一起聚集性腺病毒暴发疫情的调查及病原学分析
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摘要
目的对一起流感样暴发疫情进行流行病学调查和病原学检测,对毒株进行分子流行病学分析,为呼吸道传染病暴发疫情的处置提供依据。方法对暴发疫情进行流行病学调查,收集病例咽拭子样本,采用RespiFinder 2SMART多病原检测试剂盒进行熔解曲线分析,筛查致病原,并以qPCR法确认。用Hep-2细胞分离病毒,提取毒株核酸进行PCR扩增并进行序列分析,NCBI核酸数据库进行序列比对,确定型别。结果疫情累计发病17例,仅涉及一个班级,班级罹患率43.6%,男女比1.8∶1,疫情持续11d。主要症状为发热(17例,100.0%)、咳嗽(16例,94.1%)等。采集10例病例咽拭子,5份以多病原检测试剂盒进行熔解曲线分析,检出2例为腺病毒(ADV)核酸阳性,其余5份经qPCR检测,3份为ADV核酸阳性。以Hep-2细胞分离5份ADV核酸阳性标本,成功分离4株毒株,对其进行DNA测序,NCBI核酸数据库进行Blast序列比对,结果显示其六邻体保守区核酸与ADV3型核酸同源性为97%。结论该暴发是一起由ADV3型引起的暴发疫情。
Objective To carry out epidemiological investigation and etiological analysis on an influenza-like outbreak;to isolate virus strains and analyze their molecular characteristics;to provide experimental evidences for prevention and control of respiratory infectious disease outbreaks.Methods Epidemiological investigation was carried out for the outbreak.Throat swab samples were collected and analyzed by RespiFinder 2SMART multi pathogen analysis kit to identify pathogen by melting curve analysis,which was confirmed by qPCR method.Hep-2cell line was used for virus isolation.The nucleic acid of virus was extracted and amplified for hexon gene by PCR,followed by sequence analysis and Blast in NCBI nucleotide database for to confirm the subtype.Results A total of 17 cases occurred in this outbreak,which all occurred in only one class,resulting class incidence of 43.6%.The ratio of male patients to female patients was 1.8to 1.The main symptoms were fever(17cases,100.0%)and cough(16cases,94.1%),etc.A total of 5throat swabs of 10 collected samples were analyzed by multiple pathogens analysis kit,among which 2were detected positive for adenovirus(ADV)nucleic acid;the other 5samples were analyzed by qPCR,among which 3were positive for ADV nucleic acid.Hep-2cells were used to isolate 5ADV nucleic acid positive samples,a total of 4isolated stains were obtained.DNA sequencing and Blast analysis showed the DNA of hexon gene of isolated viruses had 97% homology with that of ADV3.Conclusion The outbreak was caused by ADV3.
Objective To carry out epidemiological investigation and etiological analysis on an influenza-like outbreak;to isolate virus strains and analyze their molecular characteristics;to provide experimental evidences for prevention and control of respiratory infectious disease outbreaks.Methods Epidemiological investigation was carried out for the outbreak.Throat swab samples were collected and analyzed by RespiFinder 2SMART multi pathogen analysis kit to identify pathogen by melting curve analysis,which was confirmed by qPCR method.Hep-2cell line was used for virus isolation.The nucleic acid of virus was extracted and amplified for hexon gene by PCR,followed by sequence analysis and Blast in NCBI nucleotide database for to confirm the subtype.Results A total of 17 cases occurred in this outbreak,which all occurred in only one class,resulting class incidence of 43.6%.The ratio of male patients to female patients was 1.8to 1.The main symptoms were fever(17cases,100.0%)and cough(16cases,94.1%),etc.A total of 5throat swabs of 10 collected samples were analyzed by multiple pathogens analysis kit,among which 2were detected positive for adenovirus(ADV)nucleic acid;the other 5samples were analyzed by qPCR,among which 3were positive for ADV nucleic acid.Hep-2cells were used to isolate 5ADV nucleic acid positive samples,a total of 4isolated stains were obtained.DNA sequencing and Blast analysis showed the DNA of hexon gene of isolated viruses had 97% homology with that of ADV3.Conclusion The outbreak was caused by ADV3.
引文
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[9]王鹏,曲章义,张鸿彦,等.人腺病毒六邻体蛋白保守区抗原性分析[J].国际免疫学杂志,2007,30(3):135-138.
[10]袁晓辉,曲章义,吴晓敏,等.人3型腺病毒六邻体蛋白同源建模及其进化轨迹分析[J].高等学校化学学报,2009,30(8):1636-1640.
[11]许文波,崔爱利,史智扬,等.江苏省不明原因轻型呼吸道传染病暴发的病因学研究[J].病毒学报,2005,21(5):325-331.
[2]Casas I,Avellon A,Mosquera M,et al.Molecular identification of adenoviruses in clinical samples by analyzing apartial hexon genomic region[J].J Clin Microbiol,2005,43(12):6176-6182.
[3]Bruzzone MS,Fuentes L,Spencer E.Specific subgroup B adenovirus diagnosis by PCR of the fibre gene[J].J Infect,2000,40(2):154-159.
[4]谢立,杨旭辉,陈恩富.腺病毒感染研究进展[J].浙江预防医学,2015,27(3):262-265.
[5]王瑞琨,商蕾,王迎晨,等.呼吸道感染人腺病毒的分离培养和型别鉴定[J].哈尔滨医科大学学报,2013,(5):385-387,482.
[6]Han G,Niu H,Zhao S,et al.Identification and typing of respiratory adenoviruses in Guangzhou,Southern China using a rapid and simple method[J].Virol Sin,2012,28(2):103-8.
[7]Lai C,Lee C,Lu C,et al.Adenovirus Serotype 3and 7Infection with Acute Respiratory Failure in Children in Taiwan,2010-2011[J].PLOS ONE,2013,8(1):e53614.
[8]Lam WY,Yeung AC,Tang JW,et al.Rapid multiplex nested PCR for detection of respiratory viruses[J].J Clin Microbiol,2007,45(11):3631-3640.
[9]王鹏,曲章义,张鸿彦,等.人腺病毒六邻体蛋白保守区抗原性分析[J].国际免疫学杂志,2007,30(3):135-138.
[10]袁晓辉,曲章义,吴晓敏,等.人3型腺病毒六邻体蛋白同源建模及其进化轨迹分析[J].高等学校化学学报,2009,30(8):1636-1640.
[11]许文波,崔爱利,史智扬,等.江苏省不明原因轻型呼吸道传染病暴发的病因学研究[J].病毒学报,2005,21(5):325-331.