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目的快速检测深圳市2007年一起急性出血性结膜炎爆发流行的病原并分析其基因进化。方法采集8例疑似急性出血性结膜炎患者的眼拭子标本,提取病毒核酸。用三套可能引起急性出血性结膜炎的病原体的特异性引物进行PCR反应,根据扩增产物琼脂糖凝胶电泳所产生的目的条带快速判定病原体类型。将扩增产物测序后,与不同地区、不同时期相应病原体的毒株进行同源性比较和进化树分析。结果在8例疑似急性出血性结膜炎患者中检测出5例柯萨奇病毒A24型变异株阳性,阳性率为62.5%。基因进化分析显示这5株柯萨奇病毒A24型变异株的VP4序列的组内核苷酸同源性为100%,与同时期广东省流行的CA24v毒株的VP4序列亲缘关系最近,核苷酸同源性为100%;与柯萨奇病毒A24型变异株原型株EH24/70的核苷酸同源性为84.5%;与柯萨奇病毒A24原型株Jospeh的核苷酸同源性为76.3%。结论本次急性出血性结膜炎爆发流行的病原体是柯萨奇病毒A24型变异株,毒株可能来源于同时期广东省流行的株型。
Objective To rapidly detect a pathogen of acute hemorrhagic conjunctivitis outbreak in Shenzhen City in 2007 and analyze its gene evolution. Methods Eye swab samples from 8 patients with suspected acute hemorrhagic conjunctivitis were collected and viral nucleic acids were extracted. Three sets of primers specific for pathogens causing acute hemorrhagic conjunctivitis were used to perform PCR. The type of pathogen was rapidly determined based on the band of interest generated by the agarose gel electrophoresis of the amplification product. The amplified products were sequenced and compared with those of the corresponding pathogens in different regions and different periods for homology comparison and phylogenetic tree analysis. Results In the 8 cases of suspected acute hemorrhagic conjunctivitis, 5 strains of Coxsackievirus A24 were detected positive, the positive rate was 62.5%. Phylogenetic analysis showed that the homology of the VP4 sequences of the 5 strains of Coxsackie virus A24 was 100%, which was the closest to the VP4 sequence of the CA24v strain prevailing in Guangdong during the same period. The nucleotide sequence The nucleotide homology was 84.5% with that of Coxsackievirus A24 strain EH24 / 70 and that of Coxsackievirus A24 prototype strain Jospeh was 76.3 %. Conclusions The pathogen of the outbreak of acute hemorrhagic conjunctivitis is Coxsackievirus A24 variant. The strain may originate from the epidemic strain of Guangdong Province in the same period.