应用GeXP遗传分析系统检测病毒性脑炎患儿脑脊液中的人类鼻病毒

Application of GeXP genetic analysis system for detecting human rhinovirus in cerebrospinal fluid of children patients with viral encephalitis

目的探讨儿童脑炎与鼻病毒的关系以及GeXP多重基因表达遗传分析系统在病毒性脑炎患儿脑脊液人类鼻病毒检测方面的应用价值。方法选择2012年1-12月因发热抽搐等症状就诊于汕头大学医学院第二附属医院儿科重症监护室的患儿脑脊液140份,应用GeXP遗传分析系统对脑脊液标本进行包括人类鼻病毒在内的常见呼吸道病毒筛查,巢式RT-聚合酶链反应（PCR）扩增鼻病毒阳性标本,并对扩增产物进行核酸序列测定和同源性比对。结果 GeXP遗传分析系统共检测出6份鼻病毒阳性标本,经巢式RT-PCR扩增、核酸序列测定和同源性比对,结果为鼻病毒C型4份,与GenBank数据库参考序列同源性为95.8%-98.3%。鼻病毒A型和B型各1份,与参考序列同源性分别为96.0%和99.0%。结论鼻病毒是引起儿童脑炎的病原之一,GeXP遗传分析系统为病毒性脑炎的病原学研究提供快速高通量的检测。

Objective To investigate the relationship between the viral encephalitis in children with human rhinovirus（HRV）and the application value of the GenomeLab Gene Expression Profiler（GeXP）analyzer for the detection of HRV in the cerebrospinal fluid（CSF）of children patients with viral encephalitis.Methods 140 CSF specimens from children patients with convulsions and fever admitted to the pediatric intensive care unit（PICU）of the Second Affiliated Hospital of Medical College of Shantou University from January 2012 to December 2012 were collected and tested for common respiratory viruses including HRV by using GeXP genetic analysis system.The HRV positive specimens were amplified by nested RT-PCR,and the positive PCR products were sequenced.The resultant DNA sequences were aligned with the reference HRV sequence in GenBank.Results Totally 6HRV positive specimens were detected by GeXP genetic analysis system.Among them 4specimens were HRV-C genotypes showing95.8%-98.3%identity with reference sequences.The others were 1specimen of HRV-A and 1specimen of HRVB genotype which show 96.0% and 99.0%identity with reference sequences respectively.Conclusion HRV is one of the pathogens causing viral encephalitis in children;the GeXP genetic analysis system could be used as a rapid and high throughput detection tool for viral encephalitis etiology research.