安徽地区家畜嗜吞噬细胞无形体16S rRNA基因检测和序列分析

Genetic identification and sequence analysis of the 16S rRNA Genes of Anaplasma phagocytophilum in Anhui Province,China

目的了解安徽农村地区散养家畜自然感染嗜吞噬细胞无形体状况和基因特征。方法应用巢式PCR对怀远、明光和广德县采集的148份家畜全血样本进行嗜吞噬无形体16S rRNA基因检测,并对扩增产物测序,应用生物软件MEGA4.0进行序列分析。结果 148份家畜全血样本中检测出山羊、牛和犬嗜吞噬细胞无形体16S rRNA基因阳性率分别22.1%、25.0%和40.0%。26份PCR阳性扩增产物测序成功,序列分析结果表明：检出序列可归为4群,I群优势株占分析序列的66.7%,与北京、浙江和湖北等地检测序列同属一个分支。目的基因测定序列与参考序列同源性达97.0~99.0%。结论安徽调查地区农村家畜动物存在嗜吞噬细胞无形体感染,山羊是该地区嗜吞噬细胞无形体的重要宿主动物之一。

Objective To understand the status and gene characteristics of Anaplasma phagocytophilum infection among domestic animals in the rural area of Anhui province,China.Methods Nest-PCR was applied in the study to detect 148 blood samples of domestic animals,including 138 goats,12 dogs and 5 ox,from Guangde,Mingguang and Huaiyuan county in Anhui province.The positive PCR products were sequenced.Clustal X and Mega4.0 bio-software were used respectively to do nucleotide alignment and phylogenetic tree based on 16S rRNA genes.Results The positive rate of Anaplasma phagocytophilum were 22.1%,25.0% and 40.0% respectively among goats,dogs and ox.26 PCR products were sequenced successively.Sequence analysis results showed that the detected sequences could be classified into 4 groups,and group I strains accounted for 66.7%.The nucleotide sequences homologies of the partial 16SrRNA were 97.0%~99.0%between the objective and reference sequences.Conclusion The prevalence rate of Anaplasma phagocytophilum were identified in the rural areas of Anhui province,and goats were important hosts of Anaplasma phagocytophilum.