摘要
目的获得2008年西安地区肠道病毒71型（EV71）流行株，探寻快速高效的检测技术，分析所获两安地Ⅸ流行株与安徽阜阳流行株的区别。方法采集手足口病患儿的疱液、咽部分泌物进行病毒分离和RT—PCR检测；利用免疫荧光技术进行病毒鉴定。结果56份咽部分泌物接种RD细胞和HEp-2细胞，病毒分离阳性率分别为5．4％（3／56）、1．8％（1／56）；56份疱液标本接种RD细胞和HEp-2细胞，病毒分离阳性率分别为12．5％（7／56）、5．4％（3／56）；RD细胞和HEp-2细胞分别于7d和10d出现细胞病变。检测了56例患儿的咽部分泌物和疱液标本，咽部分泌物RT—PCR结果阳性率为21．4％（12／56），疱液标本RT—PCR结果阳性率为33．9％（19／56）。14份出现细胞病变的细胞株通过免疫荧光鉴定9份为EV71感染。获得了2008年西安地区EV71流行株，将核苷酸序列提交NCBIGenbank，获得核苷酸登录号EU812461。结论本流行株可能嗜上皮细胞性弱、与2008年提交的安徽阜阳病毒株EU703812、EU703813、EU703814同源性达97％～98％；RT—PCR技术有快速、高效的优势，足EV71快速诊断的重要手段。
Objective To isolate the prevalent strain of enterovirus 71 （EV71 ） in Xi＇an area in 2008, and compare the concordance of viral isolation, reverse transcription polymerase chain reaction （ RT- PCR） and immunofluorescent technique in detecting EV71, find the fast and effective method for detection, and analyze the differences between the EV71 strains isolated from Xi＇an and Fuyang, Anhui. Method Virus isolation and RT-PCR were carried out on vesicle fluid and throat swab specimens that were collected from the patients with hand-foot-and-mouth disease, RD and HEp-2 cell lines were used for viral isolation. The virus was indentified by using immunofluorcscence technique. Nucleotide sequencing was performed on positive product of RT-PCR, and compared with EV71 isolated from Fuyang in 2008, then submitted to Genbank. Result Among the 56 samples of throat swab inoculated on RD and HEp-2 cells, the positive rates were 5.4% （3/56） and 1.8% （ 1/56）, respectively. Among the 56 samples of vesicle fluid inoculated on RD and HEp-2 cells, the positive rates were 12.5% （7/56）and 5.4% （3/56）, respectively. Cytopathic effect of RD and HEp-2 cells appeared on days 7 and 10, respectively. The positive rates of RT- PCR on throat swab and vesicle fluid samples were 21.4% （12/56） and 33.9% （19/56）, respectively. Cytopathic effect was found in cell culture for 14 cases and immunofluorescence, showed that 9 of them were infected with EV71. The authors obtained the EV71 strain prevalent in Xi＇an during 2008. The nucleotide sequence was submited to the NCBI Genbank and gained the accession number EU812461. Conclusion The EV71 in Xi＇an prevalent during 2008 may have a weaker epithelial tropism. Comparison of the EV71 strain isolated from Xi＇an with EU703812, EU703813 and EU703814 isolated from Fuyang, Anhui showed that the homology was 97% -98%. RT-PCR is an important method for rapid detection of EV71.
出处
《中华儿科杂志》
CAS
CSCD
北大核心
2009年第7期523-526,共4页
Chinese Journal of Pediatrics
关键词
肠道病毒71型
人
手足口病
肠道病毒感染
逆转录聚合酶链反应
Enterovirus 71, human
Hand, foot and mouth disease
Enterovirus infections
Reverse transcriptase polymerase chain reaction （RT-PCR）
