VP1论文：基因分型诊断在肠道病毒性中枢神经系统感染中的应用探讨.docVIP

VP1论文：基因分型诊断在肠道病毒性中枢神经系统感染中的应用探讨 【中文摘要】基因分型诊断在肠道病毒性中枢神经系统感染病原学诊断的临床应用价值。方法选取我院2006-06-2009-08期间诊断为病毒性脑炎的住院患儿共50例,均于入院的当天抽取脑脊液。试验先经种属特异性引物扩增,筛选出肠道病毒样本,然后分别用测序VP1及VP2序列的方法进行分型。试验同时设立阳性对照及阴性对照。结果50例临床样本中,共有45例临床样本检测出EV RNA。其中,经VP1序列分型方法可成功扩增分型38例,反应产物为357 bp的片段。经VP2序列分型方法可成功扩增分型34例,扩增产物为584 bp的片段。经BLAST比对结果显示：两者在分型结果上有较高的一致性及互补性。结论①经VP1序列分型方法可以成功检测临床常见肠道病毒中的大多数血清型。②联合VP2序列分型方法较单用VP1序列分型方法,分型率及敏感性更高,值得临床上进一步推广。 【英文摘要】the aims of the report were to disscuse the clinic value of a new genic typing method in diagnoseing Enterovirus central nervous system infectionMethods we selecte 50 children who were diagnosed worrying viral encephalitis in our hospital from 2006.06-2009.08, drawing the CSF at the first day of hospitalization. Firstly,we selected the sample infected EV by a general RT-PCR amplification with a pair of primers designed by the 5扷TR of the EV genome. Secondly, we divide the selected samples into two groups. One group we amplify the VP1 region with a pair of specific primers to aim directly at this region.The other we amplify the central of the VP2 region with a pair of specific primers to aim at the region.Thirdly we sequence and subtype the amplified productions.The last we affirm the consequence of the subtype by using the monovalent antiserum corresponding to the type identified by sequencing essays.We also erect the positive control group and the negative control group in the essay.Results EV RNA were detected from 45 clinic samples and 3 positive control samples. Among the total, we successfully amplify the virus genome and gain a 357 bp GE strap by VP1 method from 38 clinic samples and a 584 bp GE strap by VP2 method from 34 clinic samples. By BLAST system contrasting, there is more concordance and complimentarity in the two methods.Conclusion①We can successfully amplify the most serotype of EV by the classic VP1 method in the clinic.②There is more sensibility and higher rate of typing by the VP1 and VP2 method than only by the classic VP1 method.It is