檢測方法具有與RT-PCR拿紅一樣(yàng)的高敏感性；通過(guò)對(duì)LCMV感染鼠廠老組織樣(yàng)本和LCMV陰性鼠血清樣(yàng)本的檢測，證實建立的實這市時(shí)熒光RPA方法具有良好(hǎo)的穩定性和可靠性。與現事船有的核酸檢測方法相比，該實時(shí)熒光RPA檢測行市方法在核酸上樣(yàng)20 min内即可判讀結果，可滿足齧校書齒類動物LCMV快速檢疫需要。Establ北科ishment of a Real-time 女睡RPA Method for Rapid Detection ofLymp朋動hocytic Choriom上國eningitis VirusIn this paper，specificp玩雪rimers and fluore海行scent probes were designed ac友們cording to the 銀音conservativesequence of ly紅如mphocytic choriome問月ningitis virus（LCMV），and a real-time離飛 recombinase polyme生章rase amplification assay（real-time 內紙RPA）was established. Resu的明ltsshowed that the specificity of窗間 the assay was good，andno any高月 cross reaction with other murine 視去viruses detected by月遠 LCMV was shown. Thesensitivity of 文動established assay was as hi低黑gh as the RT-PCR method. Otherwise，bas國笑ed on detection of LCMV infected m新分urine tissues and negativeserums，the r還刀eal-time RPA show土通ed good stability 些西andreliability.土笑 Compared with current det低窗ection methods of nucl市刀eic acids，the test results coul件男d be interpreted within 20 minute事用s after loadingthe n書來ucleic buffer using th呢銀e real-time RPA，which 報好couldsatisfy the need of rapid LCMV 姐制inspection in roden件在t animals. 全文下載鏈接：這刀http://kns.cnki.net/KCMS/detail/37.1放白246.S.20190605.1615.042.html
2019-06-26
一株類禽型H1N1豬流感病毒的進(jìn)化分析與分子特征
2017年12月，上海市一養殖場飼養的豬出現咳嗽、呼吸困難、發(fā)快如熱及迅速轉歸等病症。將(jiāng)豬鼻拭子雨秒接種(zhǒng)雞胚進(jìn)行病毒分離下森，對(duì)血凝試驗陽性樣(yàng)品在SPF雞胚上進(jìn)地船一步純化、增殖，分離到1株豬流感病毒（A/swine/Shan費畫ghai/1205/2017）。采會討用RT-PCR對(duì)分離毒株進(jìn)行全基樂下因組擴增測序，利用DNAstar軟件，對(duì)測序基因片段進(jìn)議火行整個閱讀框的核苷酸序列同源性比對(duì)分析，并用MEGA6繪制遺妹空傳進(jìn)化樹并分析氨基酸位點。結果顯示：分離毒株爲H電嗎1N1亞型，其8個基因片段均屬于類禽輛他型H1N1進(jìn)化分支，沒(méi)有出現不同基因型流秒低感病毒片段之間的重組；分離株HA蛋白的裂解位點序列爲PSIQSR子的↓G，具有典型低緻病性流感病毒的分子特征綠器。本毒株的分離鑒定爲分析我國(guó)大陸地區的豬流感流行狀如個況和分子特征提供了參考。Phylog年我enetic Analysis on 好音a Strain of Avian-like H1N1 Subt線朋ypeSwine Influenza Virus and Deter答呢mination of Its Molecular Charac能裡teristicsIn December 2017，pigs in afa很視rm of Shanghai City appeared a serie姐水s of symptoms，includingcough，d在討yspnea and fever，and thediseased pi山家gs recovered rapidly. The線紙n the nasal swab samples collected問很 frompigs were inoculated into SPF ch家照icken embryos for virus 如銀isolation，after further p拍書urifying and proliferating the posit兒來ive samplesdetected by h坐路aemagglutination assay數放（HA），one strain o公好f swine influenza virus（A/妹上swine/Shanghai/1205/2017）was isolated.事視 The whole genome of the i為做solate was amplified ands吧匠equenced by RT-PCR，the nucleotide風高 sequence homology ofthe wh綠睡ole reading frame wa拍相s analyzed by the DNAstar software，and 國問then the genetic evolution tree 頻商was designed using影地 MEGA 6 toanalyze th畫呢e amino acid sites. The results showed 拍要that the isolated strain waside人知ntified as H1N1 subtype influenza 兵些virus，and itseight gene農短 fragments were classified into evolut金小ionary branch of avian-likeH1N1，w哥個ith no any recombination am懂睡ong influenza virusfragments呢光 of various genoty我錯pes. The cleavage site sequence of its 說也HA proteinwas PSIQ行通SR↓G，indicating the購視typical molecular characteristi短醫cs of low pathogenici農腦ty influenza vi舊討rus.Therefore，the isolati還歌on and identificati什睡on of H1N1strain 河動would provide some器北 reference for analyzing慢雪 the epidemic situ分金ation andmolecul下聽ar characteristics of swine 國低influenza in Mainland China. 全文下載鏈接：有場http://kns.cnki.net/KCMS/detail現森/37.1246.S.20190605.1615.038.html
2019-06-26
動物源食品中呋喃西林及其代謝物氨基脲研究進(jìn)展
呋喃西林是一種(zhǒng)硝基呋喃類抗菌藥物，曾被很那(bèi)廣泛應用于水産養殖過(gu公車ò)程中。呋喃西林在動物體内的代謝物氨基脲（SEM）會(huì)與細胞子頻膜蛋白緊密結合，形成(chéng)結合态殘留物，穩資術定而長(cháng)期存在于動物體内，具有緻癌、為他緻畸性。本文簡要介紹了呋喃西林藥物的藥理作用及其應用，綜述了兩(liǎng)種也金(zhǒng)應用于SEM檢測的主要方法——高效液相色譜（HPLC）及筆去其聯用技術和免疫分析法的研究進動愛(jìn)展情況及其優缺點，以及食品中的SEM來源，以期對(duì)呋喃西術外林藥物及其代謝物SEM有一個較爲完整的了解，也爲朋們進(jìn)一步開(kāi)展相關研究奠定理論基礎。司但Research Progress 從分on Nitrofurazone and Its Metabo坐中lite ofSemicarbazide in Animal-d下小erived Food Nitrofurazone，as anitrof議黑uran antibiotic，had been wid能讀ely used inaquaculture. However計站，the semicarbazide（SEM），a kind of坐輛 metabolite generated byn朋低itrofurazone within animal&現器#39;s body，could closelyincorpo朋離rate with cell membrane protein to for北化m bound residues which wo我樹uldexisted in animal's 討一body for a long time with carc醫海inogenicity andterat學紅ogenicity. In this pa什電per，the pharmacologicalef黑國fects and application of n話他itrofurazone were brief花呢ly introduced. Besides，the research pr行哥ogress，advantages anddisadvan鐘公tages of the main methods for 從煙SEM detection，i又西ncludinghigh performance li也少quid chromatography（HPLC）and its combi一弟ned technology and immunoassay were相土 summarized，the so物短urces of SEM in food was also我花 summarized，hoping to comprehensive呢玩ly recognize the nitrofur員妹azone and itsmetabolite of semicarba麗音zide，and to provide the自又oreticalfoundation for further research藍我. 全文下載鏈接：http://kns.cnki.net/舞土KCMS/detail/37.1246.S.20190605.村了1615.030.html
2019-06-26
中藥抗病毒性疫病作用機制研究進(jìn)展
病毒性疫病作爲一類具有高度傳染性的疾病，目前尚缺乏慢秒理想的抗病毒藥物，而中藥因其獨特的病毒工城病防治視角，成(chéng)爲近年來我國(guó)學(xué)者們研究的熱點中個。研究顯示，多種(zhǒng)中風票藥及其有效成(chéng)分對(duì)病毒有一定的抑制作用。本文結合病毒科坐性疫病發(fā)病機理與近年來抗病毒中藥研究成(chéng)果，金離對(duì)中藥的抗病毒機制進(jìn)行樂銀闡述，以期爲多方位、多靶點、天然低毒獸醫臨床抗病毒藥物的研發(fā)多快提供幫助。Research P了暗rogress on Functional Mechani窗數sm of Traditional 頻筆ChineseMedicine Against Viral Di兵輛seasesFor vital匠個 diseases with stro山白ng infectivity，there h村高as been still no any ideal antivira家開ldrugs，however，tradit城外ionalChinese me銀從dicine characterized by it件話s unique treatment perspective and me學校thodhas become a hotspot to be studied上有 by scholars in China. Res船時earches showed thatvirus c笑金ould be restrained to some機吧 extent by many Chinese medicines 章笑and theiractive in農錯gredients. In this paper，the mechan西大ism ofChinese medicine a討吧gainst viral dis影司eases was elabor我她ated in combination with thepathogen有男esis of viral diseases and th舞作e research results of antiviral Chin店吧esemedicine in recent yea水南rs，so as to provide basis todevelop mul放問ti-directional，multi-t下著arget and naturallow-toxic antivi算鄉ral drugs in veterinary practice. 又車全文下載鏈接：http://kns.cnki離遠.net/KCMS/detail/3山費7.1246.S.2019060區唱5.1615.028.html
2019-06-26
禽流感病毒實時(shí)熒光定量RT-PCR檢測方法的建立那相
爲滿足禽流感病毒高通量快速檢測的需要，建立了一種(zhǒng)能(né謝微ng)夠檢測各亞型禽流感病毒的實時(shí)熒光定量RT-窗什PCR檢測方法，并對(duì)該方法的特異性和靈敏度進(jìn)行評估，使去山用該方法與農業行業标準NY/T772—2013中的禽流河公感病毒RT-PCR方法同時(shí)進(jìn)行臨床樣(yàng)品檢測微說。結果顯示，該方法檢測耗時(shí)短、特異性好(hǎo)，檢測下限達10-雨長4 ng/μL，與傳統的RT-路討PCR方法陽性符合率爲100%。結果表明，本方法能(néng)村還實現對(duì)禽流感病毒的安全、特異、快速、靈敏、信上簡單、高通量檢測，從而彌補了現有傳統檢測技術的不足。 Establishm媽劇ent of Real-time RT-見歌PCR for Detecti習綠on of AvianInflu可東enza VirusIn order to achieve the志弟 high-throughput an空上d rapid detection ofavian infl答個uenza virus（AIV），自我a real-time fluorescent quan冷車titative RT-PCR method was establishe雪微dfor detecting all subtypes議海 of AIV. Then its specificity and sensi愛的tivity wereevaluated，and some clin文長ical samples were testedsimultaneously 地制by the established method and the 地南conventional RT-PCR 外民stipulatedin the agricult腦算ural industry standard N都微Y/T772—2013. Results showed t服妹hat thedetection limit o會月f the established method could reach用照 10-4 ng/μL with short綠腦detection time an和紙d better specificity，and the positiveco熱視incidence rate was 100就資% as compared with traditional RT水議-PCR method. Inconclusion，the esta說算blished real-time RT-PCR 呢技method wassafe，specific，rapid，sensitiv一聽e and simple，and could achi上校eve thehigh-throughput 商亮detection for AIV，which covered thedefi業購ciency of current traditional detection那術 technologies.全又慢文下載鏈接： http://kns.cnki器章.net/KCMS/detail/37.1246.S.20190422.101著線8.032.html
2019-05-10
萊姆病，離我們有多遠?
最近，“蟲蟲科學(xué)咖啡館”微信公衆号推送可身了一篇高強撰寫的萊姆病科普介紹文章。文章從萊姆病發(fā)現史、病原熱就學(xué)、傳播途徑、發(fā)病症狀、預防措數日施幾個方面(miàn)進(jìn)行了詳看視細介紹。萊姆病是1975年發(fā)現的一種(zhǒng)人獸共患自然疫舊子源性疾病，因發(fā)現地在美國(guó)康州的老萊姆鎮而被(bèi樹那)首次命名。1982年，Burgdorfe購對ri等首先從紐約長(cháng)島萊姆病疫區的肩突硬蜱中分離出該病病光現原體，後(hòu)被(bèi)命名爲伯氏疏螺旋體。該病目前在全球30兵紙多個國(guó)家和地區分布，主要分布在美國(guó)東北部吧月、中西部和西部，加拿大東南部以及我嗎歐洲中部及北部，亞洲東部和北非。我國(guó)于1986年首次在黑草工龍江省林海縣發(fā)現萊姆病，并于1鄉很990年從蜱中分離出病原體。目前經(藍訊jīng)流行病學(xué)調查及病原學(xué)證實，我國(guó)至少2服腦9個省(市、自治區)的山林地區人群中有科高萊姆病感染的存在，19個省(市、行花自治區)的山林地區爲萊姆病自然疫源地，13個省（市、自治區）有萊白在姆病散在發(fā)生和流行。該病的主要傳染源爲齧齒類動物，傳快都播媒介爲吸血蜱蟲，人群普遍易感。萊姆暗朋病臨床表現複雜多樣(yàng)，一般可分爲早、中、晚三期：關是早期以皮膚出現慢性遊走性紅斑（ECM）損害爲特醫生征，中期以心髒和神經(jīng)系統症狀爲主，晚期以關節炎和神經(jī音技ng)症狀爲主。通常在夏季和早秋發(fā)快議病，可發(fā)生于任何年齡的人群，男性略多于女性。發(fā)病以青影信壯年居多，與職業密切相關。以野外工作者、林業工人感染率較高。萊店兵姆病是蜱媒傳染病，其預防措施與其他蜱媒傳染病基本類似，最有效的預防藍是措施就(jiù)是接種(zhǒng)有效菌麗飛苗，但目前并未研發(fā)出有效的疫苗，因此目前視玩所能(néng)采取的主要預防措施就(jiù)是控制月事傳染源、傳播媒介、防治蜱叮咬。 全文下載鏈接： https:/資線/mp.weixin.qq.com/s/7X路多8RJ4B0nFwOgW9SVQxkdg
2019-05-10
鑒别H7N9流感病毒強毒和弱毒實時(shí)熒光RT-PCR方用師法的建立
爲建立可以區分H7N9流感病毒強毒風理株和弱毒株的方法，通過(guò)比對(duì)GenBan廠電k和GISAID中H7N9流感病毒的HA基因序列以及本實驗室保存的病毒序列草和，設計1對(duì)特異性引物和2條探針，建立了區分H7N9流感病毒強弱毒區森株的實時(shí)熒光RT-PCR術小方法，同時(shí)對(duì)該方法的反應體系和反應參廠工數進(jìn)行優化，并開(kāi事志)展特異性和敏感性試驗以及臨床應用試驗。特異性算冷試驗顯示，該方法具有良好(hǎo)的特異性，對(duì)其他常見亞型的禽流南麗感病毒和其他禽病病毒檢測均爲陰性。敏感性試驗顯店文示，該方法對(duì)H7N9流感強毒和弱毒開西HA基因的檢測下限分别爲0.004 fg 和0.1 fg RNA模闆，高醫國于兩(liǎng)種(zhǒng)商品化試工森劑盒的靈敏度。利用該方法對(duì)40株經(jī輛音ng)實驗室分離鑒定的H7N9流感病毒進(jìn)行對(du知來ì)比驗證，發(fā)現檢出率和符合率均爲100%。結果表明，該方法具新鄉有特異、敏感、準确的優點，可用于H7N9流感病毒的快速檢測，聽金同時(shí)還(hái)可區分強弱毒株，對(duì)H7N9流感病毒，尤坐自其是高緻病性流感病毒的早期診斷和有效防控具有重要意義歌那。 Development of a Real-time RT-PCR Met間你hodfor Identification 輛雨of Virulent and Avirulent S了服trains of H7N9Influ舞鄉enza VirusIn order to deve聽行lop a method for identif話服ying virulent andavirulent strains of花微 H7N9 influenza virus，a pair ofspecific又她 primers and probes wer花冷e designed by comparing the HA人科 gene sequences ofH7日是N9 influenza virus in為友 Genbank and GISAID and the sequence o又愛f which wasisol醫你ated and sequenced by our laborator近車y，a real-timeRT些愛-PCR method was developed，民她meantime，the reactio嗎章n system and paramet坐理ers of the method w唱短ere optimized，and the speci低信ficity and sensitiv廠多ity as well as its clinicalapplicati行資on were tested. 拍快The specificity test showed that，t地了he development method didn't r信農eact with other common s視美ubtypes ofavian influenza vir醫刀us and other avian vir做弟us，showing agood spec志問ificity. Sensitivity test sh關煙owed that，by theRT秒文-PCR method，the lower 看火detection limits 也自of HA genes ofvirulent and attenuated H人這7N9 strains were 0.004 fg and 0.去姐1 fg respectively，which wer錢兒e all lower than 風老the detection limits o吃喝f the two kinds ofcom電場mercial kits. By comparing and verifyin冷醫g 40 strains of H7N9 influenza vir妹技usisolated and identified 舊師by the laboratory，it wa民輛s foundthat both the detection rat問鄉e and coincidence rate were快湖 100%. Therefore, it wasconcluded th可了at the method could 哥空be used for rapid detecti請兵on of H7N9 virus andidentifyi都冷ng the virulent and avirule自海nt strains due to its advantages 也麗of goodspecificity，sensitivity and hi相地gh accuracy，which was of問工 great significance fo訊村r early diagnosis and是路 effectiveprevention and高數 control of H7N器文9 virus，especially thehi喝哥ghly pathogenic virus.全文下載鏈接視又： http://kns.cnki.net/KCMS/d地歌etail/detail.aspx?dbcode=CJFQ&dbna飛那me=CJFDPREP&filename=ZGDW201904015&資有v=MDA0NjZPZlkrUnFGeS9rVWJySlB5c就如lBlYkc0SDlqTXE0OUVZWVI4ZVgxTHV4WVM3年市RGgxVDNxVHJXTTFGckNVUkw=
2019-05-10
土拉杆菌病特征及其國(guó)内外流行狀況
本文簡要綜述了土拉杆菌病的病原學(xué)、流行病學(xué)、厭雜臨床症狀、病理變化等特征以及診斷方法和治療措施，介新知紹了該病在國(guó)内外的流行狀況。土拉南冷杆菌病在國(guó)外主要流行于北美和北歐地區。我國(什如guó)的土拉杆菌病自然疫源地主要集中在西藏地區，近年來人間報道(d市動ào)病例極少，動物土拉杆菌病僅有零星散發(fā)。我國(guó)應匠師對(duì)齧齒類經(jīng)濟動物和水貂等進(jìn)行病原學(xué)流拿書行病學(xué)調查，并對(duì)土拉杆菌有較強年說抵抗力的動物進(jìn)行血清流行病學(xué)調查喝雪，根據調查結果及時(shí)提出土拉杆菌病向(xiàng)人間傳播的預警。 些筆A Brief Introduction on Tularemia and亮費 Its Prevalence in多校 theWorldIn thi她話s paper，theetiology，epidemiology，嗎水clinicalsymptoms and pathological cha算姐nges of tularemia，as wellas the 電外diagnostic methods and treat著風ment measures were briefly summarized，b飛器esides，its prevalence 玩機both in and outsideChina was also 筆器introduced. Presently，tularemia ismai睡討nly prevalent in North America藍費 and North Europe. In China，i科美ts natural nidus is mainly conc光市entrated in Tibet. There have b問草eenvery few cas民厭es reported in human，and sporad和低ic outbreaksin animals. It was indica什一ted that etiologica大喝l and epidemiological in弟麗vestigationtowards rodent e文樹conomic animals and m動現ink，andsero-epi她花demiological investigation towards a影問nimals with strong resistance tot到鐵ularemia should be carried數麗 out in China，based on the外國research results，the early w雨西arning could be made intime.全爸唱文下載鏈接： http://k票火ns.cnki.net/KCMS線喝/detail/37.1246.S.20190422.1018.028.htm也明l
2019-05-10
牛結核病疫苗研究進(jìn)展
目前處于臨床試驗階段的醫用結核病疫苗共計15種(zhǒng)，其中吧相臨床I期6種(zhǒng)、Ⅱ期6種(zhǒng)房開、Ⅱb期2種(zhǒng)、Ⅲ期1種(zhǒng)。醫用疫苗厭腦的進(jìn)步爲牛結核病疫苗應用提供了更多的選擇。20世紀90年代起(紅離qǐ)，英國(guó)、新西蘭、西班牙刀來等國(guó)陸續開(kāi)展了野生動物的牛結核病免疫田間試驗。現在北訊全球雖然仍禁止使用疫苗免疫牛，但很畫店多學(xué)者仍然進(jìn)行了大量的店店牛結核病疫苗研究。英國(guó公但)動植物健康署（APHA）通過(guò)試驗證實，卡介公好苗（BCG）-病毒載體疫苗聯合免疫可強化BCG免疫效果。歐盟開(kā區城i)展了結核病階梯項目（TBSTEP）河聽，研究根除牛結核病的策略，其中包括疫苗和診斷試劑研發(f雨外ā)。此外，爲擴大試驗範圍，歐盟食品安全局還(hái)要求開城短(kāi)展牛結核病疫苗田間試驗。 Research司湖 Progress on Bovi得司ne Tuberculosis Va訊行ccinesPresently，a total村計 of15 kinds of medical tuber內銀culosis vaccines are at熱知 the stage of clinical trials，inc用什luding 6 kinds of vaccines at cli章討nical stage I，6 kinds at stage子訊 II，2 kinds at stage I問低I b，and 1 kind at stage III. With the木購 development of風路 medical vaccines，大得the bovine tuberculosis vacci師科nes would be applied in a lar器明ge range.Since the 19劇為90s，the field trial鄉技s for immunizationag的房ainst bovine tuberculosis 黑我in wild animals have been內電 carried out in th房大e UnitedKingdom，New Ze年熱aland，Spain andother countries. Altho民間ugh it is still prohibited to小明 use vaccines in ca一喝ttle inthe world，a large number o煙就f related vaccine researcheswere c上上arried out by many scholars. The immu煙志ne effects of bacil匠看luscalmette-guerin（BCG）could bestrengt亮章hened by combined immu紅錢nization of BCG器媽-viral vector vaccine as tested 通筆andconfirmed by B術影ritish Animal and Plant Healt樂作h Administration（AP照喝HA）. The Tubercu畫裡losis Staging Project（TBSTEP）w看遠as performed by European U問見nion tostudy strategies for eradicat又服ing bovine tuberculosis，includingt筆場he development of vaccines and外呢 diagnostic reag業話ents. In addition，th紙低e field trials for bovine tuber話紙culosis vaccines were說理 also requiredt子藍o carry out by European Union Food S電懂afety Authority so as to en機又large the scopeof trial媽謝s. 全文下載鏈接： http://kns.cnki.net讀新/KCMS/detail/detail討生.aspx?dbcode=CJFQ&d白司bname=CJFDPREP&filename=ZGDW又從201904013&v=MjI4的匠MDF5clBlYkc0SDlqTXE0OUVaNFI4ZVgx場村THV4WVM3RGgxVDNx鄉吃VHJXTTFGckNVUkxPZ對雜lkrUnFGeXpoVjdyQlA=
2019-05-10
牛結節性皮膚病預警信息與風險管理
近年來牛結節性皮膚病（LSD）已擴友冷散至俄羅斯和哈薩克斯坦，有可能(néng)在數月黑來或數年内傳入中國(guó)，從而對(duì)中國(guó水相)牛群帶來嚴重威脅。本文闡述了LSD在全球擴散情況、預警信息、流那日行特征、診斷方法，以及相應的風險管理措施。分析了保加利亞LSD有樹防控經(jīng)驗，包括：政府和科學(x木村ué)界高度重視，基于傳播風險調整防控措施，抓好(hǎo)感染短吧牛群的快速撲殺和無害化處理，大規模開(高和kāi)展疫苗免疫，限制活牛運輸等。 體數Early Warning Informa公作tion andRisk Mana筆少gement of Lumpy Skin 章低DiseaseIn recent years，lumpy skin厭師disease（LSD）has spread 門路toRussia and Kazakstan，and做農 is likely to be released i生又ntoChina in several months or都數 years，which may br妹站ing hugethreat to但算 the cattle in China. In this paper，校頻the spreadof LSD in t外林he world，early warn藍門ing information日制，epidemiological features，門視diagnostic methodsand releva事說nt risk management were e河哥xpounded. In addition，家從the experiences lear姐窗ned from the prevention and con城朋trol of LSD inBulgaria厭雪，including the high priority 道草given bygovernment and scientific circ這問les，were analyzed.Therefore，it was sugg為日ested that the risk of diseasespreadin南兵g should be time醫水ly followed by control measures，and the可行 core measures，such as rapid cull技學ingand bio-safety disposal 亮們of infected cattle，massvacc中科ination and restriction on liv們匠e cattle transpo化問rtation should be adopted andsupervi刀就sed by government. 全看體文下載鏈接： http:/頻內/kns.cnki.net/KCMS/detai是些l/detail.aspx?dbcode=CJFQ&dbname=CJFDP學門REP&filename=ZGDW201904012&v=M黃睡Tc1NDdTN0RoMVQzcVRyV0林內0xRnJDVVJMT2ZZK1JxRnl6aFVydkpQeXJQZWJHN紙數Eg5ak1xNDlFWm9SOGVYMUx1eF熱子k=
2019-05-10
H5亞型禽流感病毒實時(shí)熒光RT-PCR方法的建立與應用
H5亞型禽流感病毒HA序列差異越來越大。爲更準确地檢測H5亞型禽流東購感病毒，對(duì)GenBank中發(fā)表的H5亞型禽流感些物病毒HA序列以及本實驗室保存病毒序列進(jìn)行比對(duì)了爸，同時(shí)設計1對(duì)引物和1條探針，建立了H5亞型禽流感病機舞毒實時(shí)RT-PCR方法，并對(duì)該方法的反應體系醫線和反應參數進(jìn)行了優化。以本實驗室分離輛電測序确定的30份H5亞型禽流感病毒子放RNA爲模闆，將(jiāng)該方法與兩(li服訊ǎng)種(zhǒng)商品化H5亞型禽流感病毒檢測試劑盒進(jìn的動)行比對(duì)，發(fā)現該方法與兩(草相liǎng)種(zhǒng)商品化試劑盒的器照檢出率分别爲100%、98%、98%。敏感性試驗顯示，該方法可以檢舊南測出0.1 fg的RNA模闆，靈敏度比兩(liǎn舞白g)種(zhǒng)商品化試劑盒均提高了10答用倍。結果表明，該方法具有更高的特異性和敏感性，如店可用于H5亞型高緻病性禽流感的早期診斷。她錯Establishment and有懂 Application of Re嗎事al-time RT-PCR for Detectionof H5 Subt熱照ype Avian Influenza VirusIn re吃他cent years，thedifference商開 in HA sequence of H5 subtype avia來文n influenza virus（A草拿IV）was increasing. In order to de錯說tect H5subtype AIV more accurately，by分用 comparing the HAsequence of H5 愛森subtype AIV published 路離in GenBank with the one conserved in t謝動helaboratory and design影到ing a pair of primers and one Ta分下qMan probe，a real-time RT分她-PCR method was developed，andit大們s reaction system and param林服eters were optimized. Taking 3著書0 RNA samples of H5subtyp商笑e AIV strain isolated and se外很quenced by the labora習去tory as the template，the establ算上ished method was compared with tw票鄉o commercial H5 subtypeAIV detecti器腦on kits，and it was found能厭 that the detectionra討有tes were 100%，9線但8% and 98% respectively. Sens現朋itivitytest showed that RNA t火拿emplate of 0.1 fg co們哥uld be detected by the establis飛讀hedmethod whose sen你了sitivity was 10 times higher than 國那that of the two commercialkits. In co車年nclusion，the RT-PCR me化銀thod was more specific我了and sensitive，and i業遠t could be used f門照or early diagnosisof H5 subtype hig近坐hly pathogenic avian influenza. 全文下長科載鏈接： http://kns.cnki.net/KCMS/deta拿窗il/detail.aspx?dbcode=CJF銀看Q&dbname=CJFDTEMP&filename=ZGD校做W201903020&v=MDczNzBScUZ5emdV呢嗎YjdNUHlyUGViRzRIOWpNckk5SFpJUj雨玩hlWDFMdXhZUzdEaDFUM3FUcldN鐵木MUZyQ1VSTE9mWSs=
2019-05-10
藍舌病I型病毒VP5蛋白的表達純化及免疫原性分析
爲分析藍舌病I型病毒（BTV-1）VP5蛋白的免疫原性，本研究構建了載體P志什ET-28a-VP5并轉化BL21（DE3），然後(hòu)進(理跳jìn)行IPTG誘導表達，以及SDS-PAGE和Western-b術放lot分析。結果顯示，重組VP5蛋白相對(duì)分子質量兒會約爲62 kDa，與預期結果一緻。進(jìn)一步優化條件，發(fā)現鐘朋IPTG爲0.2 mmol/L、溫度爲25℃、誘導6 h到老時(shí)，重組蛋白在上清中表達量最高，通過(guò家知)Ni柱純化獲得的重組蛋白純度約爲75%。將(jiāng)看件純化後(hòu)的重組蛋白VP5分别以40μg/隻和20呢請 μg/隻免疫Balb/c小鼠，同時(shí)設置PBS爲陰性對(d少市uì)照，對(duì)三免後(hòu)血清進(jìn)金歌行間接ELISA檢測，發(fā)現高、報女低劑量免疫小鼠均能(néng)産生針對(duì)VP5蛋化路白的特異性抗體。用滅活BTV-1進(jìn)行東懂DOT-ELISA檢測，發(fā)現其暗姐能(néng)與VP5蛋白免疫的小鼠血清發(fā)生特異性反應，說(sh黑長uō)明利用大腸杆菌表達的重組蛋白VP5具有良好(hǎo)的免疫特性，大商這(zhè)爲進(jìn)一步開(kāi)展BTV相關研究奠定了基礎。 E房來xpression，Purification a數區nd Immunogenicity Analysis onVP5要員 Proteinof Bluetongue Virus Serotype吧草 IIn order to analyze the 章信immunogenicity of VP5 protein of blue畫著tonguevirus serotype I（BTV-1），in this p吧謝aper，the prokary行放otic expression vec森劇tor PET-28a-VP5畫喝 was constructed andtra北呢nsformed into BL21（DE3）.After inducible藍房 expression by IPTG購近，the VP5 protei對亮n waspurified and analyzed年弟 by means of SDS-PAGE and Western-blo湖從t. The results showedthat 吃資the relative mo兒村lecular mass of reco電姐mbinant VP5 protein was about 62 k小船Da，which was consistent with the e黃票xpected result. It was 民作found that，by further opti那內mizing，the expression l紅吃evelof recombinant protei不厭n in supernatant was maximum w了化hen IPTG was 0.2 mmol/L，the bacteria s討技olution was inducted for 6 hours嗎購 at the temperature of25℃，and the書要 purity of recombinant VP5 p國黑rotein throughNi-se如理pharose purification was about 7舞醫5%. Then the puri友廠fied VP5 protein wasimmunized to Bal去西b/c mice at the doses of 4音線0 and 20 μgper mo快電use respectively，meantime，PBS was 了通set as the negative control. The serum多但 samples were testedb雜雪y indirect ELISA after being im開雪munized for 3 times. It was conclude村黑d that thespecific antibody again新說st VP5 protein appeared 會湖in immunized mi木內ce with high orlow dose. In addit遠的ion，it was found that inac為人tivatedBTV-1 could specifical就河ly react with the seru哥行m of immunized mice with V物裡P5protein by DOT-ELISA test，ind年煙icating that theimmunogenicity of recom拍腦binant VP5 protein was good，whichl愛拿aid the foundation for furt話唱her research on BTV. 全文下載鏈接： h湖答ttp://kns.cnki.net好月/KCMS/detail/detail.aspx?dbcode=CJF新和Q&dbname=CJFDTE高房MP&filename=ZGDW201903016&v=MDg3N友暗TFxRnl6bldydkFQeXJQZWJHNE冷議g5ak1ySTlFWW9SOGVYMUx1eFlTN0體唱RoMVQzcVRyV00xRnJDVVJMT2ZZK1I=
2019-05-10
藍舌病I型滅活疫苗免疫綿羊後(hòu)中亮知和試驗和cELISA檢出抗體的特爸筆點與關系
爲比較藍舌病病毒（BTV）cELISA和血清中和草輛試驗（SNT）檢測方法的特點與關系，通過(guò)滅件答活、濃縮，制備抗原含量爲1、5、10、50、100 μg/mL的BTV醫紙-1型滅活疫苗。每個含量爲1組，并近知設對(duì)照組，每組6隻綿羊，分别在0、3周免疫。每周采血1次，共靜見采血6次。對(duì)所采血樣(yàng)分别現術用cELISA和SNT方法進(jìn)行抗體檢測。cE市熱LISA檢測結果顯示，免疫後(hòu)第1周，抗吃日體陽性率達到80.00%，第4周全部免疫羊轉爲抗體陽性，抗體産生較爲迅速；SN要子T檢測結果顯示，中和抗體陽性率從免疫後(hòu)第1周的6.67了黃%持續上升至第3周的36.67飛線%，至第4周達到70.00%，中和抗體産生持續而緩慢。随著(z靜對he)時(shí)間延長(cháng)，2鐵票種(zhǒng)檢測方法的符合率由來朋23.33%增加到76.67%。當能金綿羊同時(shí)産生中和抗體和cELISA抗體時(shí)，中和抗體效價錯這與cELISA抑制率間有顯著的線性相關關系（r=0.63）習多，相關方程爲y=0.38+0.05χ。分析認爲，中和抗體出現的時木空(shí)間之所以晚于cELISA抗體，可能(néng)是因爲自資與宿主細胞識别并吸附的主要部位VP7能們蛋白位于病毒衣殼的中間位置，其主要表面(miàn)抗原位點暴露需要時(sh也路í)間。 Characteristics an這多dRelationship between cELISA Antibo謝做dy and Neutralizing AntibodyProduce站船d by Stimulating the Sheep with BTV-1如國 Inactivated VaccineIn order to compare鐵年 the characteristics and explore 又務therelationships between cELISA and the影市 serum neutralization test（SNT）in de說算tectingbluetongue virus（BTV），B筆劇TV-1inactivated vaccines were prep用飛ared byinactivating and concentrating，校女and there were fivekinds of concentra和熱tions：1，5，10，50 and 100 μg/mL，respe這很ctively. Each ki黑厭nd of concentration was considered as飛雨 one groupconsisting of 6 sheep whic畫紅h were immunized in the 0th a商生nd 3rd weekrespectivel窗鐘y. Also，the control內和 group was set. Bloodsample舞現s were collected once a 子草week for 6 times，筆鄉whichwere tested by cE章路LISA and SNT res城志pectively. The cELISA resul大上ts showed that thepo兵木sitive rate of antib海為ody reached 80.00% in the first week 我紅after vaccination，and turn近玩ed to be 100% in the four畫微th week，showingthat the antibod討放ies were produced relativel技畫y rapid. While the 歌都results by SNTshowed that the positiv綠靜e rate of neutraliz秒拍ing antibody increased 公子from 6.67% inthe first week to 白民36.67% in the third 她男week after vaccination，and reached 70.內購00% in the fourth we見睡ek，indicatingthat the neutralizing ant的時ibody appeared continu生舊ously and slowly. Th暗西ecoincidence rate of the two me自新thods increased from 23.33% to 水鄉76.67% over time.When the neut紅黑ralizing antibody and影弟 cELISA antibody appe個很ared in sheep at thesame time，the頻愛re was an obvious linea輛鄉r correlation（r=0.63）occurred between 草影the neutralizingantibody tit唱件er and the inhibition道業 rate of cELISA，andthe correlation 友工equation was y=0.38+0.05χ. It wasa兵兵nalyzed that，the reason了報 why the neutralizing 樹好antibodyappeare兵林d later than cELISA antibo機雨dy was because VP7 protein rec慢吃ognized andadsorbed by host cel道頻ls was located 和這in the middle of醫身 viral capsid，thus it would 書對take some time to expose its 農作main surface anti年飛gensites. 全文下載鏈接： http://kn區懂s.cnki.net/KCMS知下/detail/detail.aspx?dbcode刀暗=CJFQ&dbname=CJFDTEMP&filenam章為e=ZGDW201903015&v=MzI3Njc0SDl近市qTXJJOUVZWVI4ZVgxTHV行現4WVM3RGgxVDNxVHJXTTFGck購西NVUkxPZlkrUnFGeXpuVjdyTlB5clBlYkc=
2019-05-10
2017—2018年雲南省邊境地區阿卡斑病毒抗體檢測來和
爲了解雲南邊境地區近年來阿卡斑病毒（Akabanne disease viru答朋s，AKAV）流行情況，2017—2018年連續2年在綠時雲南省與緬甸、老撾、越南接壤的12個邊境通弟縣采集牛血清，應用cELISA方法雜看進(jìn)行AKAV抗體檢測。結果顯示：2017—2018年共檢測牛血現亮清樣(yàng)品1860份，檢出陽性樣(yàng)品108份，陽性率爲長拿5.8%，顯示這(zhè)些地區影報存在一定的AKAV感染；各縣AKAV抗體陽性率在0~24.4%之呢少間，差異較大，其中抗體陽性率最高的爲富甯縣，哥海2017年和2018年的陽性率分别爲24.4%和20.0%，而瑞樂動麗連續2年未檢出抗體陽性；黃牛AKA森去V抗體陽性率（6.6%）高于水牛（4.7%）西綠，差異顯著（P
2019-05-10
洛克沙胂完全抗原的制備及其單克話中隆抗體的研制
本研究以洛克沙胂（ROX）的結構類似物3-氨基-4-羟基苯胂酸（HA我音PA）爲小分子半抗原，采用碳二亞胺法，分别將(jiāng什音)其與載體蛋白——牛血清白蛋白（BSA）少讀和雞卵清白蛋白（OVA）偶聯，制備免疫抗原HAPA-BSA和檢測抗原商腦HAPA-OVA。經(jīng)紫外光譜掃描及聚丙烯酰胺凝膠電泳（S跳器DS-PAGE）鑒定後(hòu)下海，以20 μg/隻劑量的HAPA-BSA免疫BALB/c小鼠慢機；選擇血清效價高、敏感性強的小草技鼠，取脾髒進(jìn)行細胞融合，制備雜美爸交瘤細胞株。利用間接ELISA山離法篩選得到1株可以穩定分泌抗ROX單克隆抗體的細胞株市雜，命名爲G12；間接ELISA法測定其細胞上清效價爲1:512，窗物間接競争ELISA 測定其半數抑制濃度 I河熱C50爲3.147 ng/μL。經(jīng)小鼠體内誘生腹腦鄉水，辛酸-硫酸铵法純化獲得純度身作較高的單克隆抗體，間接ELISA測定其效價爲1:102 400，IC50爲1路吃.433 ng/μL，且具有良好(上微hǎo)的特異性。本試驗成(chéng)功員木合成(chéng)了ROX人工抗原，并制備了可以分泌高敏感好鐵性單克隆抗體的雜交瘤細胞株，爲ROX免疫學(xué)視讀快速檢測方法的建立奠定了基礎。Syn校爸thesis of RoxarsoneCo討中mplete Antigensand Prepa章師ration ofCorrespondi風南ng Monoclonal Antibodie吧著sIn this research，3-amino-4自冷-hydroxybenzoic acid（HAPA），a st微白ructural analogu但坐e of Roxarsone（ROX）was鄉微 adopted as the small molecul雨信e hapten. By beingconjugated respectiv飛上ely with carrier protein albu男聽min-bovine serum（BSA和雜）and chicken ovalbumin（CVA）through carb媽腦odiimide method，the immune an大冷tigen HAPA-BSA and detection antig窗友enHAPA-OVA were produced. 友河After UV spectroscopy scan and p購遠olyacrylamide gelelectrophoresis（SDS雜體-PAGE）verification，HAPA-BSA dosage of 2場遠0 μg/per mouse was immunized to BA飛筆LB/c mice，then the 為吧spleens of mice w關那ith higher serumtiter and sensitiv玩外ity were taken for cell fus會紅ion to prepare hybridoma ce南暗llstrain. After screening嗎通 by indirect ELISA醫吧 method，one cell strain na務紙med G12 stably secreti話日ng anti-ROXmonoclonal antibody was obta物哥ined，and the titer in cell supernatant 鄉紅was 1:512，the inhibitory co少近ncentration（IC50）was measured家煙 as 3.147 ng/μL. Furthermore，t技讀he ascites produced in abd相站ominal cavity ofmice wer時藍e purified by caprylic輛熱 acid-ammonium sulfate method，and h可技igher purity of Abs were obtained，the t喝子iter of which was 1:102 400，and IC50 w風照as 1.433 ng/μL an公中d great specificity was sho民男wed. Inconclusion，ROX artificial antige爸又ns were successivelysyn兒東thesized and the h媽拍ybridoma cell line報中 that could secret high sensitivemonocl船書onal antibody was prepared，which wo機間uld built a foundation t謝老o establish ROX 美舊rapidimmunological detection m姐低ethod. 全文下載鏈接：htt妹大p://kns.cnki.net/kcms/detail/37.124白火6.S.20190222.0911.046.html?ui師什d=WEEvREcwSlJHSldTTEYzU3Eyd場很DRPZTFBeXhDa29ZZ2ljVUtvTXF2T計章3dtND0=$9A4hF_YAuvQ5對關obgVAqNKPCYcEjKensW4IQMovwHtwkF4VYPoHbK北弟xJw!!
2019-03-04
MALDI-TOF-MS用于微生物時有鑒定的影響因素分析
影響MALDI-TOF-MS微生物鑒定的因素有很多。本研究主要從微生物培養草農基、培養狀态、菌體前處理、上樣(yàng)量等方面(很信miàn)進(jìn)行MALDI-TOF-MS操作技術和方法的探讨。研究結果鐘子顯示：進(jìn)行MALDI-TOF-MS細菌鑒定，培術暗養時(shí)間在4~24 h之間爲佳；盡量選少麗擇無色、不含鹽的液體培養基；0.1 μL菌液是最低鑒定菌量；上樣(yàng)量算和對(duì)結果無影響；對(duì)于菌體前處理，除按照标準化前處理程序操作放拿外，也可將(jiāng)菌體加ddH2O制成(chéng費火)混懸液進(jìn)行點靶鑒定，這(zhè)樣(yàng)更加簡便。本研就技究有助于減少MALDI-TOF什我-MS操作盲目性，提高鑒定質量，利于操作的規範化和簡便化。Influe票的nce Factors Analysis onM火人ALDI-TOF-MS in Microorganism Identifi中下cationVarious factors could 裡街influence the ef計拿fectof MALDI-TOF-MS in microorgani金樂sm identification. In 海不this study，the approach去聽es and techniques ofMALDI-TOF-MS fr生也om aspects of microbiological medi樂雨um，bacteria incubation stat白姐e，bacteria pre-treatment and sampl紅到e loadingquantity北火 were summarized a著通nd discussed. According to 商我the research results，the period of b能音acterium incubation又白 of 4 to24 hours 會開was the best；colorless能資 and salt-free medium was pre嗎說ferable；the lowest as要票certainable bacter著老ia solutionwas 鐵草0.1 μL；the sample loading quan匠間tity had no effect onidentification r厭會esult. Besides，for the standard 多鐘pre-treatment p吧但rocedures，a han理男dier way was to a人聽dd ddH2O into t腦文he bacteriato make suspension and c秒頻onduct targetidentificat道但ion. This study would contribute to商樹 reduce the operational blindne短新ssof MALDI-TOF-MS，e樂頻nhance the identificat些綠ion quality andfacilitate the standard來靜ization and simplification of operation麗路. 全文下載鏈接：http://kns.cnki.舞冷net/kcms/detail/37.1246.S.20190222.091自近1.044.html?uid=WEEvREc北風wSlJHSldTTEYzU3EydDRPZTFBeXhDa29河也ZZ2ljVUtvTXF2T3dtND0美為=$9A4hF_YAuvQ5obgVAqNKPCYcEjKensW4動票IQMovwHtwkF4VYPoHbKxJw!!
2019-03-04
犬副流感病毒D121004株的分離鑒湖東定及其F基因遺傳進(jìn)化分析
爲了給犬副流感病毒（CPIV）疫風下苗研制提供支持，無菌采集有呼吸道(dào)症狀病死犬的腦和肺髒樣(又地yàng)品，處理後(hòu)接種(zhǒng)Marc-1低場45細胞，通過(guò)RT-PCR、紅細胞吸附試驗，對(duì)盲傳3代場金的細胞培養液進(jìn)行鑒定，成(chéng地身)功分離到1株CPIV。該病毒能(néng)使Marc-145細是電胞出現細胞病變，且具有吸附雞紅細胞的能(néng農懂)力，將(jiāng)其命名爲CPIV-D1在匠21004株。對(duì)該毒株的F糖蛋白全一海基因進(jìn)行克隆及核苷酸同源性分體些析并構建遺傳進(jìn)化樹，發(fā)現分離株的會少F基因較爲保守，未出現大的遺傳變異。今後(hòu)需要對(d但線uì)分離株的免疫原性進(jìn)行研究，以探究其理弟作爲候選疫苗株的潛力。Isolation and Identificatio遠東nof Canine Parainfl是離uenza Virus D121004 Strainand 你筆Sequence Analysi坐鐘s on ItsF GeneIn orde術森r to research and 還紅develop a vaccineagainst can女不ine parainfluenza 去離virus（CPIV），the brains and lungs o腦文f dead dogs suspiciou湖報s of respiratory sy微工mptomswere collected aseptically，and th農生en Marc-145 cells were inoculated after又黑 treatment to iso家見lateCPIV. RT-PCR and erythrocyte a一跳dsorption assay we議事re applied to identify the cellculture 呢要medium with three blind pas電影sages. The results showed that one st歌間rain ofcanine parainf器店luenza virus，CPIV-D1210書好04，was successfully isolated，which coul到喝d cause cytopathic ch通地anges in Marc-1音姐45 cellsand was capable of adsorbing ch內工icken erythrocytes. Cl視通oning and nucleotide h快視omologyanalysis on the whole F glycop聽我rotein gene were carried out，and th計山e genetic evolution tree was bu月間ilt.The results showed that the 花見F gene of the isolated strain was re光樹lativelyconservative，indicating no maj哥我or genetic variation. Infutu大都re，the immunogenicity of t廠視he isolate should befurther男山 studied，so as to explo科頻re the potential to be t爸多hevaccine strain. 全文下載鏈接：ht區這tp://kns.cnki.net/kcms/deta訊線il/37.1246.S.20190222.0了笑911.040.html?uid=WEE數人vREcwSlJHSldTTEYzU3EydDRPZTFBeXhDa29ZZ2議化ljVUtvTXF2T3dtND0=$9A4hF_Y生輛AuvQ5obgVAqNKPC喝會YcEjKensW4IQMovwHtwkF4VYPoH區數bKxJw!!
2019-03-04
沙門氏菌LAMP可視化檢測方法的建立
爲實現沙門氏菌的快速現場檢測，根據沙門氏菌 invA 基因編碼區序列，設計合成睡能(chéng) 1 套引物，通過(guò她紅)對(duì)反應物濃度和反應條件優化，建立了沙門氏菌屬特異性 LAM事術P 檢測方法；通過(guò)對(duì) 去劇invA 基因 8 個區域的 笑睡6 條引物配對(duì)進(jìn)行等溫擴增，并在反應體系中微看加入 HNB 來指示反應結果，實現了反應的快速閉管檢測。靈敏度試驗顯示，建立明時的方法可以檢出稀釋至 3.6×101 CFU/mL 菌液所提取的到我 DNA；特異性試驗顯示，建立的方法與大腸杆菌 DNA、志賀氏菌 DNA事我、布魯氏菌試管凝集抗原 DNA、炭疽坐愛沉澱抗原 DNA 以及豬鏈球菌 2 型 DNA 不發(fā)生交叉空兒反應，但能(néng)檢出雞白視微痢、禽傷寒沙門氏菌凝集抗原以及馬流産試管凝集抗原提取的 DNA；重報資複性試驗顯示，3 個稀釋度菌液 DNA 的 3 次重複檢測均爲陽厭公性；應用試驗顯示，從經(jīng)增菌培養的 2學場00 份進(jìn)境猴糞拭子樣(yàng)品中檢出陽性 2 份，與細菌分離坐路鑒定檢測結果一緻。結果表明，所建立的方法敏感性和動訊特異性較高，重複性滿足實際要求，可用于進(jìn)境動物樣(yà場兒ng)品的沙門氏菌檢測。Development of a Visual司錢 LAMPTest Method for事場 SamonellaIn searching of 黃是rapid field detection m路分ethods forsalmonella，a set of prime自短rs were designed船聽 andsynthesized–which were 3 文事pairs of primers thatidentify算紅ing 8 distinct regions場是 of invA gene of Salmonella for 國煙isothermalamplification. With optim子匠izing the reactant concentration and re笑制actionconditions，and by adding HNB 拿開to the reaction 鄉拍as anindicator，a closed-t中看ube LAMP method fo音暗r rapid fielddetection of Salmon司村ella was developed. Sensitivity test影火 showed that the DNAsextra舞坐cted from 3.6×101 CFU/mL bacteri船日al suspension could be detecte工城d. Specificity test木美confirmed that no 風放cross reaction was found between th西器e method with the DNAs ofGoliba們知cillus，Shigella，Brucella agglutinatin訊土g antigen，anthrax厭房 precipitation antigen 爸朋and type 2Streptococ草刀cus suis，but it could detect DNAs from 妹場pullorosis，Salm音麗onella typhi aggluti內請nating antigen andAbortus-e舞慢qui agglutination暗睡 antigen. Repetitiv大月e test verified that the re下來sultsof 3 experiments for DNAs 麗雜in 3 diluted bacteria solutions w快公ere all positive.Application 我哥test indicated that 2 of 20場媽0 swab samples from manures of新土 importedmonkeys were found positive關有，which was identical with the result票紙 of bacterial isolation test. 森從Alltest results proved that the m作訊ethod is featured with high新雜er degree ofsensitivity 海和and specificity，and it睡不s repeatability allows practical app他們lication，hence it would be a bette就習r approach todetect Salmonella子鐵 in animal import quarantine. 全文下載鏈接：ht年費tp://kns.cnki.ne她一t/kcms/detail/37.1246.S.20190222.0911.0科公36.html?uid=WEEvREcwSlJHSldT子學TEYzU3EydDRPZTFBeX白老hDa29ZZ2ljVUtvTXF2T3dtND0=$9A4月長hF_YAuvQ5obgVAqNKPCYcEjKensW4IQM議小ovwHtwkF4VYPoHbKxJw!!
2019-03-04
農業農村部辦公廳關于開(kāi)展2019年省級獸醫系統實驗室檢測能(né唱身ng)力比對(duì)工作的通知
農業農村部辦公廳關于開(kāi)展2019年全國(guó)高級别動她哥物病原微生物實驗室生物安全專項檢查工作的通知
韓長(cháng)賦會(huì)見世界動物衛生組織總幹事(shì)艾略特和聯行路合國(guó)糧農組織助理總幹事(s船得hì)提詹尼
農業農村部辦公廳關于印發(fā)《2場很019年國(guó)家動物疫病監測與流行病學間森(xué)調查計劃》的通知
國(guó)際布魯氏菌病專家Jacques Godfroid 教授中歌訪問我中心
我中心獲得2019年度山東省農牧微劇漁業豐收獎成(chéng)果獎二等獎
關于在加工流通環節開(kāi)展非洲豬瘟病毒檢測的公告
全國(guó)外來動物疫病綜合防控技術培訓班在青島舉辦
農業農村部七項措施穩定生豬生産
王功民指導中心2020年度動物衛生風險評估及業街國(guó)際獸醫事(shì)務工作
