贵阳地区2013年手足口病非 EV71、非 CVA16型肠道病毒的检测分析

目的：了解贵阳地区引起手足口病（HFMD）的非肠道病毒71型（EV71）、非柯萨奇病毒 A（CVA）16型肠道病毒的病原构成及优势型别，为贵阳地区 HFMD 防治工作提供依据。方法收集2013年4～6月检测为非 EV71、非 CVA16型肠道病毒的 HFMD 患者肛拭子标本共107例。提取病毒核酸，用巢式 RT-PCR 法扩增病毒 VP4区序列，对 PCR 阳性扩增产物进行测序，通过与 GenBank 收录的序列 BLAST 比对确定病毒型别；并对优势型别进行序列和进化分析。结果107例标本中共有100例标本巢式 RT-PCR 检测为阳性（93％），其中100例 PCR 产物测序成功，经 BLAST 比对后，100例标本病毒型别均得到确定：CVA6为46例，CVA10为30例，CVA5为10例，CVA2为6例，CVA3和 CVA4各2例，CVB2和 ECHO16各2例。 CVA6在型别确定的非 EV71、非 CVA16型肠道病毒中占42．99％，为优势肠道病毒型别。结论贵阳地区引起 HFMD 的肠道病毒型别多样，CVA6为非 EV71、非 CVA16型肠道病毒中的优势型别。

Etiology study of hand,foot and mouth disease related non-EV71,non-CoxA16 enteroviruses in Guiyang area during 2013

Objective To explore the prevalence of non-EV71 ,non-CoxA16 enteroviruses strains of hand ,foot and mouth disease(HFMD) in Guiyang ,and analyze the genetic evolution of these non-EV71 ,non-CoxA16 enteroviruses strain ,and to provide a basis for preventing and controlling (HFMD) in Guiyang .Methods A total of 107 HFMD specimens tested positive for entero-viruses and negative for EV71 and CoxA16 were collected from April to June of 2013 .The virus RNA was extracted ,and the VP4 sequence was amplified by nested RT-PCR .The PCR positive products were sequenced and the enteroviruses type was identified by sequence BLAST .The sequence and phylogenetic analysis of the predominant virus type was performed .Results 100 out of 107 (93% )specimens were tested positive by nested RT-PCR ,among which 100 PCR products were successfully sequenced .And the types of the 100 enterovirus strains were identifled by sequence BLAST :46 CoxA6 ,30 CoxA10 ,10 CoxA5 ,6 CoxA2 ,2 CoxA3 and 2 CoxA6 ,2 CoxB2 and 2 ECHO16 .CoxA6 accounted for 42 .99% in typed non-EV71 ,non-CoxA16 enterovimses ,and was the pre-dominant type .Conclusion Hand ,foot and mouth disease in Guiyang was conceded with diverse enteroviruses ,and CoxA6 was the predominant type among non-EV71 ,non-CoxA16 enteroviruses .