基于毛细管电泳多重PCR方法对小儿呼吸道感染病原分析  

Analysis of respiratory pathogens for children respiratory tract infection by capillary electrophoresisbased multiplex PCR

作　　者：刘宇奇[1] 黄艳智[1] 孙利伟[1] 姜洪波 田玉玲[1] 孟婷玉[1] 马岩[1] 贺岩[1] 赵艳玲[1] 马英伟[1] Liu Yuqi;Huang Yanzhi;Sun Liwei;Jiang Hongbo;Tian Yuling;Meng Tingyu;Ma Yan;He Yan;Zhao Yanling;Ma Yingwei(Center of Pediatric Precision Medicine Research,Changchun Children's Hospital,Changchun 130000,China;Office of the Directer of the Hospital,Changchun Children's Hospital,Changchun 130000,China)

机构地区：[1]长春市儿童医院儿童精准医学研究中心,130000 [2]长春市儿童医院院长办公室,130000

出　　处：《中国小儿急救医学》2019年第10期764-770,共7页Chinese Pediatric Emergency Medicine

摘　　要：目的探讨基于毛细管电泳的多重PCR方法检测小儿呼吸道感染病原效果,为临床及时有效诊断和治疗提供科学依据.方法按照"十二五"国家科技重大专项发热呼吸道症候群的监测方案要求定义病例;标本来源为2017年1月至2018年2月长春市儿童医院住院的呼吸道感染患儿的鼻咽拭子、支气管肺泡灌洗液、痰液等.应用一步法多重PCR扩增,然后毛细管电泳分离扩增产物,Genemapper software软件分析检测结果.检测病原包括甲型流感病毒(InfA)、人腺病毒(HADV)、博卡病毒(Boca)、人鼻病毒(HRV)、新型InfA﹣09H1(InfA﹣09H1)和季节性流感病毒(InfA﹣H3N2)、副流感病毒(HPIV)、人偏肺病毒(HMPV)、乙型流感病毒(InfB)、肺炎支原体(Mp)、肺炎衣原体(CP)、冠状病毒(HCOV)、人呼吸道合胞病毒(HRSV).结果多重PCR﹣毛细管电泳有效检测率为95.71%;呼吸道病原阳性检出率为62.84%,混合感染率9.61%,混合感染主要为InfA和HRSV;阳性检出率前3位分别是InfA、HRSV、Mp.不同年龄组检出的病原不同,HRSV阳性检出率0~3岁组明显增高;不同季节检出病原不同,呼吸道病毒感染高发季节为12月至次年3月.2017年1月至3月流感流行季以InfA﹣09H1为主;2017年11月、12月流感流行季节以InfA﹣H3N2为主;2017年末2018年初长春地区大范围的流感爆发以InfB为主.HRSV均检出自11月至次年3月份长春地区最寒冷季节.不同感染检出病原不同,HRSV是肺炎患儿最主要的病原,InfA﹣03H2、HPIV是急性支气管炎患儿的主要病原,HRV、InfA是上呼吸道感染患儿的主要病原.结论基于毛细管电泳多重PCR方法检测呼吸道感染病原高效、快速、准确,开发适合临床诊断的试剂,将有广阔的应用前景.Objective To investigate the effect of capillary electrophoresis-based multiplex PCR(CEMP)in detecting pathogens for children respiratory tract infection,and to provide scientific basis for clinical diagnosis and treatment rapidly and accurately.Methods The cases were defined according to the national monitoring program of febrile respiratory syndrome during the 12th Five-Year Plan,and the samples were collected from nasopharyngeal swabs,bronchoalveolar lavage fluid and sputum of children with respiratory tract infection hospitalized in Changchun Children's Hospital from January 2017 to February 2018.Multiplex PCR amplification was performed by one-step method,then PCR products were separated by DNA length size with capillary electrophoresis and pathogens were analyzed byu Genemapper softwareM software.Detecting pathogens included Influenza A virus(InfA),Human Adenovirus(HADV),Boca virus(Boca),Human Rhinovirus(HRV),Novel InfA-09Hl(InfA-09Hl)and Seasonal Influenza virus H3N2(InfAH3N2),Parainfluenza virus(HPIV),Human metapneumonia virus(HMPV),Influenza B virus(InfB),Mycoplasma pneumoniae(Mp),Chlamydia pneumoniae(CP),Human Coronavirus(HCOV),Human Respiratory Syncytial virus(HRSV).Results The effective detection rate of the CEMP assay was 95.71%.The positive detection rate of respiratory tract pathogens was 62.84%and the mixed infection rate was 9.61%.The mixed infection was mainly InfA and HRSV.The highest three positive rates were named InfA,HRSV and Mp.The positive rate of HRSV was significantly higher in the 0-3 age group than that in older group.Different pathogens were detected in different age groups,and the high-occurrence season of respiratory tract infection with virus was from December to March of the next year.InfA-09Hl was the main prevalent influenza virus in January,February and March 2017,InfA-H3N2 was the main prevalent influenza virus in November and December 2017,and the outbreak of InfB was happened in Changchun in late 2017 and early 2018.HRSV was detected only in the coldest season in Changchun

关 键 词：毛细管电泳 呼吸道感染 呼吸道病原 多重PCR 

分 类 号：S85[农业科学—兽医学]