CAJ | 학술논문

目的对2011年5月11-17日厦门某医院收治的10例无菌性脑炎患者进行病原学鉴定.方法采集10例患者的咽拭子、肛拭子及部分患者的脑脊液,进行总肠道病毒核酸检测.总肠道病毒核酸阳性的样本,分别用肠道病毒A、B、C基因型各自的VP1通用引物扩增,VP1区扩增阳性的标本测序后进行序列分析.同时,用Vero细胞进行病毒分离培养,对培养成功的病毒与原始临床标本进行VP1序列同源性分析,从而对引发本次脑炎的病原进行鉴定.结果 10例脑炎患者中,有7例患者的临床标本总肠道病毒核酸检测阳性.这7例患者的咽拭子和(或)肛拭子标本肠道病毒B基因型VP1通用引物扩增阳性,测序后分析表明均为肠道病毒B基因型埃可病毒30(Echo 30),且与浙江2004年流行毒株的同源性达95.3%～97.1%.其中编号为XM2、XM3、XM4、XM8的咽拭子和XM3、XM6的肛拭子相互间序列同源性为99.4%～100.0%,但与XM1的咽拭子序列存在差异,同源性为92.8%～93.4%.此外,XM1、XM2、XM3、XM4、XM8咽拭子标本Vero细胞分离培养病毒成功,其VP1区部分序列与相应的原始标本同源性大于99.9%.结论导致这次无菌性脑炎的病原体为肠道病毒B基因型Echo 30,并提示可能存在多种不同遗传背景的Echo 30在流行.
목적 대2011년5월11-17일하문모의원수치적10례무균성뇌염환자진행병원학감정.방법 채집10례환자적인식자、항식자급부분환자적뇌척액,진행총장도병독핵산검측.총장도병독핵산양성적양본,분별용장도병독A、B、C기인형각자적VP1통용인물확증,VP1구확증양성적표본측서후진행서렬분석.동시,용Vero세포진행병독분리배양,대배양성공적병독여원시림상표본진행VP1서렬동원성분석,종이대인발본차뇌염적병원진행감정.결과 10례뇌염환자중,유7례환자적림상표본총장도병독핵산검측양성.저7례환자적인식자화(혹)항식자표본장도병독B기인형VP1통용인물확증양성,측서후분석표명균위장도병독B기인형애가병독30(Echo 30),차여절강2004년류행독주적동원성체95.3%～97.1%.기중편호위XM2、XM3、XM4、XM8적인식자화XM3、XM6적항식자상호간서렬동원성위99.4%～100.0%,단여XM1적인식자서렬존재차이,동원성위92.8%～93.4%.차외,XM1、XM2、XM3、XM4、XM8인식자표본Vero세포분리배양병독성공,기VP1구부분서렬여상응적원시표본동원성대우99.9%.결론 도치저차무균성뇌염적병원체위장도병독B기인형Echo 30,병제시가능존재다충불동유전배경적Echo 30재류행.
Objective To identify the etiology of an aseptic encephalitis outbreak (ten cases) in a hospital of Xiamen city from 11 to 17 May, 2011.Methods A total of ten patients′ throat swabs, anal swabs and cerebrospinal fluid were collected and detected by RT-PCR for pan-enterovirus. The samples containing detectable pan-enterovirus were tested by PCR with genotype-specific general primers located in VP1 region of enterovirus genotype A, B and C (HEV-A, B and C). The PCR products of VP1 segment were purified and sequenced, and phylogenetic analysis was performed. Meanwhile, the pathogens in those samples were isolated in Vero cell culture. Homologous analysis of VP1 sequences were carried out for the cultured virus samples and the original clinical samples to identify the outbreak etiology.Results Among the ten cases, seven cases were positive for pan-enterovirus nucleic acid. When tested by genotype-specific PCR, the throat and anal swab samples from those 7 patients were positive with HEV-B VP1 primers. Meanwhile, the HEV-B VP1 segments were sequenced and phylogenetic analyzed, which indicated the seven cases were all infected by enterovirus Echo 30. The sequences from those samples had homology of 95.3%-97.1% with the epidemic strains in Zhejiang, 2004. Out of the seven cases, the sequences of XM2,XM3,XM4,XM8 throat swab samples and XM3,XM6 throat samples showed 99.4%-100.0% homology which were different from the sequence of XM1, and the homology was 92.8%-93.4%. Furthermore, the viruses were isolated using Vero cells from XM1,XM2,XM3,XM4 and XM8 throat swab samples,and the VP1 sequence showed more than 99.9% homology with the original specimens.Conclusion The local outbreak of aseptic encephalitis was caused by Echo 30 of enterovirus genotype B, and the epidemic strains may have different genetic background.