新疆革蜱源性牛无浆体的PCR检测

PCR Detection of Dermacentor-borne Anaplasma bovis in Xinjiang

为了解新疆部分地区优势媒介革蜱类传播牛无浆体的状况,随机采集了吐鲁番、巴州等牛场和放牧牛身上寄生的革蜱(N=140只),以16S rRNA基因为靶标分别设计了无浆体属通用引物(EE1/EE2)和牛无浆体特异性引物(AB1f/AB1r),运用嵌套式PCR方法扩增其全基因组DNA。试验结果显示,在140只革蜱全基因组DNA中,扩增出阳性条带的13只,其牛无浆体的阳性率为9.3%(13/140),这结果被阳性革蜱同一个群的革蜱动物传播试验所验证。经阳性病原DNA的克隆测序、比对后,发现该牛无浆体病原DNA序列与数据库中的西藏株(AF414399)和福建株(DQ324547)相似,其同源性均为99%。试验表明,新疆牛无浆体可能被当地的优势媒介革蜱所携带和传播,该参数可为地方蜱传无浆体病的综合防治提供科学依据。

In order to investigate whether Anaplasma bovis was spread mainly by Dermacentor in parts of Xinjiang,one hundred and forty ticks from catties were collected in Turpan,Bazhou,and other epidemic areas.The universal primers（EE1/EE2） and gene specific primers（AB1f /AB1r） were designed to target the 16 S rRNA of Anaplasma bovis and nested PCR was applied to amplify its full genomic DNA.The results showed that 13 positive bands were observed in 140 genome DNA samples from Dermacentor,and the positive rate of Anaplasma bovis was 9.3%（13/140）,which were verified by Dermacentor spread pathogenic on animals of experiments from the same group of positive Dermacentor.DNA-positive pathogens were cloned and sequenced.The results showed that the DNA sequence of Anaplasma bovis was similar with Tibet strain（AF414399） and Fujian strain（DQ324547）in the database,and their homology were 99%.The study suggestes that Anaplasma bovis is in mainly in Dermacentor and transmissed by Dermacentor in Xinjiang,our data provides the basis for the prevention of tick-borne Anaplasma in Xinjiang.