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目的了解云南地区鼠类和人自然感染恙虫病东方体的情况。方法采用鼠夹和鼠笼捕鼠,从鼠类脾脏及人血中提取DNA,应用巢式聚合酶链反应扩增恙虫病东方体(Ot)sta56基因,并进行基因序列测定分析。结果捕获小型哺乳动物7属11种485只,收集到不明原因发热病例血液样本23份。从动物脾脏样本中检出Ot sta56基因片段2份(HHSP65和HHSP112),阳性率0.41%;序列分析显示两者同源性100%,而与中国台湾TT0711a株的同源性最高为89.6%,与日本Kawasaki株的同源性为79.7%;进化树分析显示HHS65与TT0711a株不在同一分支上。从不明原因发热患者血液样本中检出Ot sta56基因片段1份(DLRX9),阳性率4.35%;序列分析显示该序列与泰国PG9和Li3b株及中国浙江Jin/2012株的同源性最高,均为98.4%,与Karp株的同源性为81.9%,与HHSP65株的同源性64.8%;进化树显示DLRX9与泰国PG9和Li3b、中国浙江Jin/2012株在同一分支上。结论病原学初步证实引发云南地区恙虫病流行的Ot不仅有Karp基因型Ot,还有新型Ot,该新型Ot可能为日本Kawasaki类似株。
Objective To understand the natural infection of rodents and oriental rodents in rodents and humans in Yunnan Province. Methods DNA was extracted from mouse spleen and human blood using mouse and mouse cage traps, and sta56 gene was amplified by nested polymerase chain reaction and sequenced. Results A total of 485 small mammals (7 genera and 11 species) were captured, and 23 blood samples were collected from patients with unexplained fever. Two copies of Ot sta56 gene fragment (HHSP65 and HHSP112) were detected in animal spleen samples, with a positive rate of 0.41%. Sequence analysis showed 100% homology between the two genes and the highest homology with TT0711a strain in Taiwan of China was 89.6% The homology with Kawasaki strain in Japan was 79.7%. Phylogenetic tree analysis showed that HHS65 and TT0711a strains were not on the same branch. One (DLRX9) gene fragment of Ot sta56 gene was detected in the blood samples of patients with unexplained fever and the positive rate was 4.35%. Sequence analysis showed that the sequence shared the highest homology with PG9 and Li3b strains in Thailand and Jin / 2012 strain in Zhejiang Province, both of which Was 98.4%, which shared 81.9% identity with Karp strain and 64.8% with HHSP65 strain. The phylogenetic tree showed that DLRX9 was on the same branch with PG9 and Li3b in Thailand and Jin / 2012 strain in Zhejiang Province, China. Conclusions The etiology initially confirmed that Ot, which induces the tsutsugamushi epidemic in Yunnan, not only has the Karp genotype Ot but also the new Ot, which may be a similar strain of Kawasaki in Japan.