用通用引物建立的RT-PCR诊断肠道病毒性脑炎及脑膜炎

Application of reverse transcription polymerase chain reaction based on picornaviridae-specific primer to diagnosis of enteroviral encephalitis and meningitis

目的 用通用引物建立逆转录聚合酶链反应（ＲＴ－ＰＣＲ），达到广谱、特异及快速诊断肠道病毒（ＥＶ）性脑炎及脑膜炎。方法 用针对ＥＶ基因组５’端非编码区保守区的１对通用引物建立的ＲＴ－ＰＣＲ检测多种ＥＶ标准血清型，同时检测病毒性脑膜炎和脑炎患儿的脑脊液。结果 此种ＲＴ－ＰＣＲ可检测４２种ＥＶ的标准血清型，所用毒株在１０－１ＴＣＩＤ５０时仍可见到１９４ｂｐ的阳性条带。检测了３２例病毒性脑膜炎和脑炎患儿的脑脊液中ＥＶ－ＲＮＡ，１８例为阳性（５６．３％）。ＥＶ－ＲＮＡ阳性患儿的昏迷、瘫痪、抽搐、ＣＳＦ－蛋白质及低密度灶数（ＣＴ）分别为４（２２．２％）、８（４４．４％）、１２（６６．７％）、４６２ｍｇ／Ｌ（ｘ）及４（２２．２％）；ＥＶ－ＲＮＡ阴性患儿的昏迷、瘫痪、抽搐、ＣＳＦ－蛋白质、及低密度灶数（ＣＴ）分别为１０（７１．４％）、６（４２．９％）、１２（８５．７％）、３８６ｍｇ／Ｌ（ｘ）及４（２８．８％），仅昏迷发生率有极显著差异（Ｐ＜０．０１）。结论 用通用引物建立的ＲＴ－ＰＣＲ诊断ＥＶ性脑膜炎和脑炎，具有广谱性、特异性、敏感性及简洁、快速的特点，诊断ＥＶ性脑膜炎和脑炎主要依靠病原学检查。

Objective To determine the feasibility of reverse transcription polymerase chain reaction( RT - PCR)based on picornaviridae- specific primer( PSP) in diagnosis of enteroviral encephaitis and meningitis. Methods RT - PCR with one pair of PSP against the conserved 5' noncoding region of enterovirus( LV) was employed to investigate enterovirus RNA from standard strains( SS) and patients' cerebral spinal fluid ( CSF) respectively. Results EV - RNA of 42 sorts of standard serotypes could be detected, positive band 194bp could still be observed at the leval of 10- 1TCID50 of SS. Positive findings were also shown in the CSF form 18/32(56.3%)cases with viral encephalitis and miningitis. The incidence of coma, paralysis, and seizure, level of protein in CSF and the number of low density in CT scanning in EV - RNA positive group were 4 (22.2%),8(44.4%),l2(66.7%),462mg/L(x)and4(22.2%),which in EV-RNA negative group were 10(71.4%),6 (42 9%), 12(85.7%), 386mg/L( x )and 4(28.8% )respectively. Only significant was the diffeference in incidence of coma between both of groups(P <0.01 .Concluslon RT - PCR with one pair of PSP against the conserved 5' noncoding region of LV is characterized by broad spectrum, sharp specificity and high sensitivity during study of EV - induced encephalitis and meningitis and the procedure saves trouble. The diagnosis of EV encephalitis and meningitis mainly depends on etiologies1 investigation.