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目的了解云南省德钦县犬感染嗜吞噬细胞无形体的状况。方法对2013-2014年期间在德钦县采集的315份犬血标本,应用巢式聚合酶链反应进行嗜吞噬细胞无形体16S rRNA基因扩增和测序分析,并将所测序列与Gen Bank中注册的基因序列进行同源性和进化树分析。结果 315份犬血样本中共有18份扩增到目的片段,总阳性率为5.71%(18/315),其中中华田园犬的阳性率为7.14%(12/168),藏犬的阳性率为5.13%(4/78),哈巴犬的阳性率为2.99%(2/67),狼犬中未检出HGA。所检出的嗜吞噬细胞无形体16S rRNA基因与Gen Bank中收录的部分嗜吞噬细胞无形体16S rRNA基因序列的同源性高达99%~100%。进化树分析结果显示,当地无形体分为2类流行株,将其命名为Yunnan HGA1和Yunnan HGA2,分别占检出菌株的16.67%和83.33%。Yunnan HGA1与在乌拉圭、西伯利亚和远东地区发现的全沟硬蜱中检测到的无形体片段(HM366583)同源性为100%,Yunnan HGA2与加拿大发现的肩突硬蜱中检测到的无形体片段(HG916766)同源性为99%。结论云南省德钦地区犬存在嗜吞噬细胞无形体的感染,进一步开展相应媒介、宿主及人群感染调查十分必要。
Objective To understand the status of phagocytosis of phagocytes in dogs in Deqin County of Yunnan Province. Methods 315 canine blood samples collected from Deqin County during 2013-2014 were amplified and sequenced by Nested Polymerase Chain Reaction (PCR) for 16S rRNA gene of phagocyte phage, and the sequences were compared with Gen Bank Registered gene sequence homology and evolutionary tree analysis. Results A total of 18 out of 315 samples of canine blood samples were amplified. The total positive rate was 5.71% (18/315). The positive rate of Chinese pastoral dogs was 7.14% (12/168). The positive rate of Tibetan dogs was 5.13% (4/78), Pug’s positive rate was 2.99% (2/67), HGA was not detected in dogs. The homology between the 16S rRNA gene of phagocytic phagocytosis and the 16S rRNA gene sequence of some phagocytes in Gen Bank was as high as 99% ~ 100%. The results of phylogenetic tree analysis showed that the two species were classified into two types of endemic: Yunnan HGA1 and Yunnan HGA2, accounting for 16.67% and 83.33% of the isolates respectively. Yunnan HGA1 has 100% identity to the invisible fragment (HM366583) detected in Ivus tubers found in Uruguay, Siberia and the Far East. Yunnan HGA2 has been identified as an invisible fragment detected in Ixodes scapularis in Canada (HG916766) with 99% homology. Conclusion Infection of phagocytic phagocytes in dogs in Deqin area, Yunnan Province, may be necessary to further investigate the infection of host, host and host.