阜阳市疑似恙虫病患者和恙螨及恙虫病鼠类中东方体分离及基因分型研究  

Orientia tsutsugamushi isolated from patients with suspected tsutsugamushi disease,chigger mites and mice with scrub typhus in Fuyang and genotypes

作　　者：于昌军 刘运喜[2] 李国兰[3] 张威[3] 卜戈[3] 孙良[3] 汪春新 YU Chang-jun , LIU Yun-xi, LI Guo-lan, ZHANG Wei, BU Ge, SUN Liang, WANG Chun-xin(Affiliated Fuyang Hospital of Anhui Medical University, Fuyang, Anhui 236000, Chin)

机构地区：[1]安徽医科大学附属阜阳医院检验科,安徽阜阳236000 [2]中国人民解放军总医院感染管理与疾病控制科,北京100039 [3]阜阳市疾病预防控制中心,安徽阜阳236030

出　　处：《中华医院感染学杂志》2018年第6期837-840,共4页Chinese Journal of Nosocomiology

基　　金：国家重点研发计划基金资助项目(2017YFC0806308); 总后应用基础研究重点资金资助项目(BWS13J027)

摘　　要：目的分析从患者全血、恙螨、野鼠肝脾等标本中分离的恙虫病东方体,为进一步确立阜阳市为新疫源地提供依据。方法采集2009年-2010年每年9月-11月辖区内疑似患者抗凝血、野鼠、恙螨标本,利用小白鼠传代分离法,分离恙虫病东方体,通过巢式PCR方法检测,对其测序结果进行基因分型。结果患者新鲜血液或抗凝血标本在传至3代~4代以后,小白鼠肝脾组织标本核酸检测结果部分为阳性(全血PCR阳性59组,经小白鼠分离后15株阳性;恙螨20组,其中3组PCR阳性,经小白鼠分离后1株阳性;野鼠256只,其中7只PCR阳性,经小白鼠分离35组,其中1株阳性),取扩增产物进行序列分析,证实为恙虫病东方体kawasaki基因型。结论首次从秋冬型恙虫病东方体流行季节患者、野外优势鼠种黑线姬鼠以及小盾纤恙螨体内分离的病原体为恙虫病东方体,进一步从病原学上证实阜阳市为新的疫源地。OBJECTIVE To identify the Orientia tsutsugamushi strains isolated from whole blood specimens of patients,chigger mites and liver and spleen specimens of field mice so as to provide evidence for further confirming that Fuyang is a new epidemic foci.METHODS From September to November in 2009-2010,the specimens were collected from anticoagulants of patients with suspected tsutsugamushi disease,wild mice and chigger mites,the O.tsutsugamushi strains were isolated by using mouse passaged separation method,and the genes were sequenced with the use of nested-PCR.RESULTS The nucleic acid test showed positive for part of the liver and spleen tissues of the mice as the fresh blood or anticoagulants specimens passed to third-fourth generation whole blood PCR positive group 59,15 strains were positive after the separation of the mice;the chigger mite 20 group,three of which were PCR positive,and 1 strain was positive after the separation of the mice;of the 256 wild mice,7 were PCR positive,the mice were separated to 35 groups,and 1 strain showed positive.The amplified products were sequenced,and the sequencing analysis showed that O.tsutsugamushi belonged to Kawasaki genotype.CONCLUSION The pathogen that is initially isolated from the autumn-winter tsutsugamushi epidemic seasonal patients,wild-type rodent Apodemus agrarius and small shield fiber mite body is the O.tsutsugamushi,and Fuyang is etiologically confirmed as the new epidemic foci.

关 键 词：恙虫病东方体 巢式聚合酶链反应 序列分析 疫源地 分离 

分 类 号：R376[医药卫生—病原生物学]