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目的:为探索手足口病的病原并为该病的防治提供科学依据,对本省徐州地区采集的19份手足口病临床标本进行病原学分离与鉴定。方法:采集的患者咽拭子(6份)和肛拭子(13份)标本,接种Hep-2细胞进行病毒分离,经培养3代,对有明显致细胞病变效应并能连续传代的毒株,用肠道病毒通用引物real-time-RT-PCR方法鉴定确认为肠道病毒后,用肠道病毒分子分型引物(E187/E222)对肠道病毒VP1区进行扩增,将扩增产物测序获得的序列与GeneBank中肠道病毒核苷酸序列进行比对,确定毒株血清型别。结果:19份标本中分离到9株病毒,均为肠道病毒。对其VP1区部分核苷酸序列测定和比较分析后,鉴定出其中有6株为CoxB5,1株为CoxB3,2株为Echo6。结论:虽然肠道病毒71型(EV71)和柯萨奇病毒A16(CoxA16)是引起手足口病暴发流行的主要病原,但引起手足口病的肠道病毒存在多种血清型,CoxB5、CoxB3和Echo6等均能引起手足口病。因此,对手足口病的病原学检测,除EV71和CoxA16外,不能忽视对其他肠道病毒的病原学检测和鉴定。
Objective: To explore the pathogens of hand-foot-mouth disease and provide a scientific basis for the prevention and treatment of the disease, 19 hand-foot-mouth disease clinical specimens collected from Xuzhou area in our province were etiologically isolated and identified. Methods: Throat swabs (6) and anal swabs (13) were collected and inoculated into Hep-2 cells for virus isolation. After three generations of culture, the strains with obvious cytopathic effect and continuous passage , And the enterovirus was identified by real-time-RT-PCR using the universal primer of enterovirus. After the enterovirus VP1 region was amplified by enterovirus molecular typing primer (E187 / E222), the amplified product The sequences obtained by sequencing were aligned with the nucleotide sequences of enterovirus in GeneBank to determine the serotypes of the strains. Results: Nine strains of viruses were isolated from 19 samples, all of which were enteroviruses. After partial nucleotide sequence analysis and comparison of VP1 region, 6 strains were identified as CoxB5, 1 strain as CoxB3 and 2 as Echo6. CONCLUSIONS: Although enterovirus 71 (EV71) and CoxA16 (CoxA16) are the major pathogens that cause HFMD outbreaks, there are several serotypes of enterovirus causing CIMB, CoxB5, CoxB3 and Echo6 and so can cause hand, foot and mouth disease. Therefore, the etiology of foot-mouth disease detection, in addition to EV71 and CoxA16, can not ignore the etiology of other enterovirus detection and identification.