山东地区不同季节优势恙螨自然感染恙虫病东方体的分子流行病学调查

张倩; 刘运喜; 高媛; 赵仲堂; 张景兰; 杨占清; 薛健; 曹务春

Abstract

张倩,刘运喜,高媛,赵仲堂,张景兰,杨占清,薛健,曹务春.山东地区不同季节优势恙螨自然感染恙虫病东方体的分子流行病学调查[J].Chinese journal of Epidemiology,2006,27(7):600-603

山东地区不同季节优势恙螨自然感染恙虫病东方体的分子流行病学调查

Study on the molecular epidemiology regarding the natural infection of Orientia tsutsugamushi in 4 species of dominant chiggers collected in various seasons from the foci of Shandong province

Received:October 21, 2005

DOI：

KeyWord: 恙虫病恙螨恙虫病东方体 Sta56基因

English Key Word: Scrub typhus Chigger mite Orientza tsutsu}amushi Sta56 gene

FundProject:国家自然科学基金资助项目(30371237);解放军总后勤部卫生部全军医药卫生科研基金资助项目(01Q016)

Author Name	Affiliation	E-mail
ZHANG Qian	State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing 100071, China
LIU Yun-xi	State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing 100071, China
GAO Yuan	济南军区联勤部疾病预防控制中心
ZHAO Zhong-tang	山东大学公共卫生学院
ZHANG jing-lan	济南军区联勤部疾病预防控制中心
YANG Zhan-qing	济南军区联勤部疾病预防控制中心
XUE Jian	济南军区联勤部疾病预防控制中心
CAO Wu-chun	State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing 100071, China	aowc@nic.bmi.ac.cn

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Abstract:

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。

English Abstract:

Objective In order to investigate the natural infection of Orientia tsutsugamushi (Ot) in 4 species of dominant chiggers collected in various seasons from the foci of Shandong province. Methods Species-specific and type-specific primers were designed according to the published sequence of Sta56 gene. The DNAs extracted from the samples were amplified by initial PCR using species-specific primers before the genotypes of initial PCR amplicons were identified by nested PCR using type-specif ic primers, and using the nucleotide sequence analysis of 2 representative samples to confirm the PCR Results. Results The expected specific fragments from 18 of the 27 Ot-Sta56 genes (11 isolated strains and 16 homogenates of various mite larvae) were initially amplified by PCR. Out of the 18 initial PCR positive samples, 17 were identified as Kawasaki types by nested PCR, while one LHGM2 strain was identified as Karp type. DNA sequence analysis confirmed the nested PCR Results. The Sta56 gene nucleotide sequence homology was 97.00% to Japan Kawasaki strain of XDM2 from the above 17 samples which were identified as Japan Kawasaki strain by nested PCR. The sequence homology to Karp strain of LHGM2 was 96.45 %. Conclusion These data indicated that Ot was widely distributed in 4 species of dominant chigger mites collected from different seasons in Shandong province. The epidemic genotypes of Ot belonged to Kawasaki strain.

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