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目的对2009年北京市西城区某哨点医院手足口病(hand foot and mouth disease,HFMD)重症病例特征进行初步分析,通过病原学分子分型鉴定,全面了解其病原构成,为重症HFMD的防控提供基础。方法采集重症HFMD患者咽拭子标本,提取病毒RNA。用实时荧光RT-PCR检测肠道病毒(EV)核酸,并对肠道病毒71型(EV71)和柯萨奇病毒A16型(CoxA16)进行分型检测;用肠道病毒VP4区分型引物进行半巢式RT-PCR(RT-nPCR),通过测序和序列分析对非EV71、非CoxA16型EV进行分型鉴定。结果 2009年4～12月,共采集HFMD重症标本35例,发病时间主要集中在6～8月,多为3岁以下婴幼儿。通过实时荧光RT-PCR检测,8例阴性,27例EV阳性,其中,13例EV71阳性,2例CoxA16阳性;12例非EV71、非CoxA16型EV经半巢式RT-nPCR检测,均扩增出目的片段,其中10例测序成功,并通过NCBI BLAST序列比对确定病毒型别:CoxA5和CoxA10各2例,CoxA2、CoxA4、CoxA6、CoxB5、Echoll、人鼻病毒各1例。结论重症手足口病发病高峰为6～8月,3岁以下婴幼儿为主要发病人群。2009年该哨点医院重症HFMD病原型别多样,以EV71为优势型别。
Objective To analyze the characteristics of severe foot and mouth disease (HFMD) in a sentinel hospital in Xicheng District of Beijing in 2009 and identify the pathogen by pathogenic molecular typing. Control provides the basis. Methods Throat swab samples from patients with severe HFMD were collected and viral RNA was extracted. The real-time fluorescent RT-PCR was used to detect the enterovirus (EV) nucleic acid, and the typing of EV71 and CoxA16 was detected. The VP4 genotyping primers were used for half Nested RT-PCR (RT-nPCR) was used to identify non-EV71 and non-CoxA16 EVs by sequencing and sequence analysis. Results From April to December 2009, 35 HFMD severe specimens were collected. The onset time was mainly in June to August, mostly infants under 3 years of age. The results of real-time fluorescent RT-PCR showed that 8 cases were negative and 27 cases were EV positive, of which 13 cases were positive for EV71 and 2 cases were positive for CoxA16. Twelve non-EV71 and non-CoxA16 EVs were detected by semi-nested RT-nPCR A total of 10 cases were successfully sequenced. The virus types were identified by NCBI BLAST sequence analysis: 2 cases of CoxA5 and CoxA10, 1 case of CoxA2, CoxA4, CoxA6, CoxB5, Echoll and human rhinovirus. Conclusions The peak incidence of severe hand-foot-mouth disease is from June to August. Infants under 3 years old are the main pathogenetic group. In 2009 the sentinel hospital for severe HFMD pathogenic diversity, with EV71 as the dominant type.