无菌性脑膜炎患者肠道病毒感染鉴定方法

崔连东; 李凡; 李哲; 易世红; 杨彤; 尹正

doi:10.11847/zgggws2005-21-12-40

无菌性脑膜炎患者肠道病毒感染鉴定方法

Enteroviral RT-PCR assay on diagnosis of enterovirus in aseptic meningitis patients

摘要

摘要:
目的建立一种快速、准确的鉴定无菌性脑膜炎患者肠道病毒感染的方法。并与传统的细胞培养及血清分型相比较。
方法对78例无菌性脑膜炎患儿脑脊液中肠道病毒, 用人宫颈癌细胞(HeLa)、横纹肌细胞(RD)和人喉癌细胞(Hep-2)细胞进行分离培养血清分型, 同时用根据肠道病毒5'UTR区基因序列设计引物, 用PCRT-R法扩增病毒的基因。
结果该方法可检测的灵敏度的组织培养感染量(TCID₅₀)为0.1;78例脑脊液(CSF)中46例肠道病毒培养为阳性, 而用RT-PCR法检测52例阳性。
结论该方法与细胞培养及肠道病毒血清分型法相比较具有较高的灵敏度和特异性, 可快速、灵敏地检测脑脊液中的肠道病毒。

Abstract:
Objective To compare the results of virus isolation with those of the method of RT-PCR in identical specimens from cases of suspected aseptic meningitis.Cerebrospinal fluid samples were collected for viral examination from 78 pediatric patients with suspected aseptic meningitis.
Methods These samples were inoculated in the cell lines of HeLa, RD and Hep-2 cell.The viral RNA was detected by RT-PCRmethod according to the region of 5 UTR.The isolated viruses were typed by neutralization test.
Results 46 of the 78 specimens were detected to be enter ovirus positive by culture method, while 52 of them yielded positive results when RT-PCRmethod was used.
Conclusion Because of higher sensitivity and rapidity of the method of RT-PCR, it is superior to isolate the virus from CSFfor the diagnosis of enterovirus meningitis.

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