一株猫艾滋病病毒的巢式PCR检测及亚型鉴定

Nested PCR detection and subtype identification of one strain of Feline immunodeficiency virus

为了进一步了解猫免疫缺陷病毒(FIV)的区域流行情况,试验采用巢式PCR方法对15份流浪猫全血样品进行FIV gag基因扩增,检测出FIV阳性样品后再通过巢式PCR扩增得到FIV env基因V3~V5高变区序列,应用DNAMAN软件对阳性样品env基因的V3~V5高变区序列与NCBI中收录的参考毒株进行比对,使用MEGA 7.0软件构建进化树并进行遗传进化分析。结果表明:检测出1份FIV阳性样品,将FIV阳性株命名为XM07毒株,该毒株的env基因V3~V5高变区序列与NCBI中C78毒株的核苷酸序列相似性达到97.57%,与SH-2363毒株的核苷酸序列相似性达到96.83%;XM07毒株与SH-2363毒株在同一进化分支中,鉴定为A亚型。说明试验鉴定的FIV毒株未发生明显变异。

In order to further understand the regional prevalence of Feline immunodeficiency virus(FIV),in the test,nested PCR was used to amplify the FIV gag gene of 15 whole blood samples from stray cats;after detecting the FIV-positive samples,the sequences of the highly variable region of the FIV env V3-V5 region were then amplified by nested PCR.DNAMAN software was used to compare the V3-V5 highly variable region sequences of the env gene of the positive samples with the reference strains listed in NCBI,and MEGA 7.0 software was used to construct an evolutionary tree and conduct genetic evolution ary analysis.The results showed that one FIV-positive sample was detected,and the FIV-positive strain was named XM07 strain.The sequence of the highly variable region of env gene V3-V5 of this strain showed 97.57%nucleotide similarity with the C78 strain in NCBI and 96.83%nucleotide similarity with the SH-2363 strain;the XM07 strain was in the same evolutionary branch with the SH-2363 strain and was identified as subtype A.The results suggested that there was no obvious mutation of the FIV strain identified in the test.