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用于诊断淋巴细胞性脉络丛脑膜炎病毒感染的逆转录-聚合酶链反应检测方法的开发及其在一项前瞻性监测研究中的应用。

Development of a reverse transcription-polymerase chain reaction assay for diagnosis of lymphocytic choriomeningitis virus infection and its use in a prospective surveillance study.

作者信息

Park J Y, Peters C J, Rollin P E, Ksiazek T G, Gray B, Waites K B, Stephensen C B

机构信息

Department of International Health, School of Public Health, University of Alabama at Birmingham 35294, USA.

出版信息

J Med Virol. 1997 Feb;51(2):107-14.

PMID:9021540
Abstract

Lymphocytic choriomeningitis virus (LCMV), which is one of several arenaviruses that are pathogenic for humans, causes encephalitis and meningitis in man. In this study, single-stage and nested reverse transcription-polymerase chain reaction (RT-PCR) assays were developed that targeted the GPC and N genes of LCMV. Both assays detected < 1 TCID50 unit of LCMV. These assays were used to measure the incidence of LCMV infection by testing cerebrospinal fluid (CSF) samples with > or = 10 leukocytes/microl collected over 1 year from patients undergoing lumbar puncture for diagnostic reasons at two Birmingham hospitals. Samples were tested for the presence of LCMV RNA by using the RT- PCR assay and for LCMV-specific IgM antibody by using an ELISA assay. None of the specimens collected from 813 patients was positive by either assay. Although no cases of acute infection were detected, 4% (11/272) of serum collected from a subset of patients was positive for LCMV-specific IgG. A significantly greater rate of seropositivity was found among subjects over 60 years of age (9.4%; P < 0.025) than was found in younger subjects (2.4% at 30-59 years of age; 0% at < 30 years of age). These data suggest that serious central nervous system disease due to LCMV infection is not common in this population. The high rate of seropositivity in those over 60 years of age suggest that infection was once more common.

摘要

淋巴细胞性脉络丛脑膜炎病毒(LCMV)是几种可致人类致病的沙粒病毒之一,可引起人类脑炎和脑膜炎。在本研究中,开发了针对LCMV的糖蛋白(GPC)基因和核蛋白(N)基因的单阶段和巢式逆转录聚合酶链反应(RT-PCR)检测方法。两种检测方法均能检测到<1个半数组织培养感染剂量(TCID50)单位的LCMV。通过对来自伯明翰两家医院因诊断原因接受腰椎穿刺的患者在1年多时间里收集的每微升白细胞数≥10个的脑脊液(CSF)样本进行检测,使用这些检测方法来测定LCMV感染的发生率。通过RT-PCR检测方法检测样本中LCMV RNA的存在情况,并通过酶联免疫吸附测定(ELISA)检测方法检测LCMV特异性IgM抗体。从813例患者收集的标本中,两种检测方法均未检测到阳性结果。虽然未检测到急性感染病例,但从一部分患者中收集的血清有4%(11/272)LCMV特异性IgG呈阳性。60岁以上的受试者中血清阳性率(9.4%;P<0.025)显著高于年轻受试者(30至59岁为2.4%;<30岁为0%)。这些数据表明,在该人群中,由LCMV感染引起的严重中枢神经系统疾病并不常见。60岁以上人群中血清阳性率较高,表明既往感染更为常见。

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