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目的从分子水平探讨山东地区不同季节优势恙螨自然感染恙虫病东方体(Ot)情况及其作为恙虫病传播媒介的可能性。方法根据Ot-Sta56 kDa外膜蛋白基因部分序列设计Ot种和型特异性引物,对标本提取的DNA先采用种特异性引物扩增,然后再采用型特异性引物扩增分型,并对部分标本进行序列测定。结果从不同季节优势恙螨幼虫共得到11株Ot分离株和16份恙螨匀浆,从这27份标本提取的DNA经首轮PCR扩增后,18份标本有预期的目的带。对首轮PCR产物采用nested PCR扩增分型,结果17份为Kawasaki型,1份(LHGM2株)为Karp型。序列同源性分析结果证实了nested PCR分型结果:XDM2株首轮PCR产物碱基序列与。Kawasaki型相应DNA片段的碱基序列同源性最高,为97．00％,在系统发育树上与Kawasaki株位于同一分支,应属于Kawasaki型; LHGM2株首轮PCR产物碱基序列与Karp型相应DNA片段的碱基序列同源性最高,为96．45％,在系统发育树上与Karp株位于同一分支,应属于Karp型。结论山东地区不同季节优势恙螨中存在Ot自然感染,Ot主要基因型为Kawasaki型。
Objective To explore the natural status of Otomycosis tsutsugamushi (Ot) and its possibility as vectors of tsutsugamushi disease in different seasons in Shandong province. Methods According to Ot-Sta56 kDa Outer Membrane Protein gene partial sequence design Ot species and type specific primers, DNA samples extracted using species-specific primer amplification, and then use type-specific primers to amplify the type, and part of the Specimens were sequenced. Results 11 Ot isolates and 16 chigger mites homozygotes were obtained from chigger mite larvae in different seasons. The DNA extracted from these 27 samples was amplified by the first round of PCR, and 18 samples had the expected target band. The first round of PCR products using nested PCR amplification typing, the results of 17 Kawasaki type, 1 (LHGM2 strain) Karp type. Sequence homology analysis confirmed the nested PCR typing results: XDM2 first-pass PCR product base sequence and. The Kawasaki-type DNA fragment had the highest homology of 97.00%, which belonged to Kawasaki type in the phylogenetic tree and Kawasaki strain. The first PCR product of LHGM2 had the same base sequence as Karp-type The nucleotide sequence of the DNA fragment has the highest homology of 96.45%. It belongs to the Karp type in the same branch of Karp strain in phylogenetic tree. Conclusion There is natural infection of Ot in chigger mites in different seasons in Shandong Province. The main genotype of Ot is Kawasaki type.