豬源冠狀病毒監測簡報
目前，已知感染豬群的冠狀病毒有6種(zhǒng)志海，分别是豬流行性腹瀉病毒（PEDV）、豬傳染性胃腸炎病毒男理（TGEV）、豬呼吸道(dào)冠狀病毒（PRCV）、豬急性吧分腹瀉綜合征冠狀病毒（SADS-CoV）、豬血凝性腦脊髓炎病毒（PHEV）和快秒豬δ冠狀病毒（PDCoV）。在冠狀病毒科α、β、γ、δ等4個屬中，PEDV、司水TGEV、PRCV、SADS-CoV均爲愛草α冠狀病毒，PHEV爲β冠狀病毒，PDCoV爲δ冠狀病毒。PRCV大地在臨床上引發(fā)仔豬呼吸道(dào)症狀，PHEV感染會(huì)姐我造成(chéng)仔豬嘔吐和神經(jīng樹刀)症狀，其他4種(zhǒng)病毒均可引起(qǐ)豬群腹鄉個瀉。北京市延慶區農業局（北京市延慶區動物衛生監督管理局）對(duì坐秒)2011年以來收集自全國(guó)1就睡6個省份的2 915份臨床腹瀉豬群樣(yàng)品（腸道(dào)、糞便等）多站進(jìn)行了PEDV、TGEV檢測，檢出PEDV陽性1 223份、TGE數議V陽性109份，PEDV、TGEV檢出率分别爲41.96%、3.74%；對子地(duì)2014年以來收集自14個省份的12 43機長0份臨床健康豬組織樣(yàng)的船品（淋巴結、扁桃體等）進(jìn)行了PEDV檢測，檢黃短出陽性325份，檢出率爲2.61%。針對(duì)引發(fā)新型冠狀病毒肺炎路上的病原2019新型冠狀病毒（2019-nCoV），對(文厭duì)2018—2019年收集自15個省份的2 658份生豬雜公組織樣(yàng)品開(kāi一來)展了追溯性檢測，結果均爲陰性。對(duì)2019-nCoV與豬源冠狀病毒的國計親緣關系進(jìn)行了分析。通雪基于全基因組的遺傳進(jìn)化分析表明：2019-nCoV與同爲β冠狀病分離毒屬的PHEV核苷酸同源性僅爲54%，與α、δ屬豬源冠狀病毒親緣關廠舊系更遠。基于以上數據與分析，可初步排除2019-放近nCoV來源于已知豬源冠狀病毒的可能(néng)性Brief Repo明數rt on the Surveillance of Swine-orig為他in CoronavirusesCurrently，there鄉雜 are six kinds of coronaviruses known 請離to infect pigs，namely porcine epidemi問我c diarrhea virus（PEDV），東地transmissible gastroen農船teritis virus（TGEV），porc紙來ine respiratory coronavirus（PRCV），空到swine acute diarrhoea習藍 syndrome coronavirus（SADS-CoV），p業器orcine hemagglutinating enc短裡ephalomyelitis virus（PHEV）行低，porcine deltacoronavi睡路rus（PDCoV）. Among the four gen站動era of the family Coronavirid兵輛ae，PEDV，TGEV，PRCV and S如雨ADS-CoV belong to Alphacoronaviru雜時s，PHEV belongs to Betacoronaviru女年s，PDCoV belongs to Deltacoronavir月姐us. Clinically，PRCV cause熱吧s respiratory symptoms in p新鐵iglets，PHEV leads 人人to vomit and neurological sympt大計oms in piglets，爸醫while the other four kin公北ds of viruses cause diarrhea in s信靜wine.Since 2011，a total吃放 of 2 915 samples（intestine做這s or feces）of pi哥上gs with diarrhea 花遠symptom from 15 prov的答inces in China have been test銀道ed for PEDV and TGEV by China朋輛 Animal Health and姐那 Epidemiology Center，and 喝遠1 223 samples were P術家EDV-positive and 109 were T白大GEV-positive. T在呢he proportion of PE報新DV-positive samples 章科and that of TGE亮些V-positive samples of all the個友 sample tested were 費街41.96%，3.74% resp新很ectively. Since 2笑通014，a total of 12 430 t遠文issue（lymph nodes or tonsils）samples of短女 pigs in slaughterhouses from 14開黑 provinces have bee線風n tested for PEDV，and 325森業 samples were PEDV-positive，the 術身proportion of which was 2.61%. Retrosp影員ective detection of 20飛不19-nCoV，which causes novel coronavirus 坐妹pneumonia，was u日風ndertaken. A total 窗地of 2 658 tissue sam短得ples of pigs collected from 15 provinc為西es during 2018 to 2司吃019 were tested to be all 熱理2019-nCoV negative.The高熱 homology between 2019-nCoV生讀 and swine-origin coron業站aviruses was analyz子土ed. The phylogenetic e爸對volutionary analysis based on genome i機下ndicated that the nucleotide homology 做鐵between 2019-nCoV and PHEV both of whi物亮ch belong to Betacoro歌在navirus was only 5公風4%，the other swine-都森origin coronaviruses whi間線ch belong to Alph明制acoronavirus or Deltacoronavirus exhibi兒個ted a longer genetic distance from 自放2019-nCoV. According to the da劇玩ta and analysis above，th務開e possibility of 2019-nCoV origi美熱nating from the known 上亮swine-origin coronaviruses could be 微冷preliminarily ruled out. 全文下載鏈接：ht相時tps://kns.cnki.net/KC子行MS/detail/detail.aspx?dbcode=CJFQ&db刀司name=CJFDAUTO&filename=ZGDW202飛謝003001&v=MTkzMjJyUG兵行ViRzRITkhNckk5RlpZUjhlWDFMd用現XhZUzdEaDFUM3FUcldNMUZyQ1VSN3FmWk靜煙9kdEZ5amtVYnJKUHk=
2020-03-12
基于RPA的鯉春病毒血症病毒核酸檢測技術的建立與評價
重組酶聚合酶擴增技術（recombin拍司ase polymerase amplification，RPA ）是近幾年新用鐵發(fā)展并普及應用的一種(zhǒng)核酸擴增技術，能(n為我éng)夠實現檢測設備小型化，從而滿足現場診斷（po校開int-of-care testing，POCT）嗎爸的需求。本研究以鯉春病毒血症病毒（spring viraemi和黑a of carp virus，SVCV） 爲檢測靶标，建立了SVCV的RP購去A檢測技術，并分析了該方法的靈敏性、特異性和穩定性，妹亮評判了該技術應用于POCT的可能(nén工土g)性。結果表明：基于RPA的檢測方法靈敏度較用看高，檢測下限可達4×103 PFU/mL，但比以T廠拍aq酶爲基礎的RT-PCR技術略低；特異性好(hǎo)，未發(fā)現員裡對(duì)其他水生動物疫病病原，如傳染性造血器官壞死病病毒（IHNV）得歌、病毒性出血性敗血症病毒（VH北林SV）、傳染性胰腺壞死病病毒（IPNV）、傳染性鲑魚貧血病病毒（ISAV）等黑購發(fā)生非特異性反應；穩定性良好(hǎo)，3次重複雜裡性試驗的結果近乎一緻。試驗證明，該方法能雨但(néng)夠替代RT-PCR技術，應用于POCT懂白的試劑和儀器設備研發(fā)。 Es黑從tablishment and Evaluation of a N好數ucleic Acid Det她要ection Technique for Spring Vi鐘我raemia of Carp Viru藍票s Based on RPARecombinase歌動 polymerase amplification（RPA）is a kin從高d of nucleic acid amplification meth木高od developed and widely used in re機黃cent years，which could make rele影子vant detection equipments miniatu愛雨rized to satisfy the requi黃時rements of point-of-care testin又一g（POCT）. In this study，bas子間ed on spring viraemia of 說黃carp virus（SVCV），the RPA met什腦hod for detection 費他of SVCV was esta腦吃blished，and its sen文物sitivity，specifici不照ty and stability were analyzed算謝，then the possibil刀鐘ity of applying th畫到is method to POCT關計 was evaluated. The results show男但ed that the method農新 was with high se微算nsitivity，and its lo裡他wer limit detection was 4×103 PF間們U/mL which was slig土朋htly lower than th妹業at of Taq enzyme房畫 based RT-PCR；its specifici土區ty was good，no any non-specific少見 reaction with the pathogens 紙紙of other aquatic animal was fo樹暗und，such as IHN長服V，VHSV，IPNV and 鐘會ISAV；and the results of three rep熱河eated tests were almost the same due區習 to its good stability. In 白錢conclusion，the RPA method 場舞could be used in research of舊件 relevant instrumen舊新ts and reagents for POCT.全文下載鏈接：ht請地tps://kns.cnki.net/KCMS/detail/劇妹detail.aspx?dbcode=CJ遠樹FQ&dbname=CJFDAUTO&filename=醫線ZGDW202002022&v=MDM0MjZHNEhOSE1雜那yWTlIWm9SOGVYMUx1eFlTN0RoM水日VQzcVRyV00xRnJDVVI3cWZaT2R0RnluaFc3M1BQ空懂eXJQZWI=
2020-03-12
雞傳染性支氣管炎病毒實時(shí)熒光RT-PCR舞業檢測方法的建立與初步應用
爲建立一種(zhǒng)快速準确檢測雞傳染性支氣管炎病毒的方法，根據雞站黃傳染性支氣管炎病毒的核衣殼蛋白基因進(jìn)對請行引物和探針設計，建立了一種(zhǒng)能(néng)夠檢鐵快測雞傳染性支氣管炎病毒的實時(shí)熒光RT-PCR方法，并對(du有歌ì)該方法進(jìn)行特異性和靈敏度檢可數測，同時(shí)與國(guó)家标準檢測方法進(jìn)行對(duì)比暗都。結果顯示，本檢測方法特異性好(hǎo)風弟，不與其他相關病毒發(fā)生交叉反應，靈敏度可達到10-4 ng有他/μL，與國(guó)标檢測方法的符合率達習鐘100%。結果表明，本研究建立的檢測方法可以準麗外确鑒定雞傳染性支氣管炎病毒，具有良好(hǎo)的應用前如男景。 Establishment of a Rea年自l-time RT-PCR Assay for Detect校體ing Avian Infectious Bronchitis Vi鐘哥rus and Its Preliminary ApplicationIn開下 order to establish a rapid an國暗d accurate method for detect他訊ing avian infectious bronchi子術tis virus（AIBV），the 著兵primers and probes were 章可designed according to nu跳紅cleocapsid protein gene樹話s of AIBV，and a real-t個厭ime RT-PCR assay was e文高stablished，then its specific器購ity and sensitiv東腦ity were tested a舊你nd compared wit風為h national standa窗少rd. The results sh鄉日owed that no any我議 cross-reaction with other relev上和ant viruses was 路鐵observed by this assay，and 睡北it was with good specificity，high se相著nsitivity（up to 10-4 ng/做了μL），and the compliance rate b也笑etween the assay and the na她錢tional standard wa市南s 100%. A conclusion was g西日iven that the establis要離hed assay could accurately identi玩作fy AIBV，and it would have a goo電紙d prospect in p自他ractice.全文下載鏈接：http理司s://kns.cnki.net/KCMS/detai笑藍l/detail.aspx?dbcode喝窗=CJFQ&dbname=CJFDAUTO&filename=ZGD了照W202002019&v=MTE2MDd麗車TN0RoMVQzcVRyV00xRnJDVVI3cWZaT2R慢資0RnluaFViL09QeXJQZWJH來睡NEhOSE1yWTlFYllSOGVYMUx1eFk=
2020-03-12
豬瘟病毒抗體納米熒光檢測試紙條的臨床應森雨用
爲探究豬瘟病毒抗體納米熒光檢測試會醫紙條的臨床應用效果，對(duì)181份未免疫豬瘟疫苗、慢議71份已免疫豬瘟疫苗的臨床豬血清，采用本研究試紙條和北京世紀元亨公司的EL跳黑ISA試劑盒分别進(jìn)行抗體匠雪檢測，對(duì)4頭存在母源抗體的不同免疫時(shí)間仔豬血清、不同階為南段的豬血清，采用本研究試紙條進(jìn)行抗體檢測，并門亮對(duì)檢測結果進(jìn)行分析統計。結果顯示，美話本研究試紙條與北京世紀元亨公司的北樹ELISA試劑盒的陰性符合率爲工熱100%，陽性符合率爲95.77%，經(jīng)Kappa檢驗兩(liǎng算生)者一緻性良好(hǎo)；初生仔豬首遠員免後(hòu)，豬瘟抗體滴度有所下降，30 d後(hòu)加強雪紙免疫，抗體滴度逐漸升高，試紙條檢測結果與豬瘟抗體衰減變化過(guò)為著程呈正相關；不同階段豬群的豬瘟抗體陽性率均爲100%，抗體平均值較高，離散度水外較整齊，與規模化養豬場合理免疫程序相吻合冷低。綜合表明，本試紙條的臨床應用效果較好(hǎo)，可用于實時(shí)監時術控豬群的豬瘟抗體變化，适用于基層規模化豬場的豬瘟抗體快速檢測。Clin森技ical Application of Nanofluore得理scence Strip for Det報湖ecting Antibodies against Classica們海l Swine Fever VirusIn order to e刀白valuate clinical effects筆理 of nanofluoresc西雨ence strip for detecting 火為antibodies against classica冷小l swine fever virus（CSFV）懂商，the clinical serum sample訊熱s were collected要錯 from 181 pigs without vaccinati美小on against CSF and 71 pigs w請謝ith vaccination to dete爸和ct corresponding antib現錢ody by the researched 都可nanofluorescence strip歌一s and ELISA kits made森話 by Beijing Anheal Labora有他tories Co.，Ltd，respectiv服音ely，serum samples from 書市four piglets with mater靜海nal antibody at 間票different vaccination ti綠風me and at different stages were detec用黃ted by the strip，then the r鄉時esults were analyzed as follows：th也坐e negative coincidence rate of the厭生 two methods was 100%多好，their positive coin外區cidence rate was 95.77%，and their con煙開sistency was goo暗金d by Kappa identity test不裡；the antibody titer of CSF in n下信ewborn piglets decr吃我eased after their first數鐵 vaccination，an刀刀d gradually increased after 30 days wh藍街en booster vaccin林些ation was implemented，and the results b這費y strip detecti東森on were positiv內上ely correlated with動銀 the attenuation一我 process of antibodies against CSF員窗V；the positive rate of antibody in 空現pig herds at diffe唱木rent stages was 100%，the a相什verage value of antibody tit場亮er was high，and the dispersion was neat車頻，which was consistent with森大 the reasonable immunization proce間市dure in large-scale swine farms. T要年herefore，the changes of antibodies a厭腦gainst CSFV in pig 廠國herds could be monitored in rea煙木l time by this nanofluorescence s跳唱trip due to its good clinical effects，a店用nd it was suitable for媽街 rapid detectio那妹n of CSFV antibody in large-scale f線大arms at the grass-roots. 全文下載鏈接：樂和https://kns.cnki.net/KC子區MS/detail/detail.aspx?dbcode=知農CJFQ&dbname=CJFDAUTO&filename=ZGD玩高W202002018&v=Mz跳我IwNjZZUzdEaDFUM3FUcldNMUZyQ1VS電哥N3FmWk9kdEZ5bmhV務姐cnpJUHlyUGViRzRITkhNclk5RWJ南嗎JUjhlWDFMdXg=
2020-03-12
湖南省4株豬繁殖與呼吸綜合征病毒全基話還因組序列測定與分析
爲了解湖南省豬繁殖與呼吸綜合征病毒（PRRSV）的遺傳變異情況，對(duì)制車湖南省2016—2019年分離的4株PRRSV（Hunan南時/2016/075、Hunan說醫/2017/085、Hunan/雪舞2018/123、Hunan/2019/055）進(j下老ìn)行全基因組測序，對(duì)其ORF5鐵問、NSP2基因以及全基因組核苷酸序列進(jìn)行人開分析。結果顯示，分離的4株病毒株均爲美洲型，全基因組同源性爲83.國北1%~98.7%，其中3株屬于高緻病型，1些有株爲屬于類NADC30型。Hunan/2綠理016/075、Hunan/2017/085店跳毒株全長(cháng)序列與譜系5中亞系5.1的代表性毒株BJ-4親緣放物關系相近，聚爲一簇；Hunan/2018/123毒株與譜系8.7中的tj朋分nh1501毒株聚爲一簇；Hunan/2019/055毒株獨立成(chén樂路g)一簇，與譜系1親緣關系相近。本研究掌握了湖南省PRRSV的吧章遺傳變異情況，從而爲制定有效的防控策略提供了依據。坐微 Determination and Analysis on the Com內雪plete Genomic Sequence of Four Strai體是ns of PRRS Viru相土s in Hunan ProvinceIn order to recogniz司討e the genetic variati放購on of porcine reproductive an個她d respiratory syndrome virus（PRRSV）in 機如Hunan Province，4 strains你短 of PRRSV（Hunan/2016/075，H妹放unan/2017/085，Hu站公nan/2018/123 and Hunan/2019/055）isola讀謝ted from 2016 to 2019 in the provinc為章e were sequenced，and thei熱麗r ORF5 and Nsp2 genes and nucleotide s購科equences of their complete gene were a得議nalyzed. The results showed開購 that all the four strains belonge雨小d into North American（NA）type，with年說 the homology of complete genome of 離作83.1% to 98.7%，in which器購，3 strains were highly patho廠議genic and 1 strain was clas照內sified as NADC30-去輛like strain. In view of the gene訊生tic relationship，the full-length 坐錯sequences of Hunan/2016/075 and Hu人離nan/2017/085 were similar to BJ-吃刀4 strain，the rep體報resentative one of 5.1 substr業少ain of 5 family，which were clustered北計 into one cluste在報r；Hunan/2018/123 was clustered tog熱放ether with tjnh1501 strain 拍兒of 8.7 family；and Hunan/2019/05山鄉5 formed one cluster站吃 by itself，which was similar to 1 f資農amily concerning its gene在學tic relationship. In shor可木t，the status of genetic variation of腦街 PRRSV in Hunan Pr窗村ovince was identi我票fied，which would provide some refere金月nces for developing effe車樂ctive control mea討草sures. 全文下載鏈接：https://kns.cnki.n著媽et/KCMS/detail/detail.aspx?dbcod對習e=CJFQ&dbname=CJ好用FDAUTO&filename=ZGDW202002016&v=MTE1Nz年刀ZWNzNMUHlyUGViRzRITkhNclk5R開年VlvUjhlWDFMdXhZUzdEaDFUM3F從船UcldNMUZyQ1VSN3FmWk數妹9kdEZ5bmc=
2020-03-12
禽源冠狀病毒監測簡報
冠狀病毒（Coronaviruses）在分類地位上屬于套式病毒目（N術區idovirales）冠狀病毒科（Coronaviridae）正冠狀病毒錢月亞科（Orthocoronavirinae），包括α、β、γ和δ等4個屬他工，可感染包括人類在内的多種(zhǒng)計亮動物。其中，α冠狀病毒屬主要感染人和豬、犬、貓、蝙蝠等，β冠狀病毒屬主要感染新金人和牛、馬、豬、鼠、蝙蝠等哺乳動物，γ冠狀病毒屬主用信要感染雞、火雞、鴨、鵝等禽類，δ冠狀病毒屬主要感染野禽和豬。引起(qǐ學歌)此次新型冠狀病毒肺炎疫情的病毒（2019-nCoV）屬市資于冠狀病毒科β冠狀病毒屬。可感染禽類的冠狀病毒主要來自γ冠狀村子病毒屬和δ冠狀病毒屬。雞傳染性支氣管炎病毒（IBV）等家禽中分吃兵離到的冠狀病毒均爲γ冠狀病毒屬。家禽冠狀病毒感染危短男害較大的是IBV。此外，鴨冠狀病毒、鵝冠狀病毒和窗那鴿冠狀病毒等在家禽中也有檢出。一些野禽可感染δ屬冠狀病毒（如鵝口瘡冠狀笑說病毒HKU12等）。2013年著厭以來，北京市延慶區農業局（北京市延她城慶區動物衛生監督管理局）對(duì)全國(guó)17省份共計5 249份們和家禽樣(yàng)品進(jìn)行了禽源冠狀黑下病毒檢測，共檢測到IBV 446株，新型鴨冠狀有山病毒和新型鴿冠狀病毒等新型禽冠狀病毒共277株下報，IBV、新型鴨冠狀病毒、新型鴿冠狀病毒的檢出率西民分别爲8.50%、2.04%、3線什.24%。對(duì)2019-nCoV與禽源冠狀病毒的親緣關系進(j工讀ìn)行分析發(fā)現，201亮文9-nCoV與已知的禽源冠狀病毒核苷劇村酸同源性較低；基于冠狀病毒1ab基因保守區域的進(jìn)化分析表明，2公影019-nCoV與禽源冠狀病毒親緣關系較遠。基于上述分析，初請著步排除2019-nCoV來源于已知湖如家禽冠狀病毒的可能(néng)性。Over什相view on the Surveillance of Avian雜河-orgin Coronaviru志話sesCoronaviruses belong to 如不Nidovirales，family Coronaviridae，Or兵讀thocoronavirinae，which are divided他妹 into four genera兒廠：Alphacoronavirus，Betacoron議又avirus，Gammacoronaviru男水s and Deltacoronaviru紅哥s，and can infect many animals includin司明g humans. Among t歌短hem，Alphacoronaviru照黃s mainly infects 報門humans and pigs，dogs，cats，ba舊西ts，etc.，Betacoronavirus mainly infects著費 humans，cattle，horses懂銀，pigs，mice，bats and other ma吧道mmals，and Gammacoronavirus哥我 mainly infects c拍樂hickens，turkeys，ducks，gee飛說se and other birds，Deltacoronavirus m樂月ainly infects wild birds a黃信nd pigs. The em快我erging 2019-nCoV causing novel coro暗動navirus pneumonia（NCP）out吃男break in China belongs to the Betaco書頻ronavirus of Coronaviridae.坐電 Coronaviruses that can infect 山水birds are mainly from the Gamm匠暗acoronavirus and Deltacoronav章可irus of Coronavir火雪idae. Coronaviruses 靜知isolated from poul木視try such as avian infectious b玩問ronchitis virus（IBV）are all Gammacor話暗onavirus. In the自東 avian coronaviruses，IBV is more 物友harmful to the pou到費ltry. In addition，duck cor舞笑onavirus，goose coronavirus and pi短制geon coronavirus we人慢re also detected. Some wild bi化討rds can be infected with Deltaco物跳ronavirus（such as thrush c很讀oronavirus HKU12，地河etc.）. Since 2013，a total of 5哥火 249 poultry samples fr裡近om 17 provinces in短短 China have been detected b不服y China Animal Health a好如nd Epidemiology Center，a total of 44上匠6 avian infectious bro拍懂nchitis viruses，277 of novel duc問爸k coronaviruses and novel pigeon事個 coronaviruses have been dete算明cted. The positive ra照作tes of avian inf對朋ectious bronchitis virus，no區玩vel duck coronavirus，and知裡 novel pigeon coronavirus were 8音海.50%，2.04% and 3.24%，respec樹腦tively.Analysis of the relationship b樂術etween 2019-nCoV an購器d avian coronavirus found that the n刀媽ucleotide homology between 2019-舊下nCoV and known avian-orgin coronavir麗美uses was quite lower co舞但mpared with mammal-origin coronavi通錯ruses. Evolutionary analysis based on 視兵the conserved region of 村如the coronavirus 1ab ge子議ne shows that 2019-n小子CoV is far from avian 技紅coronavirus. Accord月鐵ing to the above analysis res校城ults，the possibility of 2019-nCoV ori服個ginating from th慢腦e known avian-or市地igin coronaviruses can be preliminar話著ily ruled out.全文下載鏈接：https://吃務kns.cnki.net/KCMS/detail歌懂/37.1246.S.2020老低0206.1649.002.html
2020-03-12
魚類病原性弧菌血清學(xué)診斷芯片技術構建
針對(duì)魚類緻病性腸道(dào)弧亮得菌（Vibrio ichthyoenteri）、鳗弧菌（Vibrio 學煙anguillarum）、副溶血弧菌（Vibrio para現月haemolyticus）、溶藻弧菌（Vibrio alginolyticus知林）、河流弧菌（Vibrio fluvi是少alis）和哈維氏弧菌（Vibrio harveyi），制備了牙鲆（Para舞件lichthys olivaceus）抗血上那清，提取了各弧菌的外膜蛋白、胞外産物、鞭毛蛋白和全菌蛋白，分析了牙鲆抗血清與各很喝抗原蛋白的免疫反應，發(fā)現各弧菌抗血清與各自的抗哥體原反應最強，與其他弧菌抗原有不同程度的交叉反應。將媽來(jiāng)6種(zhǒng)那術弧菌的抗原組分固定于芯片載體形成(chéng)讀筆微陣列，構建了其血清學(xué)診斷抗原芯片，使用辣根過(舞些guò)氧化物酶标記的抗牙鲆IgM單抗2D8作爲檢測抗體，确定了檢務男測結果判讀方法；將(jiāng)抗原芯片應用于牙鲆和大菱鲆（Scophthal老地mus maximus）的弧菌病診斷，并利用ELISA技術，驗證校頻了該抗原芯片用于弧菌病診斷的準确性。本研究爲魚類弧菌用事病的血清學(xué)診斷提供了有力工具。學站Construction of Serological Diagnostic舞多 Chip for Pathogen司身ic Vibrio in FishThe antiserum sampl新家es of Paralichthys olivaceu錯輛s were prepared resp多明ectively against 6 kinds 水愛of pathogenic vibrios，incl秒要uding Vibrio ic暗一hthyoenteri，V. anguillarum，V. paraha分老emolyticus，V. alginolyticus，V. fluvi不請alis and V. harveyi. 都睡The outer membrane pr森快otein，flagellum pr拍從otein，extracellular products and wh弟這ole-cell bacterial protein of the s南技trains were extracted a能時nd purified，then the immune reactions 這歌between the antigen com和姐ponents and the antiserum samples were城我 analyzed，it was found that，for 通民all Vibrios，the immune reaction be做下tween its antigen and舞長 its own antiserum w唱器as the strongest，besides，there were a外低lso cross-reacti件近ons in different 笑明extent between the antiser慢樹um with the antigen 從但of other Vibrios. Further，the a道友ntigen components of the 6 stra書年ins were fixed on the c票行hip carrier to form an an子電tigen microarray so as to construct亮我 the serological diagnostic an員門tigen chip，the method for interpreting中個 detection results was determined by 通在using monoclona多好l antibody 2D8 of IgM marked with舞好 horseradish peroxi林雜dase as the detection 空視antibody；the antigen chip was applied 麗快to diagnose vibriosis in Paralichthys o綠多livaceus and Scophthalmus maximu匠靜s，and its accuracy was verified by文就 ELISA. As a conclusi短頻on，a useful tool was provided for s討得erological diagno黑空sis of fish vib匠民riosis.全文下載鏈接：https://kns.cnki.n科小et/KCMS/detail/37.1246.S.我你20191206.0907.036.html
2019-12-25
羊種(zhǒng)布魯氏菌Rev.1株VjbR基因的克隆、原核表達及生物信息到街學(xué)分析
本研究旨在構建表達載體pET-30a-VjbR，進(jìn)而表達布魯氏菌草男VjbR蛋白；利用生物軟件分析該蛋白生物信息學(xué)信息，爲進(jìn)鄉看一步研究該蛋白奠定基礎。以布魯氏菌Rev.1株基因物離組爲模闆，擴增VjbR基因序列，將內如(jiāng)其克隆至原核表達載體pET-3市問0a（+）中，獲得重組質粒pET-30小的a-VjbR，并進(jìn)行原核表達、Western-blot檢窗但測及該基因的生物信息學(xué)分為窗析。結果顯示：本試驗成(chéng)功克隆并表達了VjbR基通去因；經(jīng)對(duì)表達産物暗黑進(jìn)行SDS-PAGE分析純化，發(fā)現本試驗得到生新較純蛋白；經(jīng)Western-blot檢測，發(fā)現該基因表到書達蛋白可與布魯氏菌羊陽性血清發(fā)生特異性反雪妹應，具有良好(hǎo)的反應原性；通過(guò)生物信息學(xué)系列軟件統藍錢計分析，發(fā)現VjbR蛋白無跨膜區，無信号肽，且該蛋白共有15個抗原決來業定簇，在二級結構中，α-螺旋的氨基酸有131個，占比爲49.81%。影地布魯氏菌VjbR基因的成(chéng)功樂睡表達與純化，爲進(jìn)一步制厭黑備該蛋白的單克隆抗體及iELISA診斷試劑盒的研制奠定了基礎房黃。Cloning，Prokaryotic Expressi睡窗on and Bioinformatic為務s Analysis of VjbR Gene of Brucella 器有melitensis Rev.1 Str如你ainIn order to construct the expressi服制on vector（pET-30a-VjbR）海水to express the VjbR protei訊街n of Brucella，and to analyze the b多雪ioinformatics information of草如 the protein by biologica快現l software so as to lay a 通章foundation for further relevant多短 study. Taking the genome of 歌樹Brucella melitensis員雪 Rev.1 strain a冷花s a template，VjbR gene se她哥quence was amplified a湖森nd then cloned into the p厭件rokaryotic expre個那ssion vector，pET-30a（+），to obtain 見她the recombinant plasmid，pET-30a-Vjb錢兒R，then the prokaryotic expression，wes關銀tern-blot and bioinforma微兵tics analysis were 還不respectively carried out. The results s聽長howed that VjbR gene was su好問ccessfully cloned and expr影姐essed；the purer protein was re音地ceived after SDS-PAGE analysis and 紙議purification for the expr來知essed products；it was d多銀etected that th市件e expressed protein cou要費ld specially react with the p子制ositive sheep serum through western遠兵-blot，showing good reacti熱電vity；according to the analysi路制s by series of bioinf司路ormatics software，it was found 習她that VjbR protein was with no any tran章紅smembrane domain or signal peptide，it h吧件ad 15 antigenic determinants. In the 懂從secondary structure，the number of α-廠窗spiral amino acids was 13煙藍，accounting for 49.81%. In懂雨 conclusion，the successful exp個制ression and purification of VjbR gene少劇 of Brucella would la費著y a foundation 又們for further preparation of mo上醫noclonal antibodies and商在 development of i快制ELISA kit.全文下載鏈接：https:/大務/kns.cnki.net/KC老農MS/detail/37.1246.S.20191206.0907.030.笑山html
2019-12-16
禽腎炎病毒與雞細小病毒雙重PCR檢測方法的建立
爲了同時(shí)檢測禽腎炎病毒（avian nep中頻hritis virus，ANV）和雞細小資機病毒（chicken parvov很信irus，ChPV），根據GenBank中ANV的ORF基因大海和ChPV的 NS1基因的保守序列，設計篩選了擴增玩理片段大小分别爲511 bp和242 bp的特異性引物，通過(guò)紙光反應條件優化、特異性和敏感性試驗，建立了一種(zhǒng事我)雙重PCR檢測方法。該方法最佳舞低引物比例爲ANV上下引物各0.4 μL（20 mol/L），ChPV上的歌下引物各0.6 μL（20 mol/L說科）；最佳退火溫度爲53.8℃；靈敏度可達到55 fg（ANV）裡她和58 fg（ChPV）；對(d銀視uì)常見雞病病原體進(jìn)行檢測，結果全爲陰性。本研究建立遠時的PCR方法具有特異、敏感、快速、穩定的優點，可爸的用于同時(shí)鑒别診斷ANV和ChPV的混合感染。Establish光做ment of a Duplex PCR Assay 就睡for Detection of Avian Nephritis Viru要要s and Chicken ParvovirusA dupl錯朋ex PCR assay for sim山哥ultaneous detection of both a你要vian nephritis virus（ANV）and chicken 窗人parvovirus（ChPV）was establis廠樹hed. Specifically，one pair視要 of specific primers又廠 were designed according 醫師to the conserved sequenc校內es of ORF gene of ANV and NS1 gene o說我f ChPV published in GenBan通學k，after optimization of re資街action conditions，specificity and se讀紅nsitivity tests，the duplex P議開CR assay was establi南長shed and evaluated. The re她費sults showed that the best rati中聽o of primers was 0.間知4 μL（20 mol/L）respe物工ctively for uppe讀校r and lower primers of火也 ANV，and 0.6 μL（20 mol/L）respectively頻近 for those of ChPV；th遠窗e best annealing temperature wa老明s 53.8 ℃；the detection li街筆mits could be up to 55 些近fg（ANV）and 58 fg（ChPV）但到；then the assay was used to det相聽ect common pathogens of chicken diseas鐵空es，and all detect土高ion results were negative. A conclus工做ion was given that，the estab舞頻lished assay was specific，他知sensitive，rapid and stabl習現e，and the assay could be used to iden暗票tify and diagnose mixed infection 好自of ANV and ChPV.店志全文下載鏈接：https://kns公城.cnki.net/KCMS/detail/姐南37.1246.S.20191206.0907.026.html
2019-12-16
羊傳染性膿疱病毒NZ2毒株CBP蛋白制音結構分析與表位預測
本研究應用生物信息學(xué)方法，服費以ORFV NZ2毒株的CBP蛋白氨基酸序列爲研究對(到去duì)象，對(duì)CBP蛋白的理化特性、親水性、跨都地膜區、信号肽、二級結構、三級結構、B細胞表位和T細胞表位進得家(jìn)行了預測分析。結果顯示，CBP蛋白的相對(duì)分子個線質量爲31.179 89 kDa，理論等電點爲4.68，半衰期爲30 h新嗎，穩定系數爲43.06，脂肪族指數爲81.78，總平均親水制熱性爲-0.383。CBP蛋白爲親水性蛋白，無跨膜區，不屬于跨膜蛋白和坐，存在信号肽；二級結構中α螺旋、西樂β折疊、β轉角、無規卷曲分别占38.11%、33.91%、5.94%和10.科事48%，三級結構形似倒立的啞鈴，上下對(duì)稱。篩選出了6個優勢B細胞身又表位、3個CTL細胞表位，無Th細胞表位。CBP蛋好笑白爲親水性膜外蛋白，細胞表位較少，在細胞膜外爲ORFV逃避宿外懂主免疫反應發(fā)揮作用。本研究爲CBP蛋白的功能(né媽自ng)研究與ORFV的緻病機制研究提媽道供了理論依據。Structural Ana看電lysis and Epitope Pre子關diction of CBP of ORFV NZ2 Str票月ainTaking the amino acid身照 sequence of CBP of orf virus（ORFV）朋見NZ2 strain as the research object，t了物he physicochemical property，火明hydrophilicity，transmembrane 土制domain，signal p線大eptide，secondary structure，terti愛拿ary structure and epitop老小es of B cell and 服空T cell，were predicted and analyze家來d by bioinformatics method. T體城he results show坐生ed that the relati放兒ve molecular weight關草 of CBP was 31.179 89 kDa，the theo就都retical isoelectric poin機書t was 4.68，half-life period was 30 問風hours，stability coefficient was 43.0從票6，aliphatic index was 81.78，a愛個nd total average hydrophobicity was 火近-0.383. CBP was 有答a hydrophilic protein instead 城山of a transmembrane one，no any輛玩 transmembrane domain，but signa道山l peptide was con木物tained；α-helix，β-fold，β-turn可草 and random coil accoun黑友ted for 38.11%，33.91%道區，5.94% and 10.48% respectively in 很什the secondary structure；the錢坐 tertiary structure was simil外議ar with an inverted議件 dumbbell in for厭會m，and it was symmetrical f飛議rom top to botto雪時m. A total of 6 dominant 工區B cell epitopes and 知上3 CTL cell epitopes were screened o民不ut，and no Th cell epitope wa高線s found. CBP was a hydrophi對線lic extracellul在音ar protein with few c雜看ell epitopes，which he但生lped ORFV avoid host immune r男農esponse outside the c刀村ell membrane. In conclusion，a theoretic些很al basis was pro少唱vided for the study on CBP' s func通身tions and the pathogenesis of 國國ORFV.全文下載鏈接：https:/近數/kns.cnki.net/KCMS/detail/輛說37.1246.S.20191206.0907.032錢河.html
2019-12-16
非洲豬瘟病毒結構蛋白p54的原核表喝也達與反應原性鑒定
p54蛋白爲非洲豬瘟病毒（ASF近輛V）的主要結構蛋白之一，參與病毒對(duì)靶細胞的吸附與進(jìn)入。爲亮空深入研究p54結構蛋白的抗原性，輛空根據GenBank序列号（MK128995）對(duì錢女)應的E183L基因序列，設計1對(duì章服)特異性引物擴增其整個CDS區，并克隆至pET-30a（+）載體，靜睡構建了表達p54蛋白的重組質粒pET-30a（+）-p54。用BL21（又文DE3）轉化該質粒，經(jīng影船)IPTG誘導表達後(hòu)進(j工你ìn)行SDS-PAGE電泳，可見重組報生質粒表達出1條分子量約爲20 kDa的特異性條帶，且重組表達蛋司務白以融合表達蛋白形式存在于上清。但就進(jìn)一步通過(guò)His親和層析法純化目的蛋白，用ASFV火校陽性血清進(jìn)行蛋白質免疫印迹音遠反應，發(fā)現表達的重組p54蛋白能(néng)與ASFV陽性兵議血清産生特異性反應，表明p54蛋白表達成(chéng)功。本研究爲ASF下市V抗體ELISA檢測方法的建立奠定了基礎。Prokaryotic Expre文信ssion and Reactivity Identificati地紙onof Structural Protein p54 of作美 African Swine Fever VirusAs o歌工ne of the major st店冷ructural proteins of對用 African swine fever校學 virus（ASFV），p54 pr從唱otein helps the vi畫北rus adsorb and ente司要r into the target cells. In order t土亮o further study i我湖ts antigenicity，one道器 pair of specific p拍到rimers were desig校大ned to amplify the whole 坐微CDS domain of E183L gene ac男議cording to the 金資serial number（MK128公樹995）in GenBank. Then劇信 the amplificati女火on products were cloned 兒民into the vector pET-中科30a（+），and the recombinant plasmid p制車ET-30a（+）-p54 was constructed，af鐘喝ter transformation in這很to BL21（DE3），in畫黃duced expression by IPTG and SDS匠兒-PAGE electroph個劇oresis. one specific band wi村你th the molecular weight 兵微of 20 kDa was obtaine影暗d and the recombinan很師t protein appeared in th小筆e supernatant in the form of fu金拍sion protein. Furth樹科er，the recombinant pr錯花otein was purified by His-tag 笑村affinity chromatography，an市哥d the positive serum of ASFV was use市志d for Western blotting. it was found 技影that the expressed內站 recombinant p54 protein could 區答specifically react with ASFV posi站數tive serum，indicating th近民at p54 protein was successfully e章作xpressed. The study woul對朋d lay a foundat文做ion for the development of ELISA 工公kit for detection of AS個國FV.全文下載鏈接：https://kn樹生s.cnki.net/KCMS/detail/秒們37.1246.S.20191206.0907.028.html
2019-12-16
2018年遼甯省大連市奶牛副結核病血清什明流行率調查
本研究對(duì)大連市全部11個奶牛場開(kāi)展抽樣(yà美讀ng)調查，通過(guò)設計兩(liǎng)階我路段随機抽樣(yàng)方案并結合問卷調查，初步了解喝鐵了大連市奶牛場副結核病流行狀況以及奶牛場從業人員對(duì)副結核病男門防控知識的掌握情況和行爲特點。研究結果顯示，大連市奶牛場副弟區結核病平均個體血清流行率爲17.2森會3%（95%CI：15.99%~18.74%數靜），場群真實流行率爲100%；年齡、路河胎次是奶牛副結核病重要的風險因素；奶牛副結核病是導緻大連市奶牛腹瀉的主要原因之中議一；大連市奶牛場從業人員對(duì)副結核病的認知程度有限，部分規模場對(du低離ì)本病認識不足，導緻部分防控措施落實不到位。研究結果提示，大連市需重做水視對(duì)奶牛副結核病的防控，采取綜合防控措施，阻止他很本病的繼續蔓延并在有條件的場戶開(k師嗎āi)展淨化；各奶牛場應嚴格做好(hǎo車志)生物安全工作，在引進(jìn)奶牛時(shí)嚴格去都檢疫，防止引進(jìn)陽性牛、隐性感染牛；獸醫部門應該加強科普宣傳，增黑靜強奶牛場工作人員對(duì)副結核河員病的判斷力，提高其防範副結核病的能(néng)力。I在信nvestigation on Sero-林白prevalence of Cow區鄉 Para-tuberculosisin Dali熱電an City of Liaoning Province in 車男2018In this study，a這麗ll the 11 dairy farms in Dalian 不中City were sampled 站商and investigated，and the prev技議alence of para-tuberculosis，practitio影討ners' knowledge and beh刀業aviors for control of the d時從isease were prelim為還inarily identified through two-st知車age random sampling pla分妹n and questionnaire光內 survey. The results sh金間owed that the av森公erage individual說做 serological prevalence of para-tu家弟berculosis was 17.23%（95%CI：15.99%–1下他8.74%），and the true prevalence at the船秒 farm/herd level was 100%；the ris習冷k factors of such infection i紙服ncluded age and calving number；cow 唱錯para-tuberculosis多微 was one of the main causes of diarr請村hea；and some control measures c美你ould not be put in place due t人拿o lack of relevant knowl請志edge and awareness in some scaled far師了ms. Therefore，it was suggested that t到兒he disease should be controlle去的d as a top prio朋媽rity by series of comprehensive measu哥快res to prevent any further spreading玩拍，some purification measures could be兵南 carried out in some conditional 村我farms/households；bio-safety measures s小機hould be carried out by all farms，al通門l introduced co時高ws should be strictly quaran近家tined to avoid any p窗著ositive cases or 年人suppressive infection；al懂白so，popularization and pu國和blicity should be strengthened by ve海業terinary departm明年ents to enhance workers'新拍 judgment on the diseas船火e in the farms and歌業 to improve their abili暗月ty to prevent the disease.全文下載鏈接：事也https://kns.cnki.net/KCMS/detail/37.12匠術46.S.20191206.0906.008.h公明tmll
2019-12-16
H9亞型禽流感病毒實時(shí)玩離熒光RT-PCR檢測方法的建立與應用
爲建立一種(zhǒng)快速準确檢測H9亞型禽流感病毒基因的檢測方法，根據G白妹enBank中H9亞型禽流感病毒血凝素編碼基因進筆懂(jìn)行熒光檢測引物和探針設計，建立了一種白你(zhǒng)針對(duì)H9亞型禽流的愛感病毒的熒光RT-PCR檢測方法。利用該方法進車間(jìn)行靈敏度和特異性檢測，并用本方法鐘東和國(guó)家标準中的熒光RT-PCR檢測方法，同時(shí)對(duì就如)臨床樣(yàng)本進(jìn)行檢測。結果顯示，僅H9船員亞型禽流感病毒出現了正常熒光檢測曲線，而其他病毒及陰性對(duì)照均近黑未出現；檢測靈敏度爲RNA終濃度10-4 ng/μL（1.30×104 co呢多pies/μL）；臨床樣(yàng)本檢測結果與長視國(guó)标方法一緻，符合率爲100%。笑分結果表明，該方法特異性強、靈敏度高，可用于H9亞型禽流感病毒報子檢測。Establishment and人生 Application of a Real-time RT-話個PCR Assay for Detection志多of H9 Subtype Avian Influenza Virus分說In order to establis生唱h a method for 事著detecting H9 subtype avia離睡n influenza virus（身明AIV-H9）rapidly and accurately，短費the fluorescence detection prim友好ers and probes were designed and syn街著thesized according to hemagglutinin見下 encoding gene 錯習of AIV-H9 registered i年快n GenBank，and a fluorescence PCR assa音間y was thereby established，a新會nd its specificity and se鄉短nsitivity were tested. Then by th新森e established assay and the RT-P有中CR stipulated in national standard，s紙電ome clinical samples were simultaneousl會船y tested. The res兵要ults showed that the 師話normal fluorescence detection cur坐頻ve only appeared in AIV-H9 rather t音影han other non-specific viruses a西文nd negative sample；its detection li音街mit was 10-4 ng/μ舞刀L（1.30×104 copies/μL）of RNA；the detec用呢tion result of clinical samples一麗 by established ass雪弟ay was consistent with t術光he national standard，and the co飛煙incidence rate was 100%. In 我飛conclusion，the assay could be used to d計小etect AIV-H9 due to its s船著trong specificity and 白的high sensitivity.全文下載鏈接：https://kn中村s.cnki.net/KCMS/deta懂農il/37.1246.S.20行懂191206.0907.024.html
2019-12-16
2018年海南省豬A型塞尼卡病毒血清流行業慢病學(xué)調查
爲了解海南省A型塞尼卡病毒（Senecavirus A，S你個VA）感染狀況，應用間接ELISA方法，對(d志人uì)從海南省15個市（縣）112他地個場點采集的2 547份豬血清樣(yàng)品進(jìn)行SVA抗體檢音輛測。結果顯示：SVA血清樣(yàng)品抗體陽性率爲10.9%，明謝場點陽性率爲50.0%；15個市（縣）的場點陽性率生煙在14.3%~100%之間，其中有6個超過(guò)50%；15人醫個市（縣）的樣(yàng)品陽性率在1.0%~31.7%之間，大部分市（縣）厭南小于10%；規模化養殖場場點陽性率（61.2近靜%）高于散養戶（33.3%）；仔豬、母豬、後懂就(hòu)備豬、公豬、育肥豬樣(yàng)品陽性率分别是2動要0.5%、15.5%、13.5%、10.7%、5多年.9%。結果表明：SVA已擴散至整個海南省吃城，防控形勢較嚴峻，需制定相應防控措施；交通樞紐地區和技紅規模化養殖場的SVA流行面(m時很iàn)較廣，仔豬和種(zhǒng)豬群中的SVA流行率較高，應重點加強監樹件測和相關流行病學(xué)研究。本研究摸清了海南省豬群中的SV我舞A的感染與分布情況，找出了重點防女音控區域和防控對(duì)象，對(duì)于指導該地區SVA防控具可司有參考意義。A Seroepidemiolo紙森gical Investigation on Senecavirus高拍 A in Swine in Hainan為土 Province in 2018In order樹家 to identify the prevalence of Se煙器necavirus A（SVA）in H下很ainan Province，2 547 pig 村歌serum samples were collected from如他 112 farms in 15 cities（counties），and S劇可VA antibodies were detec秒森ted by indirect姐現 ELISA. The result筆道s showed that the average po會得sitive rates of SVA s歌有erum antibody at the indiv那愛idual and farm level were 10.9% and 50.場月0% respectively；the positive rates 懂務at the farm level in 15 討會cities（counties）ranged from 媽靜14.3% to 100%，and高雪 the rates in 6 out of 15 cities（co懂好unties）exceeded 50%；the positive rat上計es at the individual level理些 in 15 cities（counties）range家門d from 1.0% to 31.7%，and the rates i醫醫n most samples were less than 10他答%；the positive rate in白技 scaled farms（61.2%）was high了街er than that in fre開視e-range households（33.3%）；th看請e positive rates of s拍但amples from piglets，sows，replacem頻船ent gilts，breeding pigs裡友 and fattening pigs we慢身re 20.5%，15.5%，13.5%，10.7% and 5.9%，r為新espectively. The results indicat林學ed that relevant control meas都務ures should be developed under su工費ch severe situation that SV他外A had spread acro花國ss the province；monitor吃場ing and corresponding epidemiologi討習cal research should be strengt影火hened as the top pri說黑orities due to wide spread of SVA i暗拍n transportation hub a劇睡reas and large-scale farms女站 and its high preva廠公lence in piglets an們間d breeding popula務議tions. In this study，the infect玩東ion and distribution of白來 SVA in pig populations i一不n Hainan Province were identified，and 遠器the key areas and objects 時妹to be controlled were defin窗歌ed，which could provide s謝技ome references for prevention and cont可微rol of SVA in the province.全文下載鏈要白接：https://kns.cnki.理工net/KCMS/detail/37.1246.S.20慢地191206.0906.004.html
2019-12-16
安徽省規模禽場高緻病性禽流感血清弟女學(xué)橫斷面(miàn)動電研究與風險因素分析
爲掌握安徽省禽群的禽流感免疫抗體水平以及引起(q們時ǐ)免疫不合格的主要風險因素，在2018年春季集中免疫前，對畫樂(duì)安徽省7個市開(kāi)展了H5機樂和H7亞型禽流感血清學(xué)檢測和問卷調查。按照分層随機抽樣(yàng很知)方法，在7個市選擇種(zhǒng)禽場、蛋禽場和肉化姐禽場，共140個養禽場，在每個養殖場内随機抽取35份血清樣件對(yàng)品，進(jìn)行HI抗體檢測，并根據檢測結果，文理計算群體水平的真實免疫合格率/陽性率；日輛將(jiāng)H5或H7亞型禽流感抗體群體合格率低于7答房0%的養禽場定義爲免疫不合格場，進(jìn)行單因素就內風險分析。結果顯示：安徽省7個市H5（Re-8株）和H7N9（Re-秒說1株）的場群真實抗體合格率/陽性率分别了雨爲67.7%（95%CI：60.0%~75.4%）和39.8%（95%CI知票：31.7%~47.9%），62個養禽場的H5或從的H7亞型抗體未達标（＜70%）；存欄量見風越小（P=0.046）、免疫次數越少（P=0.016），H5或H7亞型用多免疫抗體不合格的可能(néng)性越高（P＜0.外歌05）；存欄量小于4 000羽、免疫後物空(hòu)時(shí)間≤21 d或≥91 d、家禽日齡≤26或≥2 200新用、免疫次數≤1、規模化飼養鵝，以及不使用“H5+H7”二價滅活疫苗（O月高R=2.66，95%CI：1.34~5.35）、周邊3 km内有野禽公著栖息地（OR=2.78，95%CI友線：1.10~7.65）的養禽場的H5或H7亞型免疫抗體不合格風險相為議對(duì)較高。結果表明：2018年春季安徽省7個市H黃可5、H7亞型禽流感免疫抗體保護率下降幅度明顯，需西數要及時(shí)開(kāi)展春季集中免疫；養禽場規模、類型以及暗和家禽種(zhǒng)類、是否免疫疫苗、免疫次數、養禽場周邊是否有野紅高禽栖息地等，均與群體的禽流感免疫抗體合格率顯著相關，因此在制定強制免疫計劃時(站時shí)，應視情況進(jìn)行分類指導，實時(shí)補免或增來聽加免疫次數。此外，養禽場選址時(shí)應盡量避開(kāi)野禽玩如栖息地。本研究爲決策部門制定禽流感免疫方案提供了技術支撐。A Serologi遠兵cal Cross-sectional Study on HP看要AI in Large-scale Poultry Farmsin A站輛nhui Province and Analysis on雨懂 Relevant Risk FactorsIn order t短裡o identify the leve歌做l of antibody against西些 highly pathogenic avian這市 influenza（HPAI）in large-scale poultry 公黃farms in Anhui Province and to a錢明nalyze major risks that might lead to 資爸unqualified immunity，serological 裡空surveillance and questionnaire 喝快survey on H5 and H7店村 subtype HPAI were 訊劇carried out in 7 cities prior to inten跳務sive vaccination in the spring of 民飛2018. A total of 140 p錢制oultry farms includi空中ng breeding，layer and broiler 金都farms were selected to ran醫近domly collect 35匠站 serum samples per farm using strati北微fied random sampling method for hemaggl妹紙utination inhibition tes員師t（HI），and then the tr校刀ue immune qualification rate/posi這南tive rate at the farm leve但村l was calculated according to th金老e test results；the farms of which the 民兵qualification rate of antibod志員y against H5 or H7 subtype virus w和很as lower than 70% wer議拿e defined as the ones with u在司nqualified immunity，and then risk analy多吧sis on single factor was carried o得樹ut. The results showed that the唱人 true immune quali低樂fication rates/positive 報物rates of H5（Re-8 strain）and H7N9（市作Re-1 strain）at the farm le錯你vel were 67.7%（95%CI：6少腦0.0%-75.4%）and 39.8%（95%CI：31.7%-47.9%就事），respectively；in 62 farms，the愛章 antibody level failed to信白 reach the standard拿站（less than 70%）；the smaller the stock（從光P=0.046）and the less 通麗the vaccination frequency（P=票學0.016），the higher the proba森國bility of disqualification 綠關of immune antibodies（P＜0.05在雪）. The high risk factors were as fol間朋lows：the poultry stock was 放問less than 4 000，the period was within 2中唱1 days or above 91 days黑會 since last vaccination，the ag笑腦es of live poultry 高頻were less than 26 day下就s or more than 2 200 days，the immuniz喝車ation times were less 拍務than or equal to 1，raising ge業女ese in a large scale，t錯又he poultry were not vaccinated with說房“H5+H7”bivalent inactivated va錢又ccines（OR=2.66，95% CI：1.34-土我5.35），as well as there were wi線畫ld bird habitats within 3 道鄉km radius around（OR=2樹好.78，95%CI：1.10-7.65）. R務懂esults showed that，the level場船 of antibody against H5 and些生 H7 subtype HPA上到I decreased obviously in th志放e above 7 cities in the sprin哥弟g of 2018，hence it was necessar嗎師y to carry out intensiv和唱e vaccination in spring 通美in time. The antibod讀輛y level was obv說分iously related to the factors in筆家cluding the scale and跳師 type of poultry farms，the type of po銀舞ultry，vaccination or not，frequenc男得y of vaccination，wild b音學ird habitats around farms，etc.，so rel低又ated vaccination should科服 be strengthened or supp暗拿lemented in real time with classifica一間tion guidance as require土是d when the compulsory immunization 暗長plan was develope哥站d. In addition，any是木 wild bird habitat should b光熱e avoided to the g快我reatest extent w志厭hen a poultry farm was d術兵esigned and constructed. A機話bove all，some t睡北echnical supports f書厭or developing AIV immunization p拿都lan were provided to decision-making 村拿departments.全文下載鏈接：https://kns.cnki微樹.net/KCMS/detail/好美37.1246.S.20191206.0906.002.ht多動ml
2019-12-16
馬梨形蟲病病原雙重熒光PCR快速檢測方法的建立與我都應用
在序列比對(duì)分析的基礎上，設計2對(duì)引制吃物和2條MGB探針，經(jīng)體系優化建立了市她可快速鑒别檢測兩(liǎng)種(zhǒng)在你馬梨形蟲病病原（馬泰勒蟲和驽巴貝斯兵坐蟲）的雙重熒光PCR檢測方法。該方法可分别檢出分玩22和31基因拷貝的蟲體DNA，且不與其他馬病病原發(fā物機)生交叉反應。應用建立的熒光PCR檢測方法，對(duì)采自進(j舞工ìn)出口馬匹的10份馬全血和20份馬血清進(jìn)行檢測，結果從馬微老全血中檢出2份馬泰勒蟲陽性樣(yàng)品。使用一懂懂對(duì)馬泰勒蟲EMA1全基因擴增引物，從2份陽性樣(yàng)品中開長擴增了EMA1基因。克隆測序後(hòu)，對(duì)該基船那因全序列進(jìn)行分析，證實信年其爲馬泰勒蟲特異基因序列；2份樣(yàng)品的基因序列相似性爲98.9%們輛，存在6個氨基酸變異；2個樣(yàng年些)品中的EMA1基因不完全一緻，表明2個感染樣(yàng)品中的蟲體可能愛個(néng)爲不同的馬泰勒蟲蟲株。Establishme問資ntand Application of a Duplex飛資 Real-time PCR Method for Detect拿空ionof Piroplasmosis Di黃購sease Two pairs ofprime城木rs and two MGB probes were明拍 designed based on sequen秒中ce alignment，after system o跳就ptimization，a duplex real-time PCR 姐花method foridentifi輛公cation and detection of two p廠玩athogens（Theileria equiand Babesia 村來caballi）of piroplasmosisdisease was外費 established. Th地器e detection limits were 22 and 31 gen和呢e copiesrespectively，and no cros離離s reaction with 好草otherpathogens of horse diseases 你南was observed. By the established 作不real-time PCR，10 whole b工雪lood and 20 serum samp裡車lescollected from imported and expo很好rted horses were tested，and妹子 2 positive samples（against Theileria可務 equi）were detected；then EMA1 市就gene from the two positivesamp看林les were amplified用在 through a pair of站分 amplification primers of EMA1 gene動南 ofTheileria equi. After being clo長大ned and sequenced，the wholeseq頻男uence of the genes were analyze黑他d，and theamplified genes we這作re confirmed to be the specific s些知equences of Theileria equi；T厭醫he similarity of EMA1 sequence of雨有 thetwo samples was 98.9%，with six a也商minoacid mutations，indica務件ting that the wor學木ms in the twosampl自低es might belong to di湖年fferent strains o微信f Theileria equi.全文下載鏈接：https://k暗開ns.cnki.net/KCMS/detail友麗/37.1246.S.20191009.1108.038請從.html
2019-10-24
病死豬圓環病毒2型檢測及全基因序列分析
爲準确檢測豬圓環病毒2型（PCV2）并分析其基因序列情況，在序員海列比對(duì)分析的基礎上，設計一對(duì)船村簡并引物和一條探針，經(jīng)體系優化建立了PCV物相2的熒光PCR方法。該方法可檢出10拷貝的PCV2質粒DNA，不與豬瘟亮亮病毒等多種(zhǒng)病毒發(fā)生交叉反應。對(duì)送檢的2市志批次45份病死豬髒器進(jìn)行檢能能測，檢出25份陽性樣(yàng)品，與套式PCR方法的複合頻道率爲95.6%；對(duì)3份強陽性樣(yàng)品使用一對(duì)全基白有因組擴增引物，擴增了PCV2型全基因片段；經(jīng妹制)克隆測序後(hòu)分析，發(fā)現3個病毒基因分别屬于2種(zh電費ǒng)基因亞型（PCV2b和PCV2d）。本研究對(duì)于P這樂CV2的準确檢測及其變異特征的了解具有參考意義。Detec年村tion ofPorcine Cir的歌covirus Type 2 in Dead Pigs 來中and Analysis on ItsComplete Genome Sequ村器ence In order to門海accurately detect porcine ci日廠rcovirus type 2（PCV2）and t票也o analyze its g化用enome sequence，a pair of degene刀日rate primers and aprobe were designed火時 based on sequence align月木ment analysis，after systematic optimiza西刀tion，a real-time P自老CR method was established，which 紙短could detect 10 copies ofplasmid DNA爸長 of PCV2，with no anycross react去坐ion with classical swi討農ne fever virus and other viruses. Th鐵科en twobatches of 45 organ 綠好samples collected from dead pigs we內報re tested by established匠大method，and 25 samples were dete離多cted to bepositive，the coincidence rat月跳e with nested-PCRwas 95.6%；for the t如山hree strong-positive sa購山mples，their complete g信紅ene fragments of北腦 PCV2were amplified by a 藍山pair of primers，then weresequenced an嗎服d analyzed，and it wasfound that the g美技enes of the three 司媽viruses were classified into t她少wo differentsubtypes（PCV2b and PCV2d）.些人 Therefore，it was of和數 certain significance toconfirm 錢刀relevant test re拿秒sults and recognize 業生any variation o還拿f PCV2 by the abovemethod. 報照全文下載鏈接：https://kns.cnki.net關志/KCMS/detail/37知林.1246.S.20191009.1107.036.html話月
2019-10-24
PCV2感染後(hòu)JAK-STAT信号通路的基因差異表達分析你自
JAK-STAT信号通路是闡明細胞因子生物學(x吧算ué)功能(néng)及病毒如何發(fā)揮作用的分子基礎，但對(du務要ì)于豬圓環病毒2型（PCV2）感染及病毒複制的作用機制，目前尚學拿無報道(dào)。本研究采用PCR列陣方法，建立PK-15細胞JAK-制聽STAT信号通路的功能(néng)分類芯片，分析JAK-STA商他T信号通路中所有已知的Jak和Sta亮志t家族成(chéng)員、激活JA睡睡K-STAT信号通路的受體，以及與Stat蛋白相關區生的核輔助因子及共同活化因子、Stat誘機工導蛋白及該通路的負反饋調節蛋白等基紅費因在PCV2感染後(hòu)的差異表達情海紙況，爲進(jìn)一步明确PK-15細胞購少JAK-STAT信号通路對(duì)病毒複制力和複制機制奠定基礎城為。Analysis onDifferentia鐵到l Gene Expressi船做on of JAK-STATSignaling Pathway a商雪fter PCV2 InfectionThe離妹 JAK-STATsignaling pathway could he開也lp us to expound biological func場麗tion of cytokines andhow viruses work，科路but the infection with p樂票orcinecircovirus type 2（PCV2）and看聽 virus replication m拍看echanism havenot been reported so far.去看 In this study，PCR array wasused t關中o establish a functional 如男taxonomic chip of the JA拍討K-STAT pathway in PK-15cells，t哥要hen the differential g如議ene expressionof the pathway after PC們房V2 infection were analyzed，includin關說g allknown Jak and Stat family m白會embers，receptorsactivating數他 the pathway，nuclearcofactors an秒城d coactivators related to St少訊at protein，Stat-inducible protein and鐘懂 negativeregulato為綠ry proteins，whic理學h wouldlay a foundati還服on for further identif森下ying how the pathw樹吃ay influence virusreplication and 有金replication mechanism.章歌全文下載鏈接：https://kns.cnki報站.net/KCMS/detail/37街城.1246.S.20191009.1107.0章人34.html
2019-10-24
表達羊口瘡病毒B2L蛋白重組小反刍獸疫病毒的拯救外高
羊口瘡（傳染性膿疱）和小反刍獸疫的病原體分土和别爲羊口瘡病毒（orf virus，O要年RFV）和小反刍獸疫病毒（peste d刀煙espetits ruminants virus，PPRV）。如果通過(gu遠輛ò)反向(xiàng)遺傳技術進(jì坐好n)行基因修飾，PPRV可成(chéng)民藍爲表達外源蛋白的有效載體。ORFV含有1個具有高免疫原性的B2L蛋白。本研究首章購先構建了含B2L基因開(kāi)放閱讀框的重能道組PPRV cDNA clone，然後(hòu)通過(guò)反向(xià物道ng)遺傳技術，拯救了一株表達B2L蛋白的重組PPRV。試驗表明上他，該重組病毒的生長(cháng都地)動力學(xué)類似其母源病毒；Western blo家門t和質譜分析表明，該重組PPRV可在細胞内表達B女厭2L蛋白。該研究爲羊口瘡和小反刍獸疫二聯疫苗的研制奠定了基礎亮頻。Rescue of Recombinant Peste des Pe也員titsRuminants Vir還事us for Expressing Orf V一腦irus B2L ProteinOrf（contagiousecth人相yma）and peste d北風es petits ruminants（PPR）are caused b聽村yorf virus（ORFV）and peste des嗎民petits ruminants virus（PPRV），respect答舞ively. If genetically modifiedb靜水y reverse genetics t答著ool，the PPRV wouldbe a use市明ful vector to express f農水oreign proteins. The ORFV contains飛內 a highlyimmunogenic envelop師問e protein，B2L protein.In this study務頻，a recombinant PPRV cDNA clone，w秒話hich contained the B2L gene open們學reading frame，was re中資scued using reverse genetics toexpress北物 the B2L protein. This recombinant P區影PRV was demonst員西rated to have similargrow術著th kinetics to that of its paren火秒tal virus. In vitro expres知你sion of the B2Lprotein by 制學it was demonstrated b資家y Western blot and confirmed by mas暗議sspectrometry，therefore paving民商 the way fordevelopment of a bivalent鄉睡 vaccine candidate against 如南both diseases.全文下唱商載鏈接：https://kns.cnki.ne民外t/KCMS/detail/3森到7.1246.S.20191009.1107.0吧放32.html
2019-10-24
以GIS爲基礎的禽流感預測預警研究進(jìn)展
GIS是一種(zhǒng)對(duì)地理環境等方面(miàn)高暗數據進(jìn)行儲存、分析、加工和呈現的綜合工具。利用GIS可以把流感件場病例的相關數據與氣候、環境和社會(huì)等數據進(jì知錢n)行整合和分析，通過(guò)統計學(xué)或數學(xué)模型，找到影響如了流感發(fā)生和傳播的風險因素，對(duì)流感的發(fā)生趨勢進媽醫(jìn)行預測和可視化預警。國(guó)内外學(xué)者利用G線鐵IS在禽流感風險因素分析和預測少有等方面(miàn)做了大量研究，月美包括對(duì)人感染H7N9流感、高緻病性H5亞型禽流感、野禽流感風險因務開素和預測的研究，不同基因型病毒分布特征的研究，獸醫決策輔助應用研究等去作。這(zhè)些研究對(duì)指導獸醫流行錢外病學(xué)研究人員將(jiāng)GIS更好(hǎo)地應用于禽流感防控新人具有幫助意義。ResearchProgress劇和 of Forecasting and Early Wa嗎到rning of AvianInfluenza Based on GISGeo師有graphicinformation system（GIS）is些東 a comprehensive tool for storing，匠購analyzing，processing and pre雪懂senting data relatedto g劇畫eographic environment，whic草請h couldintegrate and an公就alyze the data regarding 技北influenza cases，climat答市e，society andothers，and identify森了 all risk factors foroccurre男讀nce and spreading of用花 influenza by use of stat鄉知istics or mathematicalmodels to fo自還recast and visually 公玩warn any trends of influe亮路nza. In recent years，a large num店見ber of studies on risk線國 factorsanalyzing and媽機 forecasting of av身通ian influenza（AI）had been c紙微arried out by schol白間ars inboth home煙們 and abroad by means of GIS，including，s志好tudies on the risk facto輛光rs andforecasting of human infection謝但 with H7N9 influenza，H5 highly p到是athogenic avian influenza（HPAI）and 自慢wild AI，studies on the distributionch舞通aracteristics of 車裡different genot體多ypes of viruses，as well as theappl玩筆ication researches on the s知高upporting of veterinary decis日日ion-making. The化放sestudies would provide some reference船姐s for veterinary epidemiologists toeffe商事ctively control AI by GIS.全文下載鏈接：h區腦ttps://kns.cnki.net/KCMS/detail/37.124機鐘6.S.20191009.1107.028.html
2019-10-24
新疆某豬場母豬繁殖障礙病病原學(xué)檢測和毒株鑒定
2018年4月，新疆某規模化繁育豬場出現大量母豬窗線流産，産死胎、木乃伊胎等繁育障礙症狀。爲确診病因，采集胎兒組織和母豬流産分泌不好物，通過(guò)普通PCR、RT-PCR、real-time PCR技術空跳，對(duì)豬繁殖與呼吸綜合征病毒（PRRSV）、僞狂犬病病毒（P身一RV）、豬細小病毒（PPV）、弓形用南蟲4種(zhǒng)病原進(jìn)行檢測。結果顯示，PRRSV檢測呈陽性，公她PRV、PPV和弓形蟲檢測呈陰性。通過(guò)PRRSV特異性暗他引物NSP2進(jìn)行PRRSV毒株鑒定，确定該豬場存在月林PRRSV類NADC30毒株。結果表明，我弟類NADC30毒株是導緻該豬場出現母豬繁殖障礙短離的重要病原，說(shuō)明該新型毒株已經(jīng)蔓延到新疆地區，并開(關公kāi)始對(duì)該地生豬養相呢殖業造成(chéng)危害。本試驗爲新遠海疆地區豬場母豬繁殖障礙的病因調查做睡提供了依據。Pathogen Detection相作 of Sow Reproductive技算 Disorder Disease and Identification of爸錯 Relevant Strain in a Swine Farm 自通in Xinjiang In April 2018光讀，some symptoms of reproductive 紅湖disorders，such as abortion，stillbirt裡關h and mummified fetus，appeared i懂可n a large number of sows知答 in a scaled br喝計eeding farm in Xinjiang. In 近爸order to identify all related causes，so玩地me fetal tissues and abortiv做亮e secretions from sows were 視動sampled to detect porcine repro線海ductive and respiratory syn開頻drome virus（PRRSV），pseudorabie慢時s virus（PRV），po笑錢rcine parvovirus（PPV）and tox金是oplasma gondii through n懂就ormal PCR，RT-PCR and real吧我 time PCR technology信生 respectively. The res朋那ults showed that the samples were posi影湖tive for PRRSV，and睡習 negative for PRV，PPV and Toxoplasma 知近gondii. The NADC30-like strain of PR店區RSV was identified in the 什鄉farm by NSP2，the speci門拍fic primer of PRRSV. Besides，錢看the reproductive disorders i也要n the farm may be mainly caused by the厭時 NADC30-like strain，indicating錢中 that the new strain had線信 been spreading to Xinjiang and begun 家去to bring great harm to local pi弟弟g-production industr舊藍y. In view of that，some referenc嗎暗es were hereby provi吃可ded for investi答姐gating the causes of sow reproductive 紅子disorders in Xinjiang. 全文下載鏈接：http:/區北/kns.cnki.net/KCMS/detail/37.西學1246.S.20190903.1708.038.html
2019-09-17
2013—2017年我國(guó)小反刍獸疫病毒H基因分子演空呢化特征
爲研究我國(guó)小反刍獸疫病毒（PPR妹會V）的分子流行特點，對(duì)2013—2017年我國(guó)3家金7株PPRV流行毒株進(jìn)行血凝蛋白（H）基因序列測事哥定和生物信息學(xué)分析。對快結果顯示：37個毒株H基因核苷酸序列湖現之間的遺傳距離爲0~0.007 7，變異分布在41個位點，H蛋白氨麗們基酸序列之間的遺傳距離爲0~0.013 2，變異年南分布在24個位點。與15株代表毒株進章文(jìn)行序列比對(duì)發(fā)現，2013章近—2017年我國(guó)37株PPRV流行毒株H基因的13個位是睡點發(fā)生了核苷酸序列突變，其中9個導緻了氨基酸序列的改變。以最大似然法構遠照建分子進(jìn)化樹，發(fā)現2013—不人2017年我國(guó)流行的37個毒株構成(chéng)基因4系中一個獨我務立的進(jìn)化小分支。本研術生究闡明了2013—2017年我國(guó都樂)PPRV H基因的分子演化特征，從而爲該病控制去明和消滅策略的制定提供了數據支持。Molecular Evolution Ch友好aracteristics of 物自H Gene of Peste Des月女 Petits Ruminants Virus in是快 China During 2的坐013 to 2017In order to stu紙喝dy molecular characteristics飛國 of peste des petits ruminants viru對睡s（PPRV）in China，37 pre什銀valent strains of船下 PPRV collected from 2013 to 20在姐17 were used for h好舞emagglutinin（H）gene sequencin的東g and bioinforma信低tics analysis. The results showed that內刀 the genetic distance among nu愛年cleotide sequences of H 國服gene of these strains was 0 to 0.007放來 7，involving 41 variable sites，an也外d the genetic distance am飛懂ong amino acid sequences of H protein知煙 ranged from 0 to 0.公玩013 2，with 24 variable sites. Based on電議 the sequence alignment against 15得學 reference strains，it was found that 拿舊the mutation of nucleotide sequences 動市took place in 13 variable街什 sites of H gene of all these s海學trains，of which，9 nucleot放文ide mutations re商來sulted in the change of amino acid sequ厭照ence. In additi厭音on，a molecular evolutionary tree 紅會was constructed by maximum lik間醫elihood，it was found th藍弟at the 37 studied strains coul不說d be grouped into a distinct clade in l店體ineage IV. In short，the molecular evo愛訊lution characteristics of 務體H gene of PPRV in China from 2013 to 20費草17 were stated 大內in this paper，which would pr服河ovide data suppor的是ts to formulate relevant st雪議rategies for control and eradic高白ation of the disease.睡厭 全文下載鏈接：http://kns.cnk歌懂i.net/KCMS/detail/37.1246.S.20190903.1月美708.010.html
2019-09-17
熒光原位雜交在布魯氏菌和牛結核分枝杆刀妹菌檢測中的應用
熒光原位雜交技術可快速鑒定病原菌，較傳統病原分離鑒定方法具有明顯優勢。河答針對(duì)我國(guó)動物布魯氏菌病和牛結核病這(zhè)兩科關(liǎng)個重要的人獸共患病，目前還(hái)沒(méi)分服有标準的熒光原位雜交檢測方法可供參考門月。爲建立布魯氏菌和牛結核分枝杆菌熒光原位雜交檢測方法，快速診斷動物布魯氏菌病和拍黃牛結核病，利用布魯氏菌探針Bru-99問小6和結核分枝杆菌探針MTB770，通過務答(guò)優化雜交溫度、雜交時(sh機子í)間和樣(yàng)品處理等關鍵站放條件，确定最佳檢測程序；根據已知背景的菌株和臨床樣(yàng)品，對(duì現喝)Bru-996和MTB770探針的特異性和敏感性進(jì鄉視n)行評價，最終建立了熒光複位雜交診斷方法。結果顯技請示：反應條件優化後(hòu)，該方法可在窗新4 h内完成(chéng)布魯氏菌檢測；冷們熒光标記Bru-996探針與布魯氏菌待檢菌株的訊玩雜交結果均爲陽性，而與結核分枝杆菌、禽結核分枝杆菌和大腸杆菌雜交結果不就均爲陰性，并從5個已知背景的組織病料中成(chéng)功檢出布魯氏菌。也風牛結核分枝杆菌檢測則需要6~8 h，雜交前必須對(duì)樣(yàng)品用湖跳二甲苯和溶菌酶進(jìn)行處計紙理；MTB770探針可特異性識别并能(néng)從牛肺部結節中檢出牛結核分舊媽枝杆菌。結果表明，熒光原位雜交方法快速、簡便，而且Bru-996和MTB77花樂0探針分别在布魯氏菌和牛結核分枝輛司杆菌檢測上具有較高的特異性，可替代傳統的病原分離鑒定，作爲動物布魯路購氏菌病和牛結核病的實驗室确診方法。 App場睡lication of Flu筆街orescence in Situ Hybrid哥美ization in Detection of Brucella 水友and Mycobacterium b影站ovisCompared with traditional methods 劇男of pathogen isol去愛ation and identification水鐵，fluorescence in situ hybridiz為電ation（FISH）technology could be used to體來 rapidly identify pathogenic bacteria 制熱due to its obvious advantages. Howev輛校er，there has been no any f船好ormal FISH technology for two 科道kinds of major zoonoses（ani醫他mal brucellosis and bovin光來e tuberculosis）in China. In用船 order to develop the FISH technolo間議gy for rapidly diagnosing the two 道看diseases，the pr上畫obes of Brucella（Bru-996）and Mycob你拿acterium tuberculosis（MTB770）were used風跳 to determine the optimal detect睡輛ion procedure by optimizing the temp年會erature and time of hybridization請服，dealing with sam店笑ples and optimizing other important con相拿ditions. Then the spe人新cificity and sensitivity of the音師 two probes were evalu遠件ated based on strains with kn票綠own background a都厭nd clinical samples. The樂數 results showed河票 that Brucella c舊姐ould be detected wit遠我hin 4 hours after react見資ion conditions were optim這新ized. The hybridizat妹著ions between Bru-996 p文家robe and Brucella strains were positiv人物e，and that between Bru-996 probe and 她子Mycobacterium bovi會冷s，Mycobacterium avium and Esche畫土richia coli were all negative；and Br不能ucella was succe下公ssfully detected from five t頻制issue materials w廠區ith known background. It would也上 take 6 to 8 hours when 多說Mycobacterium bov雜數is was detected，and samples mus員睡t be treated with Xy要數lene and Lysozyme prior to hyb雜商ridization. Mycobacterium bovis技草 could be identified and detected麗員 from bovine pulmonary nodules b煙她y MTB770 probe. In vi個放ew of the advantages of FISH t分女echnology of rapidity and convenien文訊ce，and the high specificity of歌電 Bru-996 and MTB770 p林劇robes in detection of Brucella and Myc線弟obacterium bovis，i都站t was believed that the de刀市veloped FISH technology could r拍土eplace relevant traditiona得冷l methods and be used 姐家to diagnose ani放體mal brucellosis and bovine 自鐵tuberculosis in laboratories. 全文下載鏈接森到：http://kns.cnki.ne暗跳t/KCMS/detail/37.1246.S.2019日議0903.1708.032.html
2019-09-17
4株豬流行性腹瀉病毒的HA和HI試驗
爲觀察豬流行性腹瀉病毒（PDEV）的血凝麗他特性，進(jìn)而爲建立PEDV HA對得和HI試驗，評估疫苗免疫抗體水平奠定基礎，將(jiā爸事ng)CV777、DR13、YN13和書師YN144共4株PEDV毒株，在37 ℃下，分别用0、5、書醫10、50、100 μg/mL胰蛋白酶我黑處理30 min，然後(hòu)用雞、兔和豬紅錢小細胞進(jìn)行血凝（HA）試驗；用已經(jīng)鑒定土朋爲PEDV抗體陽性的血清和乳清分别進(玩但jìn)行血凝抑制（HI）試驗。音紙結果顯示：4株PEDV毒株隻與兔紅細一在胞發(fā)生凝集反應；CV777、DR章遠13毒株的HA現象需要用5 μg/mL的胰蛋白酶個吧進(jìn)行預處理，但是YN13和YN144毒株這子的HA現象不需要胰蛋白酶作用；該凝集現象能(néng)夠被(bèi)PEDV機這抗體特異性抑制。結果表明，PEDV能(néng)夠凝集兔藍金的紅細胞，不能(néng)凝集雞和豬數訊的紅細胞，因而可以用兔紅細胞建立PEDV的HA和愛身HI試驗方法。 HA-HI Test舊跳 for Four Strains of Porcine Epidemi著麗c Diarrhea VirusIn order to observe the商電 hemagglutination（HA）o鐘相f porcine epidemic dia視外rrhea virus（PDEV房年）and to lay a foundation for deve從生lopment of HA-HI te姐了st for PEDV and evaluati姐間on of the antibody le答高vel，four strains of PEDV，including CV海業777，DR13，YN13 and YN144，were tre站器ated with trypsin of 0，5，10，50 and民很 100 μg/mL respectively f請件or 30 minutes at 3開匠7 ℃，followed by HA test by use of re務我d blood cells（RBC）of chi用明cken，rabbit and pig respectively金站；then hemagglut報白ination inhibition（HI）test was c能如arried out with serum and whey that h很呢ad been identifi請土ed to be PEDV-po老通sitive. The results showed that fou我司r strains of PEDV only agglutinat睡拿ed with rabbit RBC. After CV777 子鐘and DR13 strain were pretreated with 5嗎熱 μg/mL trypsin，the HA phe的高nomenon occurred，while新又 no trypsin was 機員needed for YN13 andYN144 s員制train. In addition，HA c美會ould be specificall月校y inhibited by antibody 歌靜against PEDV. In conclusion，rabbit RBC 嗎電rather than chicken筆水 and swine RBC could be agglu討廠tinated by PEDV，therefore，the HA-HI te工船st for PEDV could be developed by use o農動f rabbit RBC. 全文下載鏈接：http://kns.工我cnki.net/KCMS/detai暗但l/37.1246.S.2019090商厭3.1708.036.html
2019-09-17
羊口瘡病毒B2L基因密碼子偏好(hǎo)性分析
B2L蛋白是羊口瘡病毒（ORFV）的主要囊膜蛋白，爲重要保護性抗原之門城一，可刺激機體産生強烈的抗體反應。本研究對(duì)B2L基因運用Vi媽海sual Gene Developer生物信息學(xué)軟件，分析你術其在大腸杆菌、畢赤酵母、昆蟲杆狀病毒表達系統中的密碼子使用偏好(hǎo)性，又線從而選擇一個最适表達系統，以增加B2L蛋白的表達量。分析表明，B2L那商基因在昆蟲杆狀病毒表達系統中會(hu拿算ì)得到更好(hǎo)的表達，這(術章zhè)爲B2L基因表達系統的選擇提供了參考。Analysis on t明她he Codon Preference of B2L Gene of Orf相哥 VirusAs the main envelope protei工要n of orf virus（ORFV），B2L protein答麗 isone of the important prot公工ective antigens銀務，which couldstimula現暗te the organism to produce 海家specific antibo森木dies. In the study，by use o紙道f bioinformatics software of Visual 學長Gene Developer，the codon preferen場子ce of B2L gene in the expressi的術on systems of E.coli，Pichia pastoris a中機nd insect baculovir現笑us wereanalyzed，and an optimal 舞湖expression system accordinglyw銀什as selected to i窗音ncrease the expr樹海ession of B2L protein. Accordi村風ng to theanalysis，B2L 習黑gene could be express低和ed better in the expression飛房 system of insectbaculovirus，which上我 would provide some references forsel下新ecting relevant expres說嗎sion system. 全文下載鏈接：http://上訊kns.cnki.net/KCMS/detail/37.1246.S但窗.20190802.1405.044.html
2019-08-13
