摘要
目的 提高无菌性脑膜炎和脑炎病原早期诊断的敏感性和特异性。方法 分别设计针对肠道病毒(EV) 5’UTR、Ⅰ型和Ⅱ型单纯疱疹病毒 (HSVⅠ ,Ⅱ )糖蛋白D基因、人巨细胞病毒 (CMV)立即早期蛋白基因、EBV核抗原 1基因和肺炎支原体 (MP)ATPase操纵子基因的巢式PCR引物 ,应用巢式 聚合酶链反应 (nested PCR)技术检测无菌性脑膜炎和脑炎患儿的脑脊液标本中EV、Ⅰ型和Ⅱ型HSV、CMV、EBV及MP。结果 所用不同病原体的引物之间无交叉反应 ,特异性高。收集到的 15 8例脑脊液标本中有 10 2例 (6 4 6 % )检测到病原 ,包括 4 2例EV (42 / 15 8,2 6 6 % ) ,2 7例HSVⅡ (2 7/ 15 8,17 1% ) ,16例EBV (16 / 15 8,10 1% ) ,8例MP(8/ 15 8,5 1% ) ,6例CMV(6 / 15 8,3 8% ) ,3例HSVⅠ (3/ 15 8,1 9% )。这些阳性标本中有 10例检测到 2种或 3种病原。结论 PCR技术在无菌性脑膜炎和脑炎病原的早期诊断中有潜在的应用价值 ,可在中枢神经系统感染的病原鉴别中作为一种辅助性实验室诊断为临床诊治提供参考依据。
Objective To improve the sensitivity and specificity of the early pathogenic diagnosis of aseptic meningitis and encephalitis.Methods Nested-PCR primers specific to the 5'UTR of enterovirus (EV),glycoprotein D-gene of herpes simplex virus (HSV) Ⅰ and Ⅱ,immediate early antigen (IEA1)gene of human cytomegalovirus (CMV),gene of Epstein-Barr virus (EBV)nuclear antigen 1 (EBNA1) and ATPase operon gene of mycoplasma pneumonia (MP) were respectively designed.Cerebrospinal fluid samples were examined to detect EV,HSV Ⅰ and Ⅱ,CMV,EBV and MP in children with aseptic meningitis or encephalitis by using nested-PCR technique.Results This detecting method was specific,in that no cross reactions occurred between the primers of different antigens.Of the 158 cerebrospinal fluid samples,pathogens were detected in 102 (64.6%) cases,including 42 (42/158,26.6%) EV,27 (27/158,17.1%) HSV Ⅱ,16 (16/158,10.1%) EBV,8 (8/158,5.1%) MP,6(6/158,3.8%) CMV and 3 (3/158,1.9%) HSV Ⅰ.Of these positive samples,10 cases were found with 2～3 pathogens.Conclusion Nested-PCR is of potential value in early pathogenic diagnosis of aseptic meningitis and encephalitis.It can assist laboratory differential diagnosis of central nervous system infections and therefore guide diagnosis and treatment for the diseases.
出处
《中国实用儿科杂志》
CSCD
北大核心
2003年第10期605-608,共4页
Chinese Journal of Practical Pediatrics
基金
北京市自然科学基金"基础性研究实验室项目""北京人类疾病基因诊断实验室"项目资助
湖北科益药业股份有限公司提供部分资助
