RAPID DIAGNOSIS OF HERPES-SIMPLEX ENCEPHALITIS BY NESTED POLYMERASE CHAIN-REACTION ASSAY OF CEREBROSPINAL-FLUID
RAPID DIAGNOSIS OF HERPES-SIMPLEX ENCEPHALITIS BY NESTED POLYMERASE CHAIN-REACTION ASSAY OF CEREBROSPINAL-FLUID
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DOI：
10.1016/0140-6736(91)92155-u
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发表时间：
1991-01-26
期刊：
影响因子：
168.9
通讯作者：
FORSGREN, M
中科院分区：
文献类型：
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作者：
AURELIUS, E;JOHANSSON, B;FORSGREN, M
为了改善单纯疱疹脑炎的早期诊断，开发了一种采用两个“嵌套”引物对的聚合酶链反应 (PCR) 检测方法，用于扩增脑脊液 (CSF) 中的单纯疱疹病毒 DNA。 Southern印迹用于确认扩增的特异性。 该检测适用于来自 43 名连续单纯疱疹脑炎患者的 151 个脑脊液样本，通过在脑活检样本或尸检样本中发现单纯疱疹病毒/病毒抗原 (13) 和/或鞘内产生针对该病毒的 IgG 抗体 (40) 进行验证。 作为对照，对来自 60 名急性发热性局灶性脑病（最初怀疑是单纯疱疹性脑炎，但因鞘内抗体合成不存在而排除）患者的 87 份脑脊液样本进行了检测。 43名确诊单纯疱疹脑炎患者中，有42名通过PCR检测出单纯疱疹病毒DNA；在第一次采集的脑脊液样本中，除 1 例外，其余所有样本均呈阳性。 1 名 PCR 阴性患者在症状出现 20 小时后开始接受阿昔洛韦治疗。 所有对照受试者以及 270 名内部污染对照者均为 PCR 阴性。 在神经系统症状出现后 27 天抽取的样本中，PCR 结果仍然呈阳性。 该方法是一种快速、无创诊断单纯疱疹脑炎的手段；它具有高度的敏感性和特异性。
With the aim of improving early diagnosis of herpes simplex encephalitis a polymerase chain reaction (PCR) assay with two "nested" primer pairs was developed for the amplification of herpes simplex virus DNA in cerebrospinal fluid (CSF). Southern blotting was used to confirm the specificity of the amplification. The assay was applied to 151 CSF samples from 43 consecutive patients with herpes simplex encephalitis verified by the finding of herpes simplex virus/viral antigen in a brain biopsy sample or at necropsy (13) and/or intrathecal production of IgG antibody to the virus (40). As controls, 87 CSF samples from 60 patients with acute febrile focal encephalopathy (initially suspected to be herpes simplex encephalitis but excluded by the absence of intrathecal antibody synthesis) were tested. PCR detected herpes simplex virus DNA in 42 of the 43 patients with proven herpes simplex encephalitis; all but 1 were positive in the first CSF sample taken. The 1 PCR-negative patient had been treated with acyclovir from 20 h after the onset of symptoms. All the control subjects were PCR negative, as were 270 internal contamination controls. The PCR result remained positive in samples drawn up to 27 days after the onset of neurological symptoms. This method is a rapid and non-invasive means to diagnose herpes simplex encephalitis; it is highly sensitive and specific.