【摘要】：目的分析3株經(jīng)野狼-家犬-牦牛傳播的狂犬病病毒N基因部分核酸序列,以確定其傳染源和傳染途徑。方法分離發(fā)病動(dòng)物的RNA進(jìn)行DFA檢測(cè)和RT-PCR檢測(cè),陽(yáng)性者進(jìn)行N基因部分編碼區(qū)核苷酸測(cè)序檢測(cè),采用ATCG ver4軟件進(jìn)行序列拼接,用DNAMAN將所測(cè)序列N基因進(jìn)行核苷酸序列同源性比對(duì),用ClustalX軟件Bootstrap N-J Tree法構(gòu)建系統(tǒng)發(fā)生樹(shù)。結(jié)果發(fā)病宿主動(dòng)物腦組織標(biāo)本和脊髓經(jīng)DFA檢測(cè)到內(nèi)基氏小體和病毒顆粒,RT-PCR擴(kuò)增狂犬病毒核酸陽(yáng)性,擴(kuò)增產(chǎn)物大小為256bp,與目標(biāo)片段大小一致。RT-PCR擴(kuò)增產(chǎn)物經(jīng)純化后測(cè)序,N基因核苷酸序列3毒株間的同源性為99.28%~99.73%,推導(dǎo)的氨基酸序列同源性為99.46%~100.00%,牦�？袢《竞塑账嵝蛄信c咬傷犬同源性99.28%,在自斃狼和病犬同源性為100%,牦�？袢《竞塑账嵝蛄信c本室2012年測(cè)序的瑪多犬N基因核苷酸同源性99.7%,與2012年內(nèi)蒙古野生貍neimeng102B(EU652445.1)的同源性為95.28%,與2008年USA994-dog(EF611868.1)的同源性為95.94%,與2009BeijingHu1(EU700031.1)的同源性為85.9%。3株狂犬病毒株間N基因變異較小,穩(wěn)定性較高,病毒核苷酸序列均為基因China IV,具有一定的地域性,進(jìn)化分析顯示均屬于與野生動(dòng)物最為相關(guān)的北極群。結(jié)論狂犬病可經(jīng)野狼-家犬-家養(yǎng)牦牛傳播,本起疫情中野狼為第一傳染源。
[Abstract]:Objective to analyze the partial nucleic acid sequence of rabies virus N gene transmitted by wolf, domestic dog and yak, and to determine the source of infection and route of transmission. Methods the RNA of the diseased animals were detected by DFA and RT-PCR, the nucleotides in the partial coding region of N gene were sequenced, and the sequences were spliced by ATCG ver4 software. The nucleotide sequence homology of the N gene was compared by DNAMAN, and the phylogenetic tree was constructed by ClustalX software Bootstrap N 鈮，

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