上海市普陀区儿童感染性腹泻病例病原谱及分子特征

Pathogenic spectrum and molecular characteristics of infectious diarrhea among children in Putuo District, Shanghai

  • 摘要:
    目的 了解2018—2023年上海市普陀区儿童感染性腹泻病的病原谱和分子特征,为儿童感染性腹泻防控提供科学依据。
    方法 收集2018年1月—2023年12月上海市普陀区儿童腹泻病监测哨点医院就诊病例的粪便标本,通过分离培养11种细菌和实时荧光聚合酶链反应(PCR)检测5种病毒。使用脉冲场凝胶电泳(PFGE)标准分型法分析部分阳性菌株的分子分型;采用逆转录PCR方法扩增部分诺如病毒阳性标本的聚合酶⁃衣壳蛋白连接区,基因测序后使用生物信息软件进行分析。采用χ2检验或Fisher精确概率法比较2018—2019年和2020—2023年病原体阳性率。
    结果 707例病例中,总阳性率为47.67%,单一细菌阳性率为16.27%,单一病毒阳性率为22.63%,混合阳性率为8.77%。优势病原体为轮状病毒(10.75%)、诺如病毒(10.33%)、肠致病性大肠杆菌(8.06%)、沙门菌(6.36%)、肠集聚性黏附大肠杆菌(5.52%)、弯曲菌(5.23%)。夏秋季以细菌感染为主,冬春季以病毒感染为主。2020—2023年的总阳性率较2018—2019年下降(χ2=5.753,P<0.05)。对37株沙门菌、81株肠致泻性大肠杆菌、19株弯曲菌完成PFGE分子分型分析,分别分为28种、80种和18种带型,带型相似度范围较广。19份诺如病毒经GⅡ组基因序列分析分为5种基因型,主要为GⅡ.Pe⁃GⅡ.4型和GⅡ.P16⁃GⅡ.2型。
    结论 轮状病毒、诺如病毒、肠致泻性大肠埃希菌、沙门菌、弯曲菌为上海市普陀区儿童感染性腹泻的优势病原体,病原谱呈现季节性流行特征,部分病原体的分子特征存在多样性。应在不同季节加强儿童感染性腹泻病原体的分子特征监测,为防控提供实验室依据。

     

    Abstract:
    Objective To investigate the pathogenic spectrum and molecular characteristics of infectious diarrhea among children in Putuo District of Shanghai from 2018 to 2023, and to provide scientific basis for the prevention and control of infectious diarrhea in children.
    Methods Fecal samples from the cases visited sentinel hospitals for children’s diarrheal disease in Putuo District, Shanghai, were collected from January 2018 to December 2023. A total of 11 species of bacteria were isolated and cultured, and 5 species of viruses were detected by real-time fluorescent polymerase chain reaction (PCR). The molecular typing of some positive strains was analyzed by the standard pulsed-field gel electrophoresis (PFGE) method. The polymerase-capsid protein linkage region of some norovirus-positive samples was amplified by reverse transcription PCR, and was sequenced and analyzed by bioinformatics software. The Chi-square test and Fisher’s exact probability test were used to compare the pathogen-positive rates in 2018‒2019 and 2020‒2023.
    Results Of the 707 cases of children with infectious diarrhea, the total positive rate was 47.67%, with a single bacterial positivity rate of 16.27%, a single viral positivity rate of 22.63%, and a mixed positivity rate of 8.77%, respectively. The dominant pathogens were rotavirus (10.75%), norovirus (10.33%), enteropathogenic Escherichia coli (8.06%), Salmonella (6.36%), enteroaggregative Escherichia coli (5.52%), and Campylobacter (5.23%). Bacterial infections were predominant in summer and fall, and viral infections were predominant in winter and spring. The total positive rate decreased in 2020‒2023 compared with that of 2018‒2019 (χ2=5.753,P<0.05). Thirty-seven strains of Salmonella, 81 strains of diarrheagenic Escherichia coliand 19 strains of Campylobacter were completed for the molecular typing analysis by PFGE, which were classified into 28, 80 and 18 banding types, respectively, with a wide range of banding similarity. Nineteen copies of norovirus GⅡ group gene sequences were analyzed and classified into 5 genotypes, which were mainly GⅡ.Pe-GⅡ.4 and GⅡ.P16-GⅡ.2 types.
    Conclusion Rotavirus, norovirus, diarrheagenic Escherichia coliSalmonella, and Campylobacter are the dominant pathogens of infectious diarrhea in children in Putuo District of Shanghai. The pathogen spectrum shows a trend of seasonal epidemic characteristics, with a diversity of molecular characteristics of some pathogens. Surveillance and monitoring on molecular characteristics of the pathogens of infectious diarrhea in children should be strengthened in different seasons, so as to provide a laboratory basis for the prevention and control of infectious diarrhea.

     

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