摘要目的 探讨杭州市轮状病毒检测阴性的腹泻患儿病原体分布情况.方法 收集2017年3月至2018年6月浙江大学附属杭州市第一人民医院,浙江大学附属儿童医院和杭州市儿童医院儿科门诊605例轮状病毒检测阴性的腹泻患儿粪便样本,回顾分析了常规检测结果并对粪便样本性状采用布里斯托评分.粪便样本采用DNA纯化试剂盒和RNA提取试剂盒提取病原体DNA和/或RNA.病原体DNA和RNA逆转录cDNA作为扩增模板,采用聚合酶链反应(PCR)对7种常见致婴幼儿腹泻病原体进行扩增,扩增产物进行琼脂糖凝胶电泳检测.病原体检出阳性率采用卡方检验进行统计学分析.结果 605例患儿中,男性患儿375例(28±11)个月,女性患儿230例(29±10)个月.大便样本布里斯托评分主要集中在6型(496,82％),其次为7型(85,14％)和5型(24,4％).大便常规检测结果中,隐血阳性97例,占比16％,白细胞阳性170例,占比28％.605例患儿大便样本病原体目的基因检出阳性452例,阳性率74.7％.319例检出单一病原体基因片段,127例检出了两种病原体基因,6例检出3种病原体基因片段.检出最高的为艰难梭菌毒素B,阳性率48.9％(296/605),其次为沙门菌属(20.0％,121/605)和诺如病毒(10.9％,66/605).艰难梭菌毒素A阳性率为1.0％(6/605).男性患儿和女性患儿病原体检出阳性率分别为86.7％(325/375)和86.5％(199/230),(χ2=0.002,P=0.959).结论 杭州地区轮状病毒检测阴性的腹泻患儿以沙门菌属和诺如病毒感染为主.艰难梭菌毒素B阳性率高,但艰难梭菌毒素A阳性率低,可能与婴幼儿胃肠道内艰难梭菌定植相关而非腹泻的病原菌.腹泻病原体检出率无性别差异.

abstractsObjective To investigate the distribution of diarrhea pathogens in infants without rotavirus-detection in Hangzhou. Methods 605 stool samples of children with rotavirus-negative diarrhea were collected from Hangzhou First People's Hospital of Zhejiang University, Children's Hospital of Zhejiang University and Hangzhou Children's Hospital from March 2017 to June 2018. The routine test results were analyzed retrospectively and Bristol score was used for the characteristics of stool samples. DNA and/or RNA were extracted from fecal samples with DNA and RNA extraction kit. The extracted DNA and RNA-reversed cDNA were used as templates. 7 common pathogens DNA and/or RNA were amplified by polymerase chain reaction (PCR). The amplified products were detected by agarose gel electrophoresis. The positive rates of pathogens were analyzed by chi-square test. Results Among 605 children, 375 were male (28±11) months and 230 were female (29±10) months. Bristol score of stool samples was mainly in type 6 (496, 82％), followed by type 7 (85, 14％) and type 5 (24, 4％). Among 605 results 97 cases were occult blood positive (positive rate 16％) and 170 cases were white blood cell positive (positive rate 28％).452 of 605 stool samples were positive for pathogen target genes. The positive rate was 74.7％. 319 cases detected single pathogen gene fragments. 127 cases detected two pathogen genes and 6 cases detected three pathogen gene fragments. The positive rate of Clostridium difficile toxin B (48.9％, 296/605)was the highest than the others, followed by Salmonella (20.0％, 121/605) and Norovirus (10.9％, 66/605). The positive rate of Clostridium difficile toxin A was 1.0％ (6/605). The positive rates of pathogens in male and female children were 86.7％(325 / 375) and 86.5％ (199 / 230) respectively, with (χ2 =0.002, P=0.959). Conclusions Salmonella and Norovirus were the main pathogens in children with diarrhea who were negative for rotavirus detection in Hangzhou. The high positive rate of Clostridium difficile toxin B may be related to the colonization of Clostridium difficile in the gastrointestinal tract of infants rather than the pathogen of diarrhea because of the low positive rate of Clostridium difficile toxin A. There was no gender difference in the detection rate of diarrhea pathogens.