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一项关于聚合酶链反应检测提交至临床病毒学实验室的脑脊液中单纯疱疹病毒的前瞻性研究。

A prospective study of the polymerase chain reaction for detection of herpes simplex virus in cerebrospinal fluid submitted to the clinical virology laboratory.

作者信息

Aslanzadeh J, Osmon D R, Wilhelm M P, Espy M J, Smith T F

机构信息

Department of Laboratory Medicine, Mayo Clinic, Rochester, MN 55905.

出版信息

Mol Cell Probes. 1992 Oct;6(5):367-73. doi: 10.1016/0890-8508(92)90029-w.

Abstract

Polymerase chain reaction (PCR) was prospectively performed with cerebrospinal fluid (CSF) from 51 patients whose CSF was available for analysis and was submitted for viral culture and/or herpes simplex virus (HSV) serology and 20 patients whose CSF was submitted exclusively to the Clinical Biochemistry Laboratory. Primers were used that flanked a 92 bp segment of the HSV DNA polymerase gene (35 cycles). Amplified products were electrophoresed on agarose gel, blotted onto nylon membrane, and probed with a 32P-labelled sequence internal to the primers. For nested PCR, 1 microliter of PCR product was amplified for an additional 35 cycles before electrophoresis and Southern blot analysis. Review of the clinical records revealed that 15 patients had central nervous system (CNS) infections. Specific HSV DNA sequences were detected in CSF specimens of three of the individuals [PCR(2), nested PCR(1)]. Two of these patients had disseminated HSV infection including encephalitis and one patient had aseptic meningitis. The diagnoses of the 12 patients with CNS infection who did not have HSV DNA detected in CSF included encephalitis [varicella-zoster virus (1), cytomegalovirus (1), Mycoplasma pneumoniae (1)], meningitis [Neisseria meningitidis (1), Coccidioides immitis (1), Enterovirus (1), aseptic meningitis (1)], varicella-zoster radiculitis (2), human immunodeficiency virus dementia (2), and transverse myelitis due to Epstein-Barr virus (1). Importantly, HSV DNA was also not detected in the CSF of the 36 patients who did not have CNS infection and 20 samples submitted exclusively to the Clinical Biochemistry Laboratory. Our findings demonstrate the utility of PCR as a rapid, non-invasive method for the routine laboratory diagnosis of CNS infection due to HSV.

摘要

对51例脑脊液可用于分析并已送检进行病毒培养和/或单纯疱疹病毒(HSV)血清学检测的患者以及20例仅将脑脊液送检至临床生物化学实验室的患者的脑脊液前瞻性地进行了聚合酶链反应(PCR)检测。所用引物位于HSV DNA聚合酶基因的一段92 bp片段两侧(35个循环)。扩增产物在琼脂糖凝胶上进行电泳,转移至尼龙膜上,并用引物内部的32P标记序列进行杂交。对于巢式PCR,在电泳和Southern印迹分析前,取1微升PCR产物再扩增35个循环。查阅临床记录发现,15例患者患有中枢神经系统(CNS)感染。在其中3例个体的脑脊液标本中检测到了特异性HSV DNA序列[PCR检测到2例,巢式PCR检测到1例]。其中2例患者患有播散性HSV感染,包括脑炎,1例患者患有无菌性脑膜炎。在脑脊液中未检测到HSV DNA的12例CNS感染患者的诊断包括脑炎[水痘-带状疱疹病毒(1例)、巨细胞病毒(1例)、肺炎支原体(1例)]、脑膜炎[脑膜炎奈瑟菌(1例)、粗球孢子菌(1例)、肠道病毒(1例)、无菌性脑膜炎(1例)]、水痘-带状疱疹神经根炎(2例)、人类免疫缺陷病毒痴呆(2例)以及由EB病毒引起的横贯性脊髓炎(1例)。重要的是,在36例无CNS感染的患者的脑脊液以及仅送检至临床生物化学实验室的20份样本中也未检测到HSV DNA。我们的研究结果证明了PCR作为一种快速、非侵入性方法用于HSV所致CNS感染的常规实验室诊断的实用性。

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