RNA病毒重組及發(fā)生機制
重組在RNA病毒中很常見，但有關重組的許多關鍵問題目前尚無法解釋。本文主要通去針對(duì)RNA重組的發(fā)生機制及其影響因素，解釋重組頻男跳率發(fā)生變化的原因，闡述RNA重組未來仍需解決的問題。RNA重組是河還RNA病毒特殊的遺傳信息交換方式，其頻率在不同的RN機黑A病毒中顯著不同。目前，研究者普遍認爲重組發(家體fā)生的機制可分爲複制型、非複制型重組以及重配，并森快且借助新方法和新技術，逐漸了解了影響該過(g睡月uò)程的病毒和細胞因子。有研究表明，R花舊NA病毒中RNA的重組和重配率非常高，這(zhè)與RNA依賴性RNA聚妹理合酶（RdRp）的保真性相關。因此，深湖東入了解病毒重組機制并發(fā)現其中的規雪答律，可實現對(duì)新型病毒以及新疫情的預警預報，從而有預錢師見性地控制疫病的發(fā)生和流行。R司的ecombination and the Mech你朋anism of RNA VirusesRecombination is w大科idely observed 關慢in RNA viruses，bu厭民t many key issues therein could人頻 not be explained ti金亮ll now. In the paper，the reasons f大不or the change of recombination fr家子equency were explained based on RNA r問妹ecombination mechanism and the infl很我uencing factors，and relevant issues to鐵雜 be addressed in the future were 電懂stated. RNA recombination was a通費 special way to費國 exchange genet月麗ic information by RNA v林海iruses，with different freq上工uency in various RNA viruses. At pr日日esent，it was generally believed by re花哥searchers that，the mechan著姐ism of recombinat下志ion could include repl新電ication，non-replication a窗媽nd reassortment，and the virus離生es and cytokines that might affec光雨t the process were g計樂radually recognized with t綠黃he help of new methods and technol書術ogies. It was studied that音地 the recombination and reassort分厭ment rate of RNA現習 were extremely hi船樂gh in RNA viruses，which was rela好大ted to the fidelity of dependent從開 RNA polymerase（RdRp到分）. Therefore，the new viru這近s or outbreak could be化鐘 early warned and r北明eported through further recognition of司低 the mechanism and rules of viru飛這s recombination，so as to control the oc海銀currence and prevalence 秒時of animal diseases樂多 in a predictable way.全文下載鏈南城接：https://kns.cnki.net/kcms/deta海看il/detail.aspx?d麗朋bcode=CJFD&dbname=CJFDAUTO&f長章ilename=ZGDW20210201時員8&v=2vpJqQNi66HOy5電舞JCBsWddjPx5O5qhK2daInInEe2DUKETJ鄉可Q0PrIidWOYKlD%25mmd2Fju4I
2021-02-26
鴨圓環病毒PCR檢測方法的建立與應用
鴨圓環病毒病是由鴨圓環病毒（duck circovirus，DuCV）引花行起(qǐ)的一種(zhǒng)重要免疫抑制病，影響著(飛身zhe)我國(guó)養鴨業的發(fā)展。爲建立針為你對(duì)DuCV的PCR快速檢測方法，對(duì)GenBank中服習登錄的DuCV全基因組序列進(jì秒下n)行遺傳進(jìn)化分析，通過(guò)對(duì)兩(liǎng)種的海(zhǒng)基因型的DuCV全基因組序列進(jì學好n)行比對(duì)，在共同保守區篩選出1對(duì)引物，建立了可以同時(s好商hí)檢測DuCV兩(liǎn外近g)種(zhǒng)基因型的PCR快錢少速檢測方法，并對(duì)該方法的敏感性哥能和特異性進(jìn)行評價。結對制果顯示，該方法具有良好(hǎo)的敏感性，檢測下限達理好1.0 fg；特異性良好(hǎo遠分)，對(duì)其他常見鴨病毒核酸檢測均爲陰性；利用該方法對請亮(duì)20份臨床發(fā)病鴨樣(yàng)品進(jìn)行檢測，發資事(fā)現有6份呈DuCV陽性多數，序列分析表明均屬于DuCV-1。該PCR檢測方法的建立爲鴨圓環病毒病的鐵都快速診斷和防控提供了重要手段。Deve東兵lopment and Application of a PCR A呢綠ssay for Duck CircovirusDuck circ睡兒ovirus（DuCV）disea樂作se is a serious immunosuppressive員家 disease caused by 器他DuCV，posing a t市雜hreat to the duck farming in China. In上睡 order to establish a rapid PCR ass訊我ay for DuCV，the geneti有來c evolution anal時明ysis was performed for the who飛報le genome sequen見遠ce of two DuCV genot樹關ypes that were registered in GenBank an秒對d subsequently compared with用吃 each other，then a pair of prim對森ers were screen懂窗ed in their common conservative ar舞能ea to establish a rapid PCR ass廠說ay that could detect t票下he two genotypes sim中西ultaneously. Then the sensi議土tivity and specificity of the me用光thod were evaluated.風資 The results showed th兵低at，by the method，the lower 外紙detection limit could 科玩reach 1.0 fg due to 鐘民its good sensitivity；all nucl懂裡eic acid detection for othe錢花r common duck viruses were negativ但道e due to its good specificit海腦y；6 out of 20 samples of clini知白cally infected ducks were posit北紅ive for DuCV，which fell into D服知UCV-1 as indicated by sequence analys女土is. In conclusion，the method 厭計could rapidly diagnos爸房e，prevent and control DuCV as an impor資做tant tool.全文下載鏈接：https://kns.cnki窗土.net/kcms/detail/detail.aspx?dbc校計ode=CJFD&dbname=朋刀CJFDAUTO&filename=關現ZGDW202101021&v=2vpJqQNi地下66GwD6p6B%25mmd2金靜FdPDgcZ9km9zF8R7QPpJ2wJj%25m理物md2BAqhBLbS7YWcN4DvUX%25mm家如d2FKpLt
2021-01-14
鑒别H5亞型兩(liǎng)種(zhǒng鐘拿)分支禽流感病毒雙重實時(shí)熒光RT-PCR方法的建立及應用
爲準确區分H5亞型禽流感病毒2.藍紅3.2.1分支和2.3.4.4分支的毒株，通過(gu得音ò)對(duì)GenBank和GISAID數據庫中發(fā)表草工的以及本實驗室保存的H5亞型禽流了視感病毒HA基因序列進(jìn)行比對關計(duì)，設計1對(duì)特相玩異性引物和2條探針，建立了區分H5亞型禽流感病毒不同鄉術分支毒株的實時(shí)熒光RT-PC日農R方法，并對(duì)該方法的反應體系和反應參數進(jìn)行了優化微問，同時(shí)開(kāi)展了特異性和敏感性試驗，以及臨床應用檢測。結如聽果顯示：該方法具有良好(hǎo)的特異性，與其他亞子場型禽流感病毒及常見禽病病毒無交叉反應；對(duì)H5亞型禽流感病毒2.3.2長學.1分支和2.3.4.4分支毒株的檢測靈敏度分别爲49錯愛5.0 copies/μL和22.市火3 copies/μL；100份臨床家禽拭子禽流感病毒檢測結果與常規RT木白-PCR和病毒分離檢測結果一緻，符合率爲100%。結果表明，該方法特異、他大敏感、準确、耗時(shí)少，同時(shí)還(hái)可區分不務話同分支，可應用于H5亞型禽流感理跳病毒的準确、快速檢測。Establishment and 地些Application of a Dupl話看ex Real-time RT-PCR Assay for Identi對林fying Two Branches of H5 Su就麗btype Avian Influenza Vir訊話usIn order to accurately di裡鐵stinguish the strains時都 of two evolutionary clades inc水大luding 2.3.2.1 and 2.3.4.4到廠 of H5 subtype avian in厭學fluenza virus（AIV），a pair of 如朋specific primers and two TaqMan去新 probes were designed by comparing HA g下鄉ene sequences of H5 subt習門ype AIV registered時弟 in Genbank and GISAID wi內也th the ones reserved相匠 in our laboratory，a dupl河相ex real-time RT-PCR assay 從輛was established to dis哥費tinguish different clades of th妹計e virus，and its reaction錯物 system and param好喝eters were optimized. Meanwh慢視ile，the specific到業ity，sensitivity and its民不 clinical appli這去cation were tested. The results sho也場wed that，the specificity of訊見 the method was good 線那with no any cross-reaction wit會請h other subtypes AIV or 畫會common avian viruses；the detection 海朋limits of clades 2.3.2.1 and 2體很.3.4.4 were 495.0 and 22.技見3 copies/μL，respective術讀ly；100 clinical 影人poultry swab samples were tes我河ted by the established m東少ethod，and the results were consis的還tent with those by traditiona的嗎l RT-PCR and virus is到短olation，with the coincidence rate of 1技謝00%. As a conclusion，the e男還stablished method was speci山國fic，sensitive，accurate and conve日你nient，which could be u校放sed to distinguish different clades新雨 of H5 subtype AI中朋V.全文下載鏈接：https://kns.cnki.net/kc坐動ms/detail/detail.aspx?又弟dbcode=CJFD&dbname=CJFDAUTO&file在去name=ZGDW202101020&厭個v=2vpJqQNi66HpC2DtPpxmZ7MbcnGvT1y1c上匠aIfZvmjLYrGOkLUTMOCulgxxUIz4kWz科開
2021-01-14
牛源多殺性巴氏杆菌PlpE蛋白的動北原核表達及多克隆抗體制備
爲體外表達牛源多殺性巴氏杆菌（Pasteurella錯拍 multocida）PlpE基因及制備抗PlpE蛋白多克隆抗體吃河，根據PlpE基因核酸序列設計了1對(duì)特異性引物，通過(票雪guò)PCR擴增PlpE基因，構建重組質粒pET-32a-PlpE；搖商西菌提取質粒，進(jìn)行雙酶切鑒定，鑒定正确後(hòu)將(樹醫jiāng)重組質粒pET-32a-PlpE轉化地房至BL21（DE3）感受态細胞，加入IPTG進術木(jìn)行誘導，對(duì)誘導蛋白進(計女jìn)行SDS-PAGE和Western Blot鑒定；使用純化蛋白訊學PlpE免疫北京大耳白兔，制備多克隆抗體，并進(jìn)行能錢間接ELISA和Western Blot分析。結果顯示，擴增的PlpE基因大藍湖小爲1 050 bp，在原核細胞中誘導表達的森森PlpE蛋白約56 kDa。間接ELISA結果顯示，所制備多克些還隆抗體效價可達1:512 000；Wester湖好n Blot結果顯示，所制備多克隆抗體反應性良好(hǎo)。PlpE蛋白的成(從頻chéng)功表達和多克隆抗體的成(白說chéng)功制備，爲下一步多殺性巴氏杆菌診斷方法的建務關立和疫苗研發(fā)奠定了基礎。Proka美就ryotic Expression o小物f PlpE Protein of Bovine Pasteurella mu低購ltocida and Preparation of Po一筆lyclonal AntibodiesIn order to ex大高press the PlpE gene of bovi生城ne Pasteurella multocida in vitro和會 and prepare cor白看responding polyclonal antibodies，微國a pair of speci金信fic primers were designed according 在務to the nucleic acid sequence of話長 the gene，then pE友小T-32a-PlpE，the reco姐算mbinant plasmid，was constructed through購了 PCR amplification，after plasmid e文自xtraction and identification by金可 double enzyme digestion好那，the recombinant謝路 plasmid was tr票吃ansformed into BL21（DE3）ce白習lls and the bacteria was indu湖這ced by IPTG and the induced prote在小in was identified by SDS-PAGE and區微 Western Blot；Beijing White Rabbi理遠ts were vaccinated with 雜線purified PlpE p北作rotein to prepare polyclonal anti什做bodies，which were analyzed by你水 indirect ELISA and Western Blot.了們 The results showed that th站件e amplified PlpE gene火見 was 1 050 bp，and上知 the induced PlpE protein wa知新s about 56 kDa. It was shown that，b答習y indirect ELISA，the ti靜新ter of the polyclonal ant火又ibodies could be up to 報能1:512 000；and by Western Blot，the anti紅空body could react wel中問l with the positive serum. It was爸男 concluded that a foundation was現是 laid for the establishment o內低f the method for identify體會ing Pasteurella mul民窗tocida and development of relevant 船年vaccines in the future.全文下載鏈接：http線房s://kns.cnki.net/k樹到cms/detail/detail.aspx?db新雨code=CJFD&dbname=CJ術兵FDAUTO&filename=電事ZGDW202101019&v=2vpJqQNi66FspGXrYE哥這bDEJvrXEmDYO9pI6yc9rrux3Q6YI6dTx醫會xTKTZwNyU3xVug
2021-01-14
微流控芯片技術在微生物檢測中的應用
微流控芯片是一項以芯片爲基礎，結合化學(xu吧森é)、物理學(xué)、生物學(xué)等多學(x信你ué)科形成(chéng)的技術平台，能(néng)夠在員高一個芯片上實現樣(yàng)品制備、朋放反應、分離、檢測等流程，具有高通量、高效率和易操作了姐的特點。随著(zhe)材料科學(xué)和納米技術的發(f微中ā)展，該項技術得到迅速發(fā)展并應用如一于化學(xué)分析、微生物檢測等，尤其是在微生物檢測和分析方面(miàn市錢)發(fā)揮著(zhe)重要作用，并逐步産生時花了商品化的微流控芯片。微流控芯片因具有小型化、節約試劑、速度快、集成(c業高héng)化程度高等優點，适用于處理突發(fā)公共衛生事(shì)但農件，但目前在動物疫病檢測中的應用還(hái)很少見。本文對(d讀視uì)微流控芯片的技術原理、結構及其在微生物檢測中的應作坐用進(jìn)行綜述，以期爲動物疫病的高通量檢測提供借鑒。A姐我pplication of Micro錯去fluidic Chip Technology in M生鄉icrobial DetectionMicrofl員弟uidic chip is a kind of technol開視ogy platform ba山開sed on a chip，which combines with sever長費al disciplines，such as chemistry，家場physics，biology and others，and could co身媽mplete the processes of 購和preparation，reaction，s雨文eparation and d看好etection of samples on a chip due to 河朋its characteris分爸tics of high throughput，hig年知h efficiency and easy operation. With 又土the development o開朋f material science and nanotechno美車logy，the technology的北 has been rapidly deve行慢loped and applied to chemical analys行來is，microbial detection and又暗 so on，in which，it has been pl就做aying an important role in mic業匠robial detection an草為d analysis，and gradually leads to t舞現he commercial microfluidic chip that 站通is advantaged with mi文也niaturization，reagent 冷理saving，fast and high i對和ntegration，and appropriate for dea藍亮ling with public health emergencies，b醫就ut is still seldom used 志時for animal disea算能se detection at prese木南nt. In the paper，the technical pri媽理nciples and structure of microfl生舊uidic chip as well as its applica商又tion in microbial det可村ection were reviewed，with 行門a view to providing some 討服references for high-t兒綠hroughput detecti玩兒on of animal diseases.全文下載鏈接：h錢小ttps://kns.cnki.net/kcms/detail/deta都國il.aspx?dbcode=CJFD&dbname=CJFDAUTO爸海&filename=ZGDW202101018&v=2vpJq民制QNi66FIJLK2OSEL就年8Yi4RRdCUFoM1KEDG子相CyKgnlUlreyqw0sri%25mmd2Bh街們1Vrcaqxh
2021-01-14
豬繁殖與呼吸綜合征檢測技術與疫苗研發(fā)進(jìn)展
豬繁殖與呼吸綜合征（PRRS）給世界養豬生産造成(chéng)了巨大經劇自(jīng)濟損失。對(duì)PRRS的精準檢測以及疫苗的科自家學(xué)使用是臨床PRRS防控的重中之重。爲此，本文綜述了PRRS檢測技計外術和疫苗研發(fā)進(jìn)展，以期爲臨床PRRS綜合防控提供著車依據。目前臨床上檢測PRRS的主要技術包括血清抗體大綠ELISA檢測以及各種(zhǒng)類型的RT朋看-PCR檢測技術。ELISA檢測以評價服匠疫苗免疫效果或野毒感染情況爲主要目的，RT-PCR檢測區見以臨床診斷及調查野毒株類型爲主要目的。在疫苗方訊費面(miàn)，目前市場上主要有活疫苗、滅活苗靜可、亞單位疫苗、DNA疫苗以及載體疫苗等不同類答拿型的單苗以及聯苗。其中：活疫苗保護效果較好(h自現ǎo)，但交叉保護不強；滅活苗安全性較高，但免疫效力較差；亞單位疫一報苗可用于妊娠晚期；DNA疫苗可增花房強活疫苗的保護效果；載體疫苗雖具有部分保護力，但未進(jìn)行實際為離應用。目前依舊沒(méi)有完全符合國(guó)際新一代件用标準的PRRS疫苗上市。因此，通過(guò)科學(xué)檢測影線以及將(jiāng)生物安全和合理的疫苗接種(zhǒng)結合起(qǐ)來林一，進(jìn)行田間毒株的檢測和基因分析事鄉，進(jìn)而選擇相應基因型毒株疫苗進(jì樹懂n)行免疫，對(duì)控制農場以及地區的PRRS具有非常重要的作用。R來紙esearch Progress of the Diagnos錢也tic Techniques and Vaccine有道s for PRRSPorcine repro老他ductive and respiratory syndrom月雜e（PRRS）has brought huge economic lo湖雪sses to pig produ費那ction in the world，which should 技門be prevented and controll人事ed with the top priority議匠 of accurate de如風tection and scient現用ific application of vaccines. At p有兒resent，the main technique在亮s to diagnose PRRS i數妹ncluded serum antibody ELISA assay an文能d various RT-PCR師舊 assays. The for文妹mer aims to evaluate the雪從 immune effect of vaccines o文子r wild virus infection，while the l月報atter focuses on cli說些nical diagnosis and investi門弟gation of wild virus strains. For vacci照可nes，all kinds of single街河 and combined vaccines are available i呢視n markets，including live vaccines，inact拿煙ivated vaccines，subunit vaccines，DNA 他國vaccines，vector兵也 vaccines，etc.，specif光知ically，live vaccine問都s are with good effect but poor cro靜爸ss protective immunity；inacti吧作vated vaccines are with森河 high safety but poor immune 月他effect；subunit 畫來vaccines can be used in late pregn請計ancy；DNA vaccines can increas美還e the protection effect of live vacc大工ines；and vector vacc鄉視ines have not be西國en applied in practice although the公費y are with partial protective immu妹自nity. No any vaccines against PRR視那S that fully meet the n匠又ew international standard聽下s have been available till做為 now. Therefore，it will be v南樹ery crucial to control P科湖RRS in farms or r車河egions through scien時公tific diagnosis and combination跳學 of bio-safety and reasonable vaccinati工他on to detect field strains 文新and analyze their genes，and t知站hen to vaccinate wi物弟th corresponding vaccines.全文下載鏈朋看接：https://kns.cnki.net/kcms/光相detail/detail.asp拿喝x?dbcode=CJFD&dbname=CJFD農煙AUTO&filename=ZGD家了W202101016&v=2vpJqQNi美爸66ErxHSyLOD3MXoXltAc3QuZhVZLG5Vb技師gKZIUFf2BdT1YOqRKWETMBD8
2021-01-14
基于網絡藥理學(xué)分析黃芪防控豬繁殖與呼吸綜合征錯草的物質基礎及靶點信息
爲揭示黃芪防控豬繁殖與呼吸綜合征草頻（PRRS）的物質基礎及靶點信息，采用網絡行上藥理學(xué)方法，從中藥系統藥理學(xué)數據庫與分析平台（TCM頻拿SP）篩選黃芪的潛在活性成(ché街來ng)分、作用靶點，同時(shí)從比較毒理基因組學(xu麗高é)數據庫（CTD）收集PRRS相關宿主基因，最後林睡(hòu)構建“活性成(chéng)日離分-關鍵靶點”網絡、蛋白互作網絡，并進(jìn)行GO功能他們(néng)和KEGG通路富集分析內林。結果顯示，從黃芪篩選出的17個潛在活性成(chéng湖做)分中，有14個潛在活性成(ché近風ng)分共計65個基因靶點與PRRS相關宿主基因交叉子嗎，其中槲皮素、山奈酚、刺芒柄花素、異鼠李素等煙聽潛在活性成(chéng)分可能(néng)對(duì)治療PRRS有我務重要作用，TNF、IL8、TP53訊弟、IL6、IL10、IL1B、JUN等45個蛋白也影靶點間存在相互作用。此外，黃芪治療PRRS的作用機制可能(民得néng)涉及細胞外間隙和胞漿等組分，光林并可能(néng)與炎症反應、細胞凋亡呢金、T細胞受體信号通路等生物過(guò)程相關。本研究爲臨床應用黃芪及其提取新會物治療PRRS奠定了理論基礎。Analysis on the Mate書花rial Basis and Drug Targets做自 of Astragalus Membranaceus in Pre用路vention and Control o銀森f PRRS Based on身雪 Network PharmacologyIn民要 order to reveal the informatio會水n concerning the material吃線 basis and drug t船地argets of astragalus 鐵嗎membranaceus acting on porcine reprod少報uctive and respiratory syndro城但me（PRRS），the potential activ老訊e components and drug targets relate老中d to astragalus membranaceus 說上were screened through tradi也少tional chinese medicine 兒兵systems pharmacology databa外腦se and analysis platform森日（TCMSP）by the network pharm身站acology. Meanwhile綠兒，the host genes時河 related to PRRS were gat拍票hered from comparativ放村e toxicogenomics database（CTD）to constr個有uct the“active compone電可nts-key targets”network and protein開個-protein interaction（PPI）ne店農twork，and to analyze the GO f上用unctions and KEGG pathway e下嗎nrichment. The results show自現ed that 14 out of黑紙 17 potential active輛廠 components including 65 gene t站會argets crossed with the host genes rel音作ated to PRRS，esp秒南ecially，quercetin，kaempf站美erol，formononetin，isorhamnetin and計還 other potential activ就技e components played an i頻報mportant role in fighting agai從物nst PRRS，and 45 targets could interac關風t with each other，including TNF，但著IL8，TP53，IL6，IL1遠算0，IL1B and JUN. In addition，the行路 mechanism of ast林河ragalus membranaceus in fi門弟ghting against PRRS might invo影要lve extracellul會數ar space and cytoplasmic c高雨omponents，and might be r短銀elated to biological processes such as 資要inflammatory response，apop志市tosis，and T cell rec書可eptor signaling pathway. 睡白Therefore，a theoreti醫民cal foundation was l相在aid for clinical application of astrag村錯alus membranaceus做分 and its extract 到章in fighting against PRRS. 我有全文下載鏈接：https://kns.cnki.net/kcms/detai機問l/detail.aspx?dbcod計也e=CJFD&dbname=CJFDAUTO&fi看民lename=ZGDW202101015&v=2我信vpJqQNi66FXF9R5APrxlb笑哥EWzpYeoKIo%25mmd2Fr歌麗CDWaNAQ5qxqiEskBfDFtKQggeuabBJ
2021-01-14
鴨坦布蘇病毒山東流行株的分離鑒定及其基因組序列分析
爲了解山東省鴨坦布蘇病毒（du區村ck tembusu virus，DTM弟資UV）的流行與變異情況，對(duì)2018年身要下半年至2019年上半年泰安、濟甯和淄博等地黑又送檢的臨床病鴨樣(yàng)品進(jìn)行了DTMU內就V病原學(xué)檢測，并對(duì)陽性病料進(jìn)行病毒分鐘朋離鑒定及分離株的全基因組序列分析。結果顯示：從1風冷02份樣(yàng)品中，共檢測到8份DTMUV陽性，陽性麗間率檢出率爲7.8%，從陽性病料中分離到3株DTMUV分離株；3個分都費離株基因全長(cháng)均爲從有10 993 bp，彼此間的氨基酸同源性爲99.2%~姐爸99.3%，與24個參考毒株的同源性爲95.8%~99城玩.9%，均屬于中國(guó)株I型；與參考毒株吧飛相比，分離株的核衣殼蛋白C、囊膜蛋照我白E和非結構蛋白NS1的同源性相對(duì)較低員秒，分别爲86.1%~99.2%，海花95.0%~99.4%和92.3%~錢還99.7%，同時(shí)存在數量女女不等的氨基酸突變位點，但關鍵的拍懂蛋白結構和潛在的糖基化位點無差異。結果表明你子，DTMUV在山東部分地區仍有流行，但流睡農行毒株基因結構及緻病特性與經(j工在īng)典毒株相比無明顯變化，但應愛嗎持續關注和深入研究。Isolation and 如紙Identification of Shandong Str我和ain of Duck Tembusu Vir相內usIn order to iden匠員tify the prevalen懂男ce status and variation上校 of duck Tembusu virus（DTMUV拿南）in Shandong Province，the雜門 clinical samples delivered from T但和ai'an，Jining and Zibo during t說算he second half of 2018 to不還 the first half of 高鐘2019 were detected for DTMUV，and南鄉 virus isolation and identificat家我ion were carried out tow遠風ards the positive samples，then the who銀你le genome sequences of the isolates wer火商e analyzed. The results showed that 8離拿 out of 102 samp唱說les were positive，with the d短是etection rate of 7.8%，from which，3 stra林得ins of DTMUV were i湖黃solated；the full length錯照s of all the three strains靜刀 were all 10 993 bp，the amino acid homo放房logy of they ranged f中暗rom 99.2% to 99.3%，and the homolo店工gy between the isolat費劇es and 24 reference strains ranged fr林少om 95.8% to 99.9%，all醫都 the isolates fell into Chinese 些金strain I；compared to the reference s花銀trains，the homologies o下生f nucleocapsid pr樂鄉otein C，envelope protein 民著E and non-structural protei道國n NS1 of the isolates were lowe分錯r，which were 86.1% to 9服北9.2%，95% to 99.4% and 92.3% t紅喝o 99.7%，respectively，舞雪and with different num雜個ber of amino acid mu海拍tation sites，but majo議有r protein structures and potenti地飛al glycosylation sites had no 科土difference. It was concluded 電她that DTMUV was still prevalen請件t in some regions in Shandong 了水Province，which should be further conce少商rned and studied，although its g體爸enetic structure and pat又報hogenicity remained 歌明unchanged compared to the笑還 classical strains.全文下載鏈接：http舞光s://kns.cnki.net/kcms/detail/de筆從tail.aspx?dbcode=話新CJFD&dbname=CJFDAUTO&filename=ZG暗東DW202101008&v=2vpJqQNi66H技飛AmotFUcqF5c8IG0hCBj%25mmd2F機拿jC6JPalOXfdjJ1WIX49lsO7qv2土生5k1hfwp
2021-01-14
安徽省6個地市規模雞場新城疫流行率估計及風險因素分析
爲了解安徽省規模雞場新城疫病毒感染水工情況及其引發(fā)該病的風險因素，結合2019年秋季集中監測采樣(yà去多ng)工作，對(duì)6個地市存欄3 000~10 000隻的規模也間雞場開(kāi)展橫斷面(miàn)研究。采用兩(liǎ北些ng)階段随機抽樣(yàng)，在6個地市範圍内，随機抽取153個規模雞場筆可，采集5 369份咽喉/洩殖腔拭子采用熒光PCR進短家(jìn)行病原學(xué)檢測，同時(筆討shí)進(jìn)行問卷調查，分析城去相關風險因素。結果顯示，6個地市153黃雨個規模雞場的場表觀流行率和場真筆村實流行率分别爲11.1%和11.7%。單因素分析媽匠顯示：“1個月隻清理1次糞便”和“場内共用器具、車輛”是潛在風險因素，有統計媽靜學(xué)意義（P＜0.05）；車文聽輛和人員進(jìn)出場消毒、淨道(dào)和吃都污道(dào)分開(kāi)、空舍消毒得畫以及消毒前清洗場内可以降低感染風險。Logistic回歸分析來湖顯示，“淨道(dào)、污道(dào)不分開(kāi)”和“車輛、人員進(著器jìn)出不消毒”是風險因素。結果表明，安徽省部分雞場仍存在新城動空疫病毒感染，應加強雞場生物安全管理，合理規劃養殖場布局，白上場區入口要建造消毒池，場内淨道(dào)和污道(家花dào)要分開(kāi)，車輛和人員進(jìn)出場時(shí)要還信嚴格消毒，場内要制定詳細的消毒計劃，切實改善養殖場環境。Estim雪錢ation of Newcastle Disease P筆請revalence in Scale Farms in 6 Cities of為了 Anhui Province and Anal懂制ysis on Relevant Risk Fact唱知orsIn order to identify the infe嗎雨ction status of Newcastl音錯e disease virus（NDV）and ris火機k factors leadin森亮g to the disease in A工離nhui Province，an cross-sectio科信nal study was carried out in th錢紅e scale farms with an雨就 inventory of 3 000 to答哥 10 000 chickens in six cities in co討愛mbination with centralized su在中rveillance and sampling金業 in the fall of 2019. A total 事劇of 5 369 throat/cloac外內al swabs were randomly collected 呢裡from 153 farms by two-stage sampling湖呢 method. Meanwh房務ile，a questionnaire investigation wa工們s conducted to analyze 器票relevant risk factors. The月現 results showed th劇商at the apparent prevalence and t腦說rue prevalence were 11.1% an錯醫d 11.7% respectively at t唱微he farm level. It was shown 熱煙that，by monofactor an慢看alysis，the poten請笑tial risk factors incl都內uded“only cleaning feces once a m化討onth，sharing equipment and 作現vehicles within the farms”，which厭外 was of statistical signi一靜ficance（P＜0.05）；a知愛nd the risk of infect術風ion could be reduced by麗低 disinfecting vehicles and personnel wh算師en they were entering or在刀 leaving farms，separating clean cha日街nnels from the 分個dirty ones，disinfecting empty pens an吃關d cleaning the 自大premises prior to disinfecting. It wa和高s shown by Logistic regression analysis醫土 that，the risk factors incl雪南uded“failure to separate clean風遠 and dirty channels and 兵通to disinfect vehicles and per個森sonnel in or out of farms”. As a concl厭吃usion，NDV was still pr的好evalent in some farms in Anhui商水 Province，and some suggestions were 學用put forward，such as 務水strengthening bio-security management請兒 in the farms，rea白新sonably designing動錯 their layout，building a 校來disinfection pool at the entr就讀ance，separating clean chann司要els from the dirty ones，月東strictly disinfecting vehicles and pers來店onnel when they 身和were entering or leaving farms，and dev姐暗eloping detailed disinfe山理ction plan to practically improve 那媽the environment of the far們服ms.全文下載鏈接：https://kn都她s.cnki.net/kcms但技/detail/detail.asp知國x?dbcode=CJFD&dbname=CJFDAU事懂TO&filename=ZGDW202101006&v=2vpJqQNi66H東厭aOQK62RPPfwn8x6iknDBctOCeF窗房2DdBBqt7ZnZcqNDxFfJKn3aOdPd
2021-01-14
羊駝免疫庫的制備及抗孔雀石綠納米抗體的篩選
孔雀石綠（malachite green，MG）是一一校種(zhǒng)易溶于水和乙醇等的基礎染料，可在多種(zhǒng)行業應用志風，并曾用于水生動物多種(zhǒ風業ng)疾病如真菌病、寄生蟲病、細菌性疾病等的知服治療。但因其具有緻癌等毒副作用，目前已被(bè厭務i)禁用。然而市場上仍有在水産品違法使用MG的現象，因此廠黃需要加強對(duì)該藥品殘留的檢測。刀水本研究選擇羊駝免疫庫篩選抗MG納米抗體，通過(guò)噬菌體ELISA篩選高親年討和力和特異性的納米抗體。通過(guò日慢)輔助噬菌體M13的救援，使得納米抗體基因（VHH體理）與噬菌體外殼蛋白基因融合表達，從而使得VHH蛋白展示在噬菌體表男腦面(miàn)，通過(guò)有限稀釋購畫篩選和多次洗脫的辦法，獲得含有目的又店基因的單個噬菌克隆；將(jiāng)重組質粒PMES4-VHH轉化到表達影我細胞中，進(jìn)行體外表達。最後(hòu)采用NI-NT謝他A柱進(jìn)行His标簽純化科話，得到抗MG納米抗體，并進(jìn)行SDS-PAGE電泳鑒定。結果顯示：答筆噬菌體文庫成(chéng)功構建，庫個醫容爲3.2×108 cfu/mL；通過(g你家uò)ELISA篩選，得到16個具有抑制效果的噬菌體克隆，經(jīng)對公就(duì)噬菌體基因測序并翻譯，得到2種(可明zhǒng)氨基酸序列；將(jiāng)重組質粒轉化至表達了輛菌株表達、純化，最後(hòu)得到大小約爲1厭身5 kDa的抗MG納米抗體，從而爲MG的下一步檢測奠定了朋動基礎。Preparation of Alpaca Immune Rep他個ertoire and Selec市通tion of Nano-antibody against Malachite遠哥 GreenMalachite green（MG）is a kind 明上of primary dye that is solubl樂畫e in water and ethanol，村他and has been ap線睡plied in many indust空木ries，especially to fight against 知藍many diseases of aquatic animals，河年such as fungal diseases少年，parasitic diseas聽內es and bacterial diseas笑友es. But it has been bann大機ed due to its adverse reaction such 頻友as carcinogenicity北愛. However，it is necessary to strengt筆物hen the detectio拿務n of drug residues from MG t可黃hat is still illegally是他 used in aquatic products in mar舞視kets. In this study，nan木銀o-antibody with high affinity and spec呢她ificity against MG w愛很as selected from al舊他paca immune repertoire through少學 the phage by ELISA，then科站 was used to support the rescu但光e of phage M13，so th文得at nano-antibody gen樂間e（VHH）could be fused and expressed章都 with the coat protein gene of the化見 phage，and VHH protein 秒城could be shown on the surface of the ph校拿age to obtain the single phagocy可北tic colonies containing the低東 targeting gene by means of limited di到西lution screening and multipl得很e elution；the recomb雜樹inant plasmid PMES4-VHH were 又我transformed into exp黃得ression cells and expressed in農窗 vitro，then His-tag purification was 森從carried out by the NI-NTA colu體弟mn to obtain the nan日去o-antibody against MG that wa大鐵s identified by SDS-PAGE electroph紅身oresis. It was 體會concluded that the immune 我新repertoire was su麗就ccessfully constructed 好熱with a capacity of 3.2×108 cfu/m還拿L；16 colonies with inhibitory 照煙effect were scre開內ened by ELISA，two kinds of amino acid 跳藍sequences were obtained討腦 by sequencing and tra匠行nslating the pha服身ge genes；PMES4-VHH was transformed i小如nto the expression 些線strain for expression and purific光對ation，and the nano-ant他綠ibody of about 15 kDa agai媽金nst MG was produced，which paved the wa兵都y to future detect和雨ion of MG.全文下載鏈接：https://kns.cnki.net員坐/kcms/detail/deta月唱il.aspx?dbcode=CJFD&dbname=CJFDA風大UTODAY&filename=ZGDW202012鐵場025&v=Bh8a8ZT44放林%25mmd2BXk0ZJ7KNY熱低HWtHzXPUmzT5DElVbdXML%25mmd2FMLk8l%25m她東md2FMwEnYZUQ9XabHs0ZU
2020-12-03
應用豬繁殖與呼吸綜合征核酸标準物質比較5種(zhǒng)核酸提取試劑盒
爲比較不同核酸提取試劑盒對(du但國ì)豬繁殖與呼吸綜合征病毒（PRRSV）核酸的提取效率，使用國(guó商火)家标準物質“豬繁殖與呼吸綜合征病毒美洲經(j錯照īng)典株核酸标準物質”作爲模拟樣(yà不有ng)品，分别使用試劑盒A、B、C（磁珠法）以及員又試劑盒D、E（柱式法）等5種(zhǒng)志草核酸提取試劑盒進(jìn)行核酸提取，然後(hòu)通過(guò)熒光RT-多子PCR和數字RT-PCR方法評估提取效率，并分析不同基質對區湖(duì)提取效率的影響。結果顯示兒路：5種(zhǒng)試劑盒均能(néng)有效提取PRRSV RNA，其中磁珠謝器法的提取效率高于柱式法；試劑盒B和C對(玩拍duì)PRRSV RNA的提紙謝取效率較高；基質會(huì)對(duì弟熱)提取效率造成(chéng)影響，精液和醫生抗凝血對(duì)試劑盒提取效率的影響較大。綜上，試劑盒B、C均可适用于豬還東繁殖與呼吸綜合征臨床樣(yàng)品的核酸提取。實際會睡操作中，應根據實驗室條件、樣(yàng)品數量、樣(yàng)品類型技為和時(shí)間要求等因素，選擇應用合适的提取試劑盒。Comparison畫費 of Five Kinds of Nucleic Acid Extracti女腦on Kits Based o和亮n Nucleic Acid Ref事很erence Materials of PRRSVIn or志火der to compare the efficiency o身知f various nucleic acid extraction kit雜行s for RNA of porcine reprodu花黃ctive and respiratory syndrome 畫件virus（PRRSV），national reference mate近議rial“PRRSV American 小明classic strain nucl樹輛eic acid reference material”was知老 respectively extracted by five 要議kinds of kits（three magnetic bead ki站工ts including A，B and C and two 微化column kits includi哥司ng D and E），and their動藍 efficiency were 事暗evaluated by fluo說好rescent RT-PCR and digital RT-PCR. 信員Then，the effect of differ答厭ent substrates on th朋話e efficiency was a自數nalyzed. The results showe現吃d that all the k火靜its could effectively extract RNA of 做票PRRSV，in which，the efficiency of magne火動tic bead kits was higher than that 廠是of column kits；the efficiency of B an遠白d C was higher；the efficiency美黑 could be affected by substrates，esp得筆ecially semen a去訊nd anticoagulant. In conclusio去放n，both B and C 中工were appropriate for extraction of nucl放就eic acids from clinical samples志件 of PRRSV. Therefore，ap笑說propriate kits should 電南be used subject to la科草boratory conditions，number of sampl司事es，types of samples，urgency of ca理南ses and other factors.全文下載鏈接：空數https://kns.cnki.net/kcms/detail/detail作外.aspx?dbcode=CJFD&dbname=CJ跳冷FDAUTODAY&filename=ZGDW2020120車街23&v=Bh8a8ZT44%2多一5mmd2BVdUZJJ4fPLG4mzwxK%25mmd2Fy舞知yV5Gx5EoETBAWZubmLyLyQ現樹6p%25mmd2B%25mmd2BCT0seR3媽子R9
2020-12-03
A型塞内卡病毒VP1結構蛋白在昆蟲杆狀病毒中的表達與鑒定
目前在昆蟲杆狀病毒表達系統（baculo房又virus expression vector sy聽你stem，BEVS）中表達A型塞内卡病毒（S上線enecavirus A，SVA）結構蛋白V黃中P1尚未見報道(dào)。本研究將(jiāng)電刀VP1基因通過(guò)限制性酶切位點插入杆狀病毒載體pFastB技從ac I，然後(hòu)將(jiāng)鑒定正确的重組載體轉化至DH1學個0Bac感受态細胞，經(jīng)藍白斑篩選，對(duì)鑒定正确的菌遠在落提取質粒。將(jiāng)質粒轉染Sf9昆日視蟲細胞，待有明顯細胞病變後(hòu)謝長收獲重組杆狀病毒rBac-I-VP1，并進(腦現jìn)行病毒滴度測定。随後(hòu)通過(guò)土能蛋白免疫印記（Western Blot）和間接免疫熒訊街光（IFA）鑒定VP1蛋白表達情況。結個朋果顯示，重組杆狀病毒rBac-I-VP1滴度爲6.8×105 pfu/mL；人媽Western Blot可檢測從是到相對(duì)分子質量約27 kDa的表達産物，家店且其能(néng)夠被(bèi)S姐開VA兔陽性多克隆血清識别；IF少黑A試驗顯示，VP1蛋白能(né厭低ng)夠在Sf9細胞内表達。結果表一小明，本研究利用BEVS表達的SVA 爸下VP1蛋白具有良好(hǎo)的免疫反應性，爲其後(hòu)期功能(né從老ng)研究及SVA診斷試劑研制提供了技術基礎。Ex文們pression and Identifi靜上cation of Senecavirus A到技 Structural Protein VP1 in I體南nsect Baculovir花民us The expression of現兒 senecavirus A（SVA）structural拍道 protein VP1 in baculovirus expre拍作ssion vector system（BEVS）has been 站近not reported till now工去. In the study，VP1 gen校做e was inserted into baculovirus v坐去ector of pFastBac I thr多自ough the restriction家男 enzyme site，the男東 identified recombinant 友區vector was transformed into DH10車公Bac，and the correct colo習內nies were screened by blue and w機風hite spots to extract the p腦麗lasmids that were transfected int雨影o Sf9 insect cells，an現雨d the recombinant baculovir問化us rBac-I-VP1 was店少 obtained after obvious cytopathic 事購changes. The expression o機不f VP1 protein was 商兵identified by Wester林對n Blot and indirect immunofluor從北escence（IFA）. The results sh著司owed that the titer of recombi多視nant baculovirus rBac-I-VP那冷1 was 6.8×105 pfu/mL；the哥土 expression products with relativ大東e molecular weight of about 27 k來雜Da could be detected by 冷電Western Blot，which could be reco體做gnized by SVA rabbit positive serum；市遠it was shown by IFA that，VP1 p們媽rotein could be expres科跳sed in Sf9 cells. 廠校It was concluded that th內鐘e SVA VP1 prote時東in expressed by the鐘到 BEVS was with good immunoreacti商南vity，which provid秒廠ed a basis for future study on re我術lated functions a兒國nd the development of 購能diagnostic reagents.全都報文下載鏈接：https://kn遠信s.cnki.net/kcms/detail/detail.aspx?dbco就場de=CJFD&dbname=CJFDAUTODAY&file司議name=ZGDW2020120時下22&v=Bh8a8ZT44%秒微25mmd2BVJojsL36nb煙兵%25mmd2BCImZ5SxS65hljHwT7w8vih那坐qJVTRfQYhRa61GRaSRLVI
2020-12-03
塞内卡病毒CH/ZZ/2016株全基因組測序與分廠子析
爲了解塞内卡病毒分離株SVA/CH/ZZ/2016月黃全基因組序列，設計4對(duì)相互重疊的特異性引物擴增關們基因片段，將(jiāng)擴增産物分别克隆至pCE2TA/Blunt-Ze也城ro載體并進(jìn)行測序，拼接校正後(hòu)獲得SVA/C話還H/ZZ/2016株全基因組。結果顯示，該毒株基因組全長朋校(cháng)7 292 bp，包括5'UTR（670 bp）話地、ORF（6 546 bp）以及3&雨司#39;UTR（76 bp）。選擇國(guó)内外其他小可9株參考毒株序列，對(duì)編碼區12個基因的核苷酸匠事及編碼氨基酸進(jìn)行比對(duì)。結果顯示，核苷酸同源性最高的是3B基農亮因，最低的是VP1基因，其餘基因的核苷酸序列同源性均在85.2%~10鐘可0%之間。VP1基因遺傳進(jìn)化分析顯示，SVA/CH/ZZ/2習西016株與美國(guó)分離株USAIL_Purd男藍ue_43_2016和USAIN_Purdue_3698_201音師6株親緣關系最近，屬同一進(jìn)化分支，與商女原始毒株SVV-001株親緣關系最遠。對章知(duì)SVA/CH/ZZ/2016株和原始毒株SVV-001 VP國房1蛋白的氨基酸序列進(jìn)行比對(duì)，發(f多窗ā)現共有10處氨基酸差異。本研究通過(guò)對(duì)SVA/C車地H/ZZ/2016株全基因組測序及著友分析，爲進(jìn)一步開(kāi)展錯機SVA分子生物學(xué)研究及流行病學(xué大家)調查提供了基礎數據。Sequenc花下ing and Analysis on Whole Genome of Sen森線ecavirus CH/ZZ/2016 St去筆rain In order to identify the whole ge學分nome sequence of se體樂necavirus SVA/CH/ZZ/20訊會16 strain，four pairs of overlappi水小ng specific primers were designe坐聽d to amplify gene fragm書頻ents. The amplified product嗎線s were cloned into pCE2TA/Blunt場南-Zero vector for sequencing. The w長視hole genome of SVA/CH/ZZ/2016 strai短時n was obtained after splicing 笑人and correction. The results showed tha電購t the total geno窗國me length of the strain was東海 7 292 bp，including 5'UTR（670 bp），O舊冷RF（6 546 bp）and 3'UTR（女畫76 bp）. Other 9 reference strains both看著 at home and abroad were selected 風地and compared with the nucleotide 外下and amino acid sequence身西s of 12 genes in cod呢說ing region. It was foun音醫d that the nucleotide homology was黑業 highest in 3B gene，minimum外去 in VP1 gene，and remained 85.2%兵技 to 100% in other genes. B計算y phylogenetic analysis，it was shown 她空that SVA/CH/ZZ/2016 strain had 道下the closest genetic re短商lationship with American iso校老lates USAIL_Purdue_43_2016 a好綠nd USAIN_Purdue_3698_2016，sh通少ared the same evolutionar街秒y branch，and was with larg機紅est evolutionary distance wit請是h the original strain SVV-001，雨對a total of 10 amino 討樂acid differences were found. Therefor區在e，the basic data was provided 老了for further study鐘聽 on SVA molecular biology and epi為嗎demiological investigation.全廠妹文下載鏈接：https://kns.cnki.net/kcms/deta也機il/detail.aspx?dbcode=資資CJFD&dbname=CJFDAUTODAY&filename=Z女科GDW202012021&v=Bh8a8ZT44%25mmd2BWe%25站你mmd2Fu6mMUO8Io9ewTzdK8XE6rL14BCn5ZgS見喝5VrStjQTvAjvZuRVCkn3
2020-12-03
QuEChERS法在獸藥殘留檢測中的優化與應用
獸藥的不合理使用可導緻動物産品金水及環境中獸藥殘留超标，從而對(duì)公共衛生安全構答計成(chéng)威脅。QuEChERS法是一類快速、簡便、經(jīn水劇g)濟、高效、耐用和安全的樣(yàng)話請品前處理方法，常用于植物産品農藥檢測。子能通過(guò)持續優化，目前該方法開(kāi)始在動物産品獸藥殘留檢測光場中得到應用。本文綜述了QuEChER大外S法的特點，探讨了該方法在畜禽産品或樣(yàng)品獸藥殘用兵留檢測中的優化與應用。與植物樣(yàng)品相比，動黑門物相關樣(yàng)品基質複雜，脂物市肪含量較高，因此需要對(duì)QuEChERS法進(j謝城ìn)行優化。常用的優化方式包括提取液優化、脫水鹽優化和吸附劑綠他優化。目前QuEChERS法在畜禽肌肉日物、内髒、雞蛋、牛奶及相關産品以拿會及尿液、糞便等環境樣(yàng)品獸藥殘留萃取中麗海得到應用。随著(zhe)與其他提取淨化話知方法的連用以及自動化提取裝置的不斷更新，未來QuEChERS法會(huì)得到紙大持續優化與發(fā)展，其提取效率和淨化效果將(jiāng)進(jì人熱n)一步得到提高，并將(jiāng)在動物産品殘留檢測中發(頻嗎fā)揮重要作用。Optimization an坐新d Application of Qu家雨EChERS in Detection o機門f Veterinary Drug Residues 媽呢The abuse of veteri開事nary drugs would lead to excessi也人ve veterinary drug residues i學綠n animal products an外鐘d environment，posing a thr音謝eat to public health. The 海公QuEChERS，which is quick，easy子有，cheap，effective，rugged and safe，i對如s a pretreatment對放 method for samples and usuall音低y used to detect pes話訊ticide in plant products. At 兒區present，the method with cont章微inuous optimiza紙玩tion has been applied in the detection 要河of veterinary drug會林 residues in animal samples. In the p飛城aper，the characteristics of the m不門ethod were summarized，it山哥s optimization and application in the 見算detection of veterinary drug 北志residues in livestock and poult白麗ry products or samples were discusse到體d. Animal samples were with comple但技x matrix and higher fat con們北tent compared to plant samples，so t工計he method should be optimized b刀一y common means includ機算ing the optimizations of extraction so南和lution，dehydration salt and ads小器orbent. At present，the QuEChER藍如S has been applied in the extraction o去站f veterinary drug re說雪sidues in animal mu鐵服scle，viscera，eggs，mil要答k and related product西朋s，as well as urine，feces and美可 other environmen鐵關tal samples. With c說聽ontinuous use of other extraction河拿 and update of automatic extraction習會 units，the QuEChERS would be continu醫他ously optimized and d麗雨eveloped in the future，and its extrac科年tion efficiency and purifica拿空tion effect would be fu作媽rther improved to play an家文 important role in det雨見ection of veterinary drug residues 妹算in animal products.全文下載鏈接：http公雨s://kns.cnki.net/kcms/detail/detai那了l.aspx?dbcode=CJFD&dbname=CJFDA術照UTODAY&filename=Z山自GDW202012020&v=Bh8a8ZT44%25mmd白件2BW2NuVVy8plweIK9k3ca%2近錯5mmd2B%25mmd2FPefx了歌CuLCIL0oso4h04Tfcw對他yV0b%25mmd2BfKoX13
2020-12-03
我國(guó)動物疫病風險評估研舞嗎究熱點和趨勢——基于CiteSpac資制e的可視化分析
爲探究我國(guó)動物疫病傳播風險評估的研究熱點及趨勢，以2000—2019靜歌年“中國(guó)知網”（CN用銀KI）數據庫收錄的348篇動物疫病風險評估相關文獻爲研究對(duì)象，懂訊利用CiteSpace軟件對(duì)文獻進(jìn)行物科時(shí)空分析和内容分析。時(shí)空分析顯示，這服會(zhè)20年間動物疫病風險評估文獻數量呈現從少到多再減車女少的動态過(guò)程，各大研究機構合作較少。内容分析顯示，研究熱見車點主要涉及“風險評估”“風險分析”和“動物疫病防控”等方內車面(miàn)。提示研究機構應加大動物疫病風險評估研究力度，加強各大紙訊研究機構間的合作，推進(jìn)動物疫病風險評估的研究、錢風分析、監管，完善預警機制，進(jìn)一步提升我國(guó)動物疫病防控水平雜事。本研究可爲深化動物疫病風險評估研究、實踐和預警提快書供參考。Research Hotspots and Trends of 視美Animal Disease Risk Assessment in 土黑China According to Visual Anal空快ysis Based on CitespaceIn order 線慢to explore the research 近老hotspots and trends of animal d外師isease risk assessment in China內雪，348 articles concerning such risk a線章ssessment registered in the database of是區 CNKI during 2000 to 20可要19 were retrieved and音笑 analyzed by CiteSpace 化老software based on their time and space不山 as well as contents到電. It was shown that，by 雨多the temporal and spatial analy雪個sis，the number of li為費terature on risk assessment went throug和議h a dynamic process中一 from emerging to increasi多窗ng then to decreasing 外愛in the past 20 years，and f錢遠ewer major institutes c作吃ooperated with 海算each others. The contents of the litera女離ture were analyzed，it 民影was found that“risk 數新assessment”，“risk analysis”and“也事animal disease preventi外少on and control”w信技ere the research hotspots.年議 Therefore，institutes shou大看ld strengthen their inputs in animal的件 disease risk assessment，and incr鐵煙ease cooperation with each others，so鄉明 as to push forward the stu自鐵dy，analysis and supervision 低門concerning risk習拿 assessment，improve early warning件音 system，and to further upgrade 影訊the control level of animal dise通多ases. Some references綠鐵 were provided to f明綠urther the study and practice of risk 錢就assessment and early warni那紙ng.全文下載鏈接：https哥城://kns.cnki.net/kcms/detail/detail.a短為spx?dbcode=CJFD&d民討bname=CJFDAUTODAY&器舞filename=ZGDW202012019&v=Bh8a8湖我ZT44%25mmd2BUmenw8z1%25mmd2習國BeFZoLuHtJ0BefrEsn2GG6E6t0W中作HpgBiQ2tAHubGLfwaZt
2020-12-03
我國(guó)非洲豬瘟相關技術專利發(fā)展現狀
爲了解我國(guó)非洲豬瘟（ASF）相司民關技術專利研發(fā)重點，利用中國(guó)專利文摘數據庫（CN老票ABS）等專利數據庫，結合“人工閱讀去噪”篩選出295件ASF相關技術專利麗很，并運用Excel軟件對(duì)2006年1月至2湖弟020年7月專利的申請量、技術領域、申高通請人、研究核心等進(jìn)行了分析。結果顯示：我國(guó)專利總體申請可關量呈現增長(cháng)趨勢，并從2018年開(kāi)員用始出現爆發(fā)性增長(chá林個ng)；ASF技術專利以快速、高效、特異地鑒農鄉定和監測ASF病毒以及有效、安全地預防和治療ASF爲研究的重點和熱點；專場農利申請人涉及面(miàn)廣，黃校涵蓋企業、科研院所、高校和個人，其中企業申劇門請總量最高，其次是科研院所和高校；研究核心各有側重，科研院所和高校事錯側重于基礎技術的探索和研發(fā)，而企業則偏重于技術和産業化的有效結合外電。随著(zhe)我國(guó)ASF疫情防控重要性和緊行做迫性的日趨增加以及研究手段的日益更新，可以預期科算錢研院所、高校以及企業等各申請人在該領域中的研究將(j快區iāng)會(huì)更加活躍。本文爲我高明國(guó)ASF防控及其技術研發(fā)等提供了參考。行光The Development of Technical Pate器一nts Related to Afric自科an Swine Fever in Chi去子na In order to identify the res大雪earch priority of technical paten要文ts related to Africa swine fever（ASF間房），295 relevant patent慢身s were selected by use of patent d吃要atabases including China Patent Ab山書stracts Databas高大e（CNABS）and in combination with“黑農manual reading denoising”，and the資從 number of application，technical f看文ield，applicant and re我還search core of patents from January 200會如6 to July 2020 were a湖女nalyzed by the Exce少銀l software. The results showed t她舊hat the total number of 車木application was increasing and had boom年科ed since 2018；the tec物看hnical patents fo吧行cused on rapid，effective and spe做音cific identification an在著d detection of African swine 暗鐵fever virus（ASFV）and safe prev風務ention and control of ASF；the app民家licants might come from enterprises，i弟人nstitutes，colleges or be an indivi作錢dual，most of them were from enterpr生月ises，followed by institut哥訊es and colleges；for the researc北自h core，institutes 電化and colleges focused輛湖 on the development of 多他basic technology影他，and enterprises mainly光業 developed the effect我場ive combination of technology and indu上市strialization. With 水北the increasing importance and urg得劇ency of ASF con錯聽trol as well as the update of researc窗子h methods in China，it男吃 could be expected that the 秒場research by the applicants would be mo輛制re active in the 湖到field. Some refere你空nces were provided for prevention and 厭路control of ASF as we黃樂ll as the development of releva從嗎nt technologies in China.全文下載鏈接：h南間ttps://kns.cnki.n跳錯et/kcms/detail/信些detail.aspx?dbcode=CJF在外D&dbname=CJFDAUTODAY和很&filename=ZGDW202012018&v=Bh8a8ZT44%25m吧兒md2BWtVTc8gWyxxX6VayE8N有空ViQNoQb6N4Khk0FzS開去xwn7eXtxiG3PSxi員光VOt
2020-12-03
非洲豬瘟新型疫苗研究進(jìn)展
非洲豬瘟（African swine fever，看麗ASF）已存在約一個世紀，給世界養豬業帶來巨大損歌鐘失和威脅。爲此國(guó)内外科學(xué店微)家一直緻力于ASF疫苗研究，從早期在綠的滅活疫苗、弱毒活疫苗到目前的新型疫苗，包括基因缺失減毒活疫苗、對商亞單位疫苗、病毒活載體疫苗、DNA疫苗、單周期和熱病毒疫苗等。但直到現在，世界上仍未制造出切實有效并投入生産的ASF疫苗線化。研究發(fā)現：滅活疫苗不具有保護性或僅具有部分保護性；亞單位疫數飛苗具有部分保護性；基因缺失減毒活疫苗被(bèi)證實具有同源保護性，但其安全姐東問題仍未得到解決；病毒活載體疫苗雖會(hu站小ì)産生特異性抗體，但仍需要進(jì跳男n)一步研究；DNA疫苗雖然能(néng)夠誘導産生體液免疫和細胞免疫，但不能懂農(néng)提供保護。目前，一種(zhǒng)新型的單周期病毒疫分場苗正被(bèi)研究，也爲ASF新型疫苗研制提供了新選擇。随著(湖音zhe)基因工程技術與分子生物學(xué)技術的不斷發(f校鐘ā)展以及對(duì)ASF病毒身哥基因組研究的不斷深入，ASF疫計視苗有望研制成(chéng)功。Research錯能 Progress of Novel Vaccines間鐵 against African Swine Fever Viru能一sAfrican swine feve資哥r virus（ASFV）has北歌 remained for ab和報out a century，which has brought huge e開吃conomic losses and threats銀那 to the world. Therefore，the 小一study on vaccines against ASFV has店土 been continued 習上by scientists both at home and abroa風什d，from the earl朋謝y inactivated vacci草術nes，attenuated live vaccines to cur美火rent new vaccines，inclu那習ding gene deletion at自舊tenuated live vaccines，subunit vacc又這ines，live vector vaccines地哥，DNA vaccines，single-cycle virus vaccin窗能es，etc. But up to 不快now，no effective vaccine had been de又風veloped or produced. It w現行as studied that inactivat鐘跳ed vaccines were not protective or還見 partially protective；subunit金老 vaccines were partially protective文話；gene deletion live attenuated vaccin綠都es had been proven to be homol文音ogous-protective，but th器村eir safety had not been solve紙門d；although live vector vaccines 子愛could produce specific antibodies廠錢，further research was still needed；DN窗土A vaccines could兒白 induce humoral or cell子書ular immunity，but failed放務 to provide protecti信間on. At present，a kind of novel sin舞熱gle-cycle virus vaccine was being習讀 studied，which also provided a 在個new option for the de多吧velopment of novel vaccines agains月河t ASFV. With the continuous dev電會elopment of genetic答水 engineering technology a東湖nd molecular biology technology as w跳舞ell as the in-depth study on ASFV 吃科genome，vaccines against ASF were少電 expected to be successfully develop車工ed.全文下載鏈接：https://kns.cnki.ne報黑t/kcms/detail/detail.aspx?dbc線都ode=CJFD&dbname=CJFDAUTODAY服愛&filename=ZGDW202012017&v=Bh8a8ZT44%25m裡吃md2BXHiNGRu1LlBDMWBctkn9umG3qE湖廠8vPysHtqUmRpYD0vEXOEXJv76oGP制玩
2020-12-03
轉基因動物及其産品檢測技術研究進(jìn)展
爲探索可靠、靈敏的轉基因動物及其産品檢測技術，從會道核酸檢測和蛋白檢測兩(liǎng)個方面(miàn)，綜述了轉基因動物及其産來刀品不同檢測技術的原理、特點及應用，提學理出了每種(zhǒng)檢測技術的不足及今後(hòu)銀從轉基因檢測技術的發(fā)展趨勢和研草遠究方向(xiàng)。目前，已經(jīng)研發(fā)的核酸檢測技術主要不裡包括多種(zhǒng)PCR方法、環介導等溫擴增技術、South呢吃ern blot技術和染色體原位紅電雜交技術等，蛋白檢測方法主要包括酶聯免疫吸附檢測技術、免疫層析試他行紙條、Western blot技術、免疫匠業組織化學(xué)和免疫熒光技術等。三代測序技藍明術是近年來提出的一種(zhǒng)新型基因檢測技術，适用于對森得(duì)未知序列轉基因動物的分析服吧。随著(zhe)轉基因技術的發(fā)展，轉話睡基因動物及其産品越來越多，高通量、高靈敏綠的度和低成(chéng)本的檢測技術將(jiāng)成(chén哥金g)爲未來轉基因動物及其産品檢測的研究重點。 Researc長離h Progress on the Techniqu裡電es for Testing Transgenic睡筆 Animals and Th線跳eir ProductsIn order to explore a船明 kind of reliable and sen有門sitive technique for test嗎亮ing transgenic anima風訊ls and their products，the pri少公nciple，characteristics and app多信lication of relevant techniques were su長秒mmarized from the aspects of冷藍 nucleic acid and protein det的資ection，then the disadvantages 校站of each technique as well as relevant照樂 development trend and direction樂公 to study were put forward. At pres服頻ent，the established tec務文hniques for detec錢為ting nucleic acids mainly的笑 included PCR，rin高那g mediated isothermal amplificatio中也n，Southern blot and chromosome in 小是situ hybridization，and those for prot店西ein included enzyme-linked 工書immunosorbent assay（ELISA），聽和immunochromatographic strip，W道高estern blot，imm服鐘unohistochemistry and immunofluor就多escence assay. The th器厭ird-generation sequenci厭愛ng technique had been used to detec路如t gene sequences in recent ye銀鐘ars，which is appropriate t錯上o analyze transgenic anim大城als with unknown seq店女uence. With the development of transg來要enic techniques，the quantit讀吧y of transgenic animals 遠暗and their products was inc這森reasing，so the techniq們關ues with high throu理畫ghput，high sensitivi街物ty and low cost will become a priority但訊 in the future. 全文下載鏈接：https://k議報ns.cnki.net/kcms/detail/d裡到etail.aspx?dbcode妹關=CJFD&dbname=CJFDAUTODAY是對&filename=ZGDW2放算02012016&v=Bh8a頻農8ZT44%25mmd2BVv3mciFa8kv4銀長rOzUDjKYUNajRapQZ5zhlukwGnapMO音山diSX0e2zkBbw
2020-12-03
口蹄疫通過(guò)雲南中緬邊境活牛走私傳入我西報國(guó)的定量風險評估
受國(guó)内市場需求和價格等影響，在筆會與緬甸、老撾接壤的雲南邊境地區，活牛走私現象屢禁不止，成(chéng)爲口蹄疫分化傳入我國(guó)的主要風險途爸秒徑。爲分析雲南中緬邊境活牛走私傳入什美口蹄疫的可能(néng)性，收集雲南邊境地區活牛非法調入途徑車行、路線和數量等信息，利用“情景樹”法建立傳入風險随機模型進(jìn)行仿真分笑海析。結果顯示：從雲南中緬邊境走私調入1頭牛，口蹄下下疫傳入我國(guó)的概率爲0.81%（95%CI，0.43%~1.嗎個47%），且每年約有14 017頭在做（95%CI，7 520~25 660頭）口蹄疫感染牛自中歌書緬邊境走私進(jìn)入雲南省；口蹄疫通過(guò)中緬東路邊境活牛走私傳入我國(guó)的風險取決于緬甸北部活牛市場要什内的口蹄疫流行率。結果表明，雲南中緬自作邊境活牛走私導緻口蹄疫傳入我國(guó)的可能(néng)做拍性不容忽視。建議對(duì)緬甸活牛以及過(guò)境緬甸活牛入境前進(jì她喝n)行口蹄疫疫苗免疫，并在雲南當地有效監管下屠宰；允許從做謝東南亞國(guó)家進(jìn)口符合我機制國(guó)進(jìn)口檢驗标準的冷凍去骨肉。本研究首次多還對(duì)口蹄疫通過(guò)中緬邊境活牛走私傳入我國(guó)的風險進(j女器ìn)行了定量評估，爲我國(guó)口蹄疫防控提供了重要信息和數據。Qua到黑ntitative Risk Assessment on FMD錢小 Introduced into Ch媽秒ina via Smuggled Live Cattle across Chi長哥na-Myanmar Border in Yunnan Provin高新ceAs being affected by the need a服下nd price in domestic markets，activ兒物ities of smuggl亮在ing live cattle have continued in Y市又unnan frontier areas b北鐘ordering Myanmar and Laos in spite離會 of repeated prohibition，which have po習銀sed an important risk 道理pathway for foot and 購國mouth disease（FMD）to林吃 be introduced into Chin請道a. In order to analyze the probabili都中ty of FMD introduced into Ch下開ina via smuggled liv低樹e cattle across China-Myanmar b道站order in Yunnan P弟理rovince，collect the informa些男tion concerning t東裡he pathways，rout農答es and quantity of illegally im們這ported live cattle，a stoch分坐astic model of ent區吃ry risk was establis朋議hed and analyzed by the“scene tree”. Th文事e results showed河是 that when one cattle was smuggled劇愛 via the border，the probabi厭刀lity to introduce F喝時MD into China was 0.81%（95體理%CI，0.43%–1.47%），what 高上was worse，about 14得黃 017 cattle（95%CI，720–25 660 黑長cattle）infected with FM民你D were smuggled in得區to Yunnan Province every y男船ear；and such risk 資銀depended on the prev土多alence in cattle markets in northern 厭時Myanmar. It was co通長ncluded that the probability to黃議 introduce FMD into Chin道金a through smuggling live cattle f低生rom Myanmar should not be ignor木坐ed. In view of the abo農銀ve，the live cattle that were市玩 from or transit via Myanm好問ar should be vaccinated a他玩gainst FMD prior to entry，雨門and slaughtered under effect線黑ive supervision by loca人拍l authorities；a計分nd frozen boneless meat公愛 that conformed to China's imp黃科ort inspection standards sho腦妹uld be allowed to城飛 be imported from Southeast Asia身哥n countries. In t計美he study，the risk to introduce FMD 鄉山into China via China-Myanmar border 村哥was quantitatively些匠 evaluated for the first time，司遠which provided important inf通公ormation and data for 笑了the prevention and co黃知ntrol of FMD in China.全文下載鏈接：h低兒ttps://kns.cnki.net/kcms/detail/d樹煙etail.aspx?dbcode=CJFD&dbname=CJ短對FDAUTODAY&filename=ZGDW202012002&v=Bh8我很a8ZT44%25mmd2BUG3I6hkBCxo4jms8VPlghmnI笑內EoyLK0k1b1hIhtiJ70eTzdWrw腦東J8QAs
2020-12-03
毛皮動物新冠病毒監測簡報
2019新型冠狀病毒病（COVID-19）給公共衛生安全帶來巨大危害，但目前村雪引起(qǐ)該病的新型冠狀病毒（SARS-CoV-亮街2）來源還(hái)未有定論。森謝現有證據顯示，SARS-CoV-2與分離兒白自蝙蝠和穿山甲的 SARS-CoV-2病毒株同源性相近信樹，表明蝙蝠和穿山甲可能(néng)作爲SARS-CoV-2影冷的中間宿主存在。目前，丹麥、荷蘭和美國(guó)等國(guó區是)家已有人工飼養水貂及飼養場工作人員感染SARS-CoV-2的報道(d會站ào)。試驗研究顯示，SARS-CoV-2可在雪就遠貂上呼吸道(dào)複制，且雪貂對(duì)但化SARS-CoV-2高度易感，表明雪貂可作爲一種(zhǒng)SA長著RS-CoV-2感染和傳播的動物模型。2020年2—7月家廠，本課題組對(duì)山東、吉林、遼甯、黑龍江等毛皮動物主要養她公殖省份的水貂樣(yàng)品進(jìn)行SARS-C見票oV-2病原及抗體監測。利用WHO推薦的real-time R黑熱T-PCR檢測方法，對(duì)328份水貂内髒組織及咽拭子樣(y頻自àng)品進(jìn)行檢測，結果所短時有樣(yàng)品均爲SARS-CoV-2麗習陰性；采用北京萬泰生物藥業有限公司提供醫下的SARS-CoV-2抗體檢測EL站窗ISA試劑盒，對(duì)35份水貂血清樣(yàng)品進河那(jìn)行抗體檢測，結果樣(yàng)品中抗SAR呢國S-CoV-2抗體均爲陰性。由于國(guó)内養殖多爲密集型庭院養殖，養殖人唱拿員與水貂密切接觸，且國(guó)外已有養殖場水貂答費大規模感染SARS-CoV-2的先例，爲保障毛皮動物行業健康發(fā)業還展，此次監測重點對(duì)我國(guó)北方地區毛皮動物養殖場進(jìn行兵)行采樣(yàng)，結果并未發(fā)現毛皮動物攜帶SARS科校-CoV-2和針對(duì)SA訊照RS-CoV-2的抗體。這(zhè)可能(né女拿ng)與目前我國(guó)人群的低SARS-CoV-2感染率和未有養殖什綠場工人感染有關。本實驗室已建立了檢測毛皮動物及寵物的SARS-Co麗物V-2抗原及抗體檢測方法，在目前全球COVID-19疫情的嚴峻形勢下，未船船來仍需對(duì)毛皮動物及寵物進(jìn)行長(cháng)期的SARS-新問CoV-2流行病學(xué)監測。Overview on th飛那e Surveillance of SARS-CoV-光船2 in Fur Animals The coronav你請irus disease 2019（COVID-19文不）has brought great ha器農rm to public health safety，but th那裡e source of severe acute 銀資respiratory syndrome 又理coronavirus 2（SARS-CoV-2）causing COVID近書-19 has not been concluded. I草鄉t was shown that，by available evide草爸nces，the virus was with similar 坐風homology with the SARS-CoV-2 iso輛森lated from bats an高吧d pangolins，whic時影h indicated that bats and pan呢醫golins might act as the intermedia影頻te hosts for the virus.秒照 It was reported tha商化t captive minks and farm workers had b麗影een infected with SARS-CoV-2 北明in Denmark，Netherlan靜討ds and the United States. According to 綠請related study，SARS-CoV-2 could 鐘吃replicate on the up討水per respiratory 笑和tract of ferrets who were 生雪highly susceptible t分讀o the virus，which indicated that f能朋errets could be used as an animal mode南科l of infection an工爸d transmission of the路放 virus.In February to July懂湖 of 2020，the mink samples collected空吃 from Shandong，Jilin，Liaoning，He銀月ilongjiang and other provin新現ces where most fur animals were k謝小ept were monitored for pathogen and ant你校ibody of SARS-CoV-2，328 visceral tissu什學e samples and throat swabs were朋行 detected by the real-time RT-PCR as志這say recommended by WHO，and all the 亮我samples turned out to be negative；mean到海while，35 serum samples were麗樹 detected by us就國e of the ELISA kit p明機rovided by Beijing Wantai Biologi外跳cal Pharmacy，and all小她 the samples were negativ影司e for the antibodies against SARS-水行CoV-2. The operators had been i飛人n close contact with minks d兵理ue to intensive courtyard farming，es道輛pecially，there had been cases of mi林聽nks infected with SARS-CoV習科-2 on a large scale in foreign count廠多ries. In order to safegua紅電rd healthy development of f但行ur animal industry，the farms were mo看讀nitored and sampled as a priority森朋. No SARS-CoV-2 or relevant antibody w厭理ere found in any fur animals，懂好which might benefit from low infection能見 rate in population事可s and the absence 村花of infection in關報 farm workers in China. The meth林訊od for detecting pathogen a市快nd antibody of SARS-Co行答V-2 in fur animals a見是nd pets was established by our la音那boratory，and epid大如emiological surveillance for SAR得師S-CoV-2 in fur animals and pets shou城腦ld be continued for我師 long term in the fut視吧ure in response to current弟懂 severe situation of COVID-19 across章讀 the world.全文下載鏈接：http拍哥s://kns.cnki.net/kcms/detail/detail.asp外計x?dbcode=CJFD&dbname=CJFDAUTODAY&fi機服lename=ZGDW202012001&v=你物Bh8a8ZT44%25mmd2B子自V6mndOdWwC3lCFYz%25mmd體的2F9tG%25mmd2B0Yu7LUoQYiF8eJU%2飛科5mmd2FuGJY16mnuTkcHAh0e
2020-12-03
重組禽流感病毒（H5+H7）三價滅務著活疫苗免疫雞群的臨床效果評價
爲評價重組禽流感病毒（H5+H7）三價滅活疫苗（H5N1 Re-11株+Re個服-12株，H7N9 H7 Re-2株）免疫雞群的臨床應東著用效果及當前禽流感免疫程序的合理性，對(duì)河南省使用重組房女禽流感病毒（H5+H7）三價滅活疫苗免疫的940個存欄500好科隻以上的養雞場開(kāi)展臨床應用效果調查，并抽取328個場鐘我開(kāi)展免疫抗體檢測。結果顯示：免疫後(hòu)1周内出現讀工不良反應的養雞場190個，占20.2醫水1%（190/940）；發(fā)病的養雞場140個，占14.89%（140/業裡940）；出現死亡的養雞場51個，占5.43%（51/940）。免疫後(hòu文文)1周内出現不良反應的雞隻占0.11%，快醫發(fā)病的占0.09%，死亡的占0.04%。血清樣(yàn黑下g)品的血凝（HA）和血凝抑制（HI）試驗結果顯示：H5亞型Re-11株錯大場群合格率爲97.56%，免疫抗體陽性率爲94.20%；H5亞型Re-12株場多空群合格率爲96.34%，免疫抗體陽性率能校爲92.70%；H7亞型Re-2株場群合格率爲98.17話路%，免疫抗體陽性率爲94.67%。結果表明船歌：重組禽流感病毒（H5+H7）三價滅活疫苗的雞群暗內免疫效果較好(hǎo)，免疫後(hòu)的臨床雨村不良反應率極低。但在免疫過(guò)程中，應盡量消除影廠靜響疫苗臨床效果的不利因素，定期開(kāi秒來)展免疫效果監測，及時(shí)調整免疫程序。本研究爲全國(guó)區下高緻病性禽流感疫苗的應用提供了參考。Evalu船自ation on the Clinical Ap飛用plication Effect of Rec計作ombinant Avian Infl不件uenza Virus（H5+H7）熱頻Trivalent Inact化睡ivated Vaccines in Chicken東空s In order to evaluate the clin書就ical application effec生草t of recombinant avia購說n influenza vir身知us（H5+H7）trivalent inactivated vaccines務購（H5N1 Re-11+Re-12 strain and電影 H7N9 H7 Re-2 strain）in chicken鐘算s and the rationality of c雪一urrent vaccination pr時媽ogram，940 farms with鄉服 more than 500 chicke數亮ns were investigated in Henan 件樂Province，in which，328 fa員朋rms were selected 章錯for detection of immune antibody訊了. It was found that，within one煙信 week after vaccination，some ad南都verse reactions reve高兒aled in 190 farms，acco笑信unting for 20.21%（190/940）；知作avian influenza outbreaks照亮 in 140 farms were detected，很紙accounting for 14.89%（140/9生理40）；and dead cases 厭公were found in 51 farms，accounti們不ng for 5.43%（51/940）. Th公做e proportions of chickens wi就鄉th adverse reactions，bei嗎紅ng infected and dead were 0.11%，0.09% 章慢and 0.04%，respectively. It was fo他內und that，by hemagglutinat地志ion（HA）and hemagglutinati河作on inhibition（HI）test f學器or serum samples，the farm/fl醫歌ock qualified rates of H5 Re-1腦數1，H5 Re-12 and H嗎通7 Re-2 strain were 97.56%，96.34% and但熱 98.17%，respectivel唱熱y，and their positive rates of immun著門e antibodies were 94.20%，92.70% and理匠 94.67%，respectively. It wa海照s concluded that 購外the recombinant avian influenza v物個irus（H5+H7）trivalent inactivated vacci紙空ne could receive good effect 自木in chickens，with lower le還這vel of adverse reaction.地美 However，any adverse 電日factors therefro務城m should be eliminated to the 舊技greatest extent during the 們來process of vaccination，which shoul鐘身d be monitored regularly，and the 秒畫vaccination prog樹他ram should also be timely a錯東djusted. Some references were provi快船ded for the application of vaccines a少討gainst highly pathogenic a票計vian influenza vir問得us in China.全文下載鏈接：https://kns.中嗎cnki.net/kcms/detail/de友藍tail.aspx?dbcode=你美CJFD&dbname=CJF高吧DAUTO&filename=ZGD會舞W202011022&v=Bh8a8ZT44%樹路25mmd2BViTsUIpnMUhMd國場fl8JdNVN4nOcJGcWypRTOzA9syj9問劇XPAWc4KBTGMQp
2020-11-06
高緻病性豬繁殖與呼吸綜合征病毒自然感染與人身子工感染下的組織病理學(xué)觀村近察
爲進(jìn)一步研究高緻病性豬繁殖與呼吸綜合征病毒（章呢HP-PRRSV）的緻病機理，對(du很習ì)HP-PRRSV自然感染和人工感染下的病理組織學(xué)變山朋化進(jìn)行比較觀察。在組織病理學(xué風費)變化方面(miàn)，自然感染病例病變性質與人工感染基本相同，但在病變範圍歌計以及嚴重程度方面(miàn)存在銀友明顯差異：自然感染病例的胃腸道(dà但那o)病變尤爲顯著，而人工感染病例卻較少引發(fā)消化道(dào)病變；自然友個感染病例的炎症波及全身淋巴結，而人工感染病例以肺門淋巴結、下颌淋巴結、腸系睡森膜淋巴結病變最爲明顯，其他部位淋巴結未見明顯病湖拍變。自然感染病例中，常見多種(zhǒng)病原的混合遠要感染，提示HP-PRRSV可能(néng就開)作爲多種(zhǒng)疾病的基礎病因。自然感染和人工感染病例在肺髒信快上均表現爲嚴重的間質性肺炎，說(shuō)明肺們時髒部位的病理變化對(duì)單純HP-PRRSV感染的現拍診斷具有指證意義。本研究爲HP-PRRSV感染的診斷及其緻病機理研究提照從供了理論依據。Histopathological Obse睡刀rvation on Natural and Artificial Infec個喝tion with HP-PRRSVIn o林不rder to further study the pathog微如enesis of highly pathogenic porcine時間 reproductive and respiratory syndrome 低化virus（HP-PRRSV），the histopathol河煙ogical changes o體車f natural and artificial in費上fection with HP-PR頻道RSV were compared and o月微bserved. For the nature of公森 lesions，the natural and art男她ificial infection cases w船雜ere basically similar，but obvio長答usly different in th那厭e scope and severity degree of t木術he lesions；the gastrointestinal l訊水esions were particularly significa民鐘nt in the natural infection cases，and 年北fewer gastrointestinal 厭影lesions were obs時信erved in the artif事樂icial ones；all the ly朋上mph nodes were involved i文風n the inflammati山西on of the natur討司al infection cases，b頻吃ut the lesions of hilar lymp藍窗h nodes，mandibular lymph nodes and mes畫微enteric lymph nodes appe來資ared in the artificial 知工infection cases，and other lymph nodes w老黃ere normal. Mixed infection with 長業multiple diseases were common熱這 in natural infection cases，indicating水大 that HP-PRRSV might be the 火用general cause for several diseases從訊. Severe interstitial pneumonia音化 was shown in the l門關ungs of both natural 小金and artificial infectio大讀n cases，which indicated在相 that the patho友風logical changes of the lungs體務 were indicative fo間資r the diagnosis of single infection 老玩with HP-PRRSV. 熱自As a conclusion，a 裡家theoretical basis was provided for t家窗he diagnosis of 個站HP-PRRSV infection and for the study o南店n its pathogenesis.全文下載鏈接：h務明ttps://kns.cnki.net/藍問kcms/detail/detail.aspx?dbcode=C拿冷JFD&dbname=CJFDAUTO&fi玩開lename=ZGDW202011021&v=Bh8a8ZT44%25mmd呢女2BUVIUjuVi2E9SXrkHJjA%25mmd2F8oe他窗zzoQP1ttRyl97yDc5jZ6uBPwCc0西個mNOi
2020-11-06
重組PCV3 Cap蛋白在酵母細胞中的表達與鑒雪男定
豬圓環病毒3型（PCV3）是影響我國(guó廠水)養豬業的重要病原之一，目前尚無商品化疫苗用于防控該病。Cap蛋白藍費是PCV3主要的抗原蛋白。爲構建高水平表達可溶性計器PCV3 Cap蛋白的工程菌株，對(duì)Cap蛋白N匠愛LS序列進(jìn)行定點突變說村，構建重組表達載體pPICZαA-PCV3，經(j對她īng)轉染、酵母細胞表達以及純車校化，最終獲得可溶性PCV3 Cap蛋白。随後(hòu少內)對(duì)純化蛋白進(jìn)行錢文Western Blot驗證及電鏡東照觀察。結果顯示：獲得的PCV Cap蛋知有白大小約27 kDa；電鏡下觀察可見均一的病毒樣(yàng)顆粒（VLP去綠），顆粒直徑約18 nm。PCV3 Cap冷間蛋白在酵母細胞中的成(chéng)功表達爲進(jìn)一步開(kāi)路高發(fā)Cap蛋白亞單位疫苗雪金奠定了基礎。Expression and Identification of腦大 Recombinant PCV3 Cap Protein in Yea小術st CellsPorcine用綠 circovirus type 3（P費裡CV3）is one of the most serious patho了雪gens that would affect pig i鐵兒ndustry in China，but no 視可commercial vaccine has been available學但 till now. In order to e信文stablish an engineered strain 舞山that could express solubl拍現e PCV3 Cap protein at a high level，現唱considering that Cap pr現街otein was the main antigenic prot一吃ein of PCV3. The nuclear loc近空alization sequence（NLS）of Cap protein w如愛as specifically mutated to 文為establish the recombinant plasmid pPICZ金飛αA-PCV3，after t中司ransfection of 男子the plasmid as we森低ll as expression a服市nd purification of reco資師mbinant protein，the 鄉站soluble PCV3 Cap pr金線otein was obtained. Then the pu樹門rified protein was ver如公ified by Western Blot音白 and observed unde器風r electron microscope. I哥新t was concluded that the PCV C書紅ap protein obtained was about 靜河27 kDa，and unifo日讀rm virus-like particl空什es（VLP）with the diameter of 18聽知 nm were observed u高愛nder electron microscope. The 市黃successful expression of PCV3 Cap 化遠protein in yeast cell兒外s laid a foundation for further devel子師opment of Cap protein subuni草務t vaccines.全文下載鏈接：h樂電ttps://kns.cnki.net/kcm慢作s/detail/detail.aspx?dbcod科船e=CJFD&dbname=CJFDAUTO&filename志哥=ZGDW202011019&v=Bh8a8ZT劇讀44%25mmd2BW%25mmd2B2F5ndRK1P%25mmd2FQz車舊KJxnpMmCaZMw9aPlw6lKWcwzPB1vWxk9I2B高錢Rd7GJ
2020-11-06
同源血漿及血清中藍舌病病毒抗體ELISA檢測結果比較
爲簡化動物血液樣(yàng)品中藍舌病病毒（BTV）檢測及研上費究方法，采用2種(zhǒng)酶聯免疫吸附試驗（EL學少ISA），對(duì)采自雲南省德宏州芒市一家肉牛養會金殖場的30對(duì)牛同源血漿及血票會清樣(yàng)品進(jìn)行BTV讀為總抗體及VP7抗體檢測。BTV總抗體ELISA檢測結果顯示，同源血漿與血清之間議喝的陽/陰性結果一緻率爲100%，ΔPI在±5%以内的樣(yàng)品銀間占87%；VP7抗體ELISA檢測結果顯示，都校同源血漿與血清之間的陽/陰性結果一緻率爲93%，Δ草在OD在±0.05範圍内的樣(yàng)品占87%。結果表明：利用E好雪LISA檢測動物血樣(yàng)中BTV抗體時(shí)，血漿樣(yà話還ng)品可替代血清樣(yàng)品；血漿畫要樣(yàng)品的溶血程度不影響檢測結作朋果，但粗制血漿中的非溶解性雜質可能(néng)影響檢測結校書果，需要使用離心除雜後(hòu)的血文些漿樣(yàng)品。Comparis好又on of the Test Results o放件f Antibody against Bluetongue Vi化化rus in Homologous Plasma and Serum by 書媽ELISAIn order to simplify the 鐘線test and research meth海場od of bluetongue virus（BTV）術河in animal blood samples，30 pairs of bo唱一vine homologous p哥影lasma and serum sample習窗s collected from a beef厭樂 cattle farm in聽鄉 Mang City，Dehong Prefecture，Yunnan Pro視船vince were tested for antib用暗odies against BT通師V and VP7 protein. The results冷報 showed that，for the舞票 detection of total anti街村bodies of BTV，the consiste鄉風ncy rate of the positive/negative resul微資ts of homologous plasma and 商南serum was 100%，and samples with ΔP開些I less than ±5% accounted for 87%；as t空喝ested by VP7 Ab水如 ELISA kit，the 吧習consistency rate of the positive/道件negative results of homologous pla明線sma and serum was 照吧93%，and samples with ΔOD le吃那ss than ±5% accounted for 87%. 林在Therefore，serum samples m自月ight be replaced by plasma samples wh廠湖en the antibody against BTV為姐 in animal blood wa匠筆s tested by ELISA，an物答d the test resu什間lts could not be affected by the化司 hemolysis level o笑還f plasma samples rather than the in日數soluble impurities in crude她火 plasma which should be removed 土著by centrifugation before being 藍司used.全文下載鏈接：https://kn影鐘s.cnki.net/kcms/detail/deta物就il.aspx?dbcode=老都CJFD&dbname=CJFDAUTO但多&filename=ZGDW20朋亮2011018&v=Bh8a8ZT44%25mmd2BX2Lfl3ua白短iMzu%25mmd2Bl%25mmd2B1qun3qkzoQgjV玩放sMP8i3HGEkCPfr%25mmd2Fs月綠dtRDXv8w98
2020-11-06
單重PCR鑒别犬4種(zhǒng)布魯氏菌檢測方法的建立
爲建立适合犬布魯氏菌病臨床檢測的單重PCR方法，通過(guò)對(duì)比粗南少糙型（犬種(zhǒng)）與光滑型（羊種(zhǒng)、牛種(zhǒng)麗舊和豬種(zhǒng)）布魯氏菌基因組DNA的序列差異，設計湖為1對(duì)引物并優化反應條件，建立了可初步鑒别犬4種(zh些腦ǒng)布魯氏菌的PCR方法，然後(hòu)對(duì)該方法的音來特異性和靈敏度進(jìn)行評價，并對(duì)20份犬布魯氏菌血清學(x離木ué)陽性的血液樣(yàng)品進(jìn)行臨床檢測請懂。結果顯示，利用建立的PCR反應體系弟體對(duì)牛種(zhǒng)、羊種(z答水hǒng)和豬種(zhǒng)布魯氏菌均能(néng)爸鐘擴增出717 bp的目的條帶，對(duì)犬種(zhǒ用習ng)布魯氏菌能(néng)擴增出366 bp的目的條帶；最低知友可檢測到犬種(zhǒng)、羊種(zhǒng)、豬種說術(zhǒng)和牛種(zhǒng)布魯呢紅氏菌基因組DNA濃度分别爲5.07×10-2新哥、6.20×10-2、7.80×10-3和5.站也50×10-2 ng/μL；20份血液樣去民(yàng)品中共檢測到3份犬種道離(zhǒng)布魯氏菌陽性樣(yà對可ng)品，與使用GB/T 1864話街6—2018引物的PCR檢測結果一緻。結果表雨黑明，本試驗建立的單重PCR方法簡捷、特異煙在、敏感，适合基層獸醫實驗室對(duì)犬布作在魯氏菌病的檢測與鑒定。Development of a朋文 Single PCR Assay f他唱or Detection of Fo兒機ur Kinds of Brucella in Dog懂對sIn order to establ土理ish a single PCR assay appr票下opriate for detection of brucel裡現la in dogs，one pair of prim什科ers were designed through compar地少ison of genomic DNA sequences between r站子ough phenotype（B. canis）and smoo又長th phenotype（B. 紅北melitensis，B. abortus的了 and B. suis）strains，after opt筆作imization of reaction conditions，a PCR了都 assay was established for prelimin音房ary detection of th畫花e four kinds of brucella，then its sp空頻ecificity and sensitivity were evalu愛章ated，and 20 Brucella-seropositi腦區ve blood samples from dogs we一海re tested clinically. The resul個還ts showed that，for B. melitensis，B影林. abortus and B. suis，the fra玩亮gment with the length of 717 b子來p could be amplified理歌 by the established method，and for東國 B. canis，the fragment with 多時the length of 366 bp c店那ould be amplified；the lowest g鄉紅enomic DNA concentrations of B. canis空森，B. melitensis，B. s長多uis and B. abortus we為風re 5.07×10-2，6.20著作×10-2，7.80×10-3 and 5理票.50×10-2 ng/μL，res下老pectively；three positive samples fo街錢r B. canis were detec低看ted out in 20 blood samples，whi坐他ch was consistent with t市從he PCR results of usi業答ng GB/T 18646—2018 primers. As a co機街nclusion，the establ內場ished assay was appropriate for de場他tection of brucella in dogs in field ve家分terinary laboratories due to its 鐵為advantages of simplicity，specificity a煙靜nd sensitivity.全文下載鏈接：https://kns.cnk哥路i.net/kcms/detail/deta開河il.aspx?dbcode=CJFD&dbname=CJFDAUT木這O&filename=ZGDW202011016&v=B弟但h8a8ZT44%25mmd2BV4D1fobHDy%25mmd2BF74多答F2eWRbGd1dXZ%25mm朋地d2FFC%25mmd2FuzaTjuc0l4HBh%25mmd2B吧好ouBJZqFqPU
2020-11-06
全國(guó)非洲豬瘟防控專題巡從上回講座在哈爾濱啓動
關于增補筆試資格人員名單的通知
于康震在廣東江蘇督導檢查時(shí)強調：壓實屬地友讀責任 健全長(cháng)效機制 做好(hǎo)農村假冒僞劣食品整治和違法金讀屠宰行爲排查整治
中國(guó)新任OIE代表黃保續率團到為參加OIE第87屆大會(huì)
“科學(xué)使用獸用抗菌藥”百千萬數嗎接力公益再行動在泰州啓動
農業農村部辦公廳關于公布2019年全國(guó呢如)獸用抗菌藥使用減量化行動試點養殖場名單的通知
廣東省首例非洲豬瘟疫情的診斷與疫點消毒效果評價
韓長(cháng)賦會(huì)見出席二十國(guó)集團農業部長錯放(cháng)會(huì)議多國(guó)農業部長(cháng)
韓長(cháng)賦出席2019年二十國(guó)集團農業部長(ch好放áng)會(huì)議
關于2019年度公開(kāi)招聘面(事村miàn)試工作有關事(shì)項的通知
