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為準確檢測豬圓環(huán)病毒2型(PCV2)并分析其基因序列情況,在序列比對分析的基礎(chǔ)上,設(shè)計一對簡并引物和一條探針,經(jīng)體系優(yōu)化建立了PCV2的熒光PCR方法。該方法可檢出10拷貝的PCV2質(zhì)粒DNA,不與豬瘟病毒等多種病毒發(fā)生交叉反應(yīng)。對送檢的2批次45份病死豬臟器進行檢測,檢出25份陽性樣品,與套式PCR方法的復(fù)合率為95.6%;對3份強陽性樣品使用一對全基因組擴增引物,擴增了PCV2型全基因片段;經(jīng)克隆測序后分析,發(fā)現(xiàn)3個病毒基因分別屬于2種基因亞型(PCV2b和PCV2d)。本研究對于PCV2的準確檢測及其變異特征的了解具有參考意義。
Detection ofPorcine Circovirus Type 2 in Dead Pigs and Analysis on ItsComplete Genome Sequence
In order toaccurately detect porcine circovirus type 2(PCV2)and to analyze its genome sequence,a pair of degenerate primers and aprobe were designed based on sequence alignment analysis,after systematic optimization,a real-time PCR method was established,which could detect 10 copies ofplasmid DNA of PCV2,with no anycross reaction with classical swine fever virus and other viruses. Then twobatches of 45 organ samples collected from dead pigs were tested by establishedmethod,and 25 samples were detected to bepositive,the coincidence rate with nested-PCRwas 95.6%;for the three strong-positive samples,their complete gene fragments of PCV2were amplified by a pair of primers,then weresequenced and analyzed,and it wasfound that the genes of the three viruses were classified into two differentsubtypes(PCV2b and PCV2d). Therefore,it was of certain significance toconfirm relevant test results and recognize any variation of PCV2 by the abovemethod.
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國家獸藥產(chǎn)業(yè)技術(shù)創(chuàng)新聯(lián)盟 National veterinary drug industry technology innovation alliance |
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