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北京林区莱姆病基因型自然疫源地及其病原体的聚合酶链反应探索性研究

[Exploratory study on natural focus and its causative agent of genotype of Lyme disease by polymerase chain reaction in the forest areas of Beijing].

作者信息

Li Jianmin, Cao Wuchun, Zhang Xitan, Wu Xiaoming, Zhang Panhe, Zhao Qiumin, Yang Hong, Dong Zhenying, Cai Shengli

机构信息

Institute of Microbiology and Epidemiology of Academy of Military Medical Science, Beijing 100071, China.

出版信息

Zhonghua Liu Xing Bing Xue Za Zhi. 2002 Jun;23(3):209-12.

Abstract

OBJECTIVE

To learn the existence of natural focus of Lyme disease and its distribution.

METHODS

A semi-nested polymerase chain reaction (PCR) method was developed for detection and genotyping of Borrelia burgdorferi on basis of outer surface protein A (OspA) gene. Ticks and mice collected from 6 forest areas in Beijing were detected with above methods. The positive PCR products were cloned and sequenced. The sequences were compared with published sequences for homology. IFA as used to detect IgG antibody on Borrelia burgdorferi. Lyme disease spirochete were isolated from H. longicornis were also attempted.

RESULTS

B. Burgdorferi sensu lato were detected from 939 ticks and 250 mice specimens collected from above 6 study sites using primer pairs OA(1)/OA(4) and SL/OA(4). Only the specimens collected from Dongling mountain showed positive amplification. One in three adult Ixodes persulcatus with one of 57 nymph Ixodes persulcatus showed positive while 9 of 119 (7.56%) mice specimens showed positive, of which 8 were B. grinii and one B. afzelii. In this study, we attempted to isolate B. burgdorferi sensu lato strains from 160 H. longicornis ticks (20/group) but failed. Serological survey showed a 9.1% (5/55) infection rate with B. burgdorferi sensu lato in the mice of Dongling mountain forest areas.

CONCLUSIONS

The natural focus of Lyme disease including B. garinii and B. afzelii might have existed in Dongling mountain of Mentougou district, Beijing. Ixodes persulcatu and mice may serve as vectors and reservoirs, respectively.

摘要

目的

了解莱姆病自然疫源地的存在情况及其分布。

方法

基于外表面蛋白A(OspA)基因,建立一种半巢式聚合酶链反应(PCR)方法用于检测伯氏疏螺旋体并进行基因分型。采用上述方法对从北京6个林区采集的蜱和小鼠进行检测。将PCR阳性产物进行克隆和测序,并与已发表序列比较同源性。采用间接免疫荧光法(IFA)检测伯氏疏螺旋体IgG抗体。还尝试从长角血蜱中分离莱姆病螺旋体。

结果

使用引物对OA(1)/OA(4)和SL/OA(4),从上述6个研究地点采集的939只蜱和250只小鼠标本中检测到伯氏疏螺旋体狭义种。仅从东灵山采集的标本显示阳性扩增。3只成年全沟硬蜱中的1只和57只若虫全沟硬蜱中的1只呈阳性,而119只小鼠标本中有9只(7.56%)呈阳性,其中8只为格林尼亚伯氏疏螺旋体,1只为阿氏疏螺旋体。在本研究中,尝试从160只长角血蜱(每组20只)中分离伯氏疏螺旋体狭义种菌株,但未成功。血清学调查显示,东灵山森林地区小鼠中伯氏疏螺旋体狭义种的感染率为9.1%(5/55)。

结论

北京门头沟区东灵山可能存在包括伽氏伯氏疏螺旋体和阿氏疏螺旋体在内的莱姆病自然疫源地。全沟硬蜱和小鼠可能分别作为传播媒介和宿主。

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