儿童粪便金黄色葡萄球菌毒素基因的检测及分析  

Virulence-associated gene detection and analysis of Staphylococcus aureus isolates from pediatric patients′feces

作　　者：艾晓兰 龙燕[1] 梁秉绍 姚淑雯[2] 刘云锋[1] 高飞[1] 麦嘉良 熊志乐 梁珠薇 王洁琳[1] 陈显堂 杨敏[4] 龚四堂[4] 周珍文[1] Ai Xiaolan;Long Yan;Liang Bingshao;Yao Shuwen;Liu Yunfeng;Gao Fei;Mai Jialiang;Xiong Zhile;Liang Zhuwei;Wang Jielin;Chen Xiantang;Yang Min;Gong Sitang;Zhou Zhenwen(Clinical Laboratory of Zhujiang New Town Hospital,Guangzhou Women and Children′s Medical Center,Guangzhou Medical University,Guangzhou 510623,China;Clinical Laboratory of Children′s Hospital,Guangzhou Women and Children′s Medical Center,Guangzhou Medical University,Guangzhou 510120,China;Clinical Laboratory of Zengcheng Hospital,Guangzhou Women and Children′s Medical Center,Guangzhou Medical University,Guangzhou 511300,China;Gastroenterology Department,Guangzhou Women and Children′s Medical Center,Guangzhou Medical University,Guangzhou 510623,China)

机构地区：[1]广州医科大学广州市妇女儿童医疗中心珠江新城院区检验科,广州510623 [2]广州医科大学广州市妇女儿童医疗中心儿童院区检验科,广州510120 [3]广州医科大学广州市妇女儿童医疗中心增城院区检验科,广州511300 [4]广州医科大学广州市妇女儿童医疗中心消化科,广州510623

出　　处：《中华检验医学杂志》2021年第4期291-297,共7页Chinese Journal of Laboratory Medicine

基　　金：国家自然科学基金(82002202);广东省科技厅项目(2016A020215013);广州市科信局项目(201707010010)。

摘　　要：目的探讨广州地区就诊患儿粪便分离金黄色葡萄球菌毒素基因的分布状况和临床诊断及各分子分型的关系。方法收集2018年8至11月广州市妇女儿童医疗中心3所院区就诊儿童2059份非重复粪便标本所分离的412株金黄色葡萄球菌,采集相关临床信息。提取菌株DNA后用PCR的方法检测相关的毒素基因,其中包括葡萄球菌肠毒素(SE)基因(sea、seb、sec、sed、see)以及杀白细胞毒素基因(pvl),采用多位点序列分型(MLST)的方法对菌株进行分子分型,最后采用χ^(²)检验对数据进行比较。结果从所分离的412株金黄色葡萄球菌中检测到256株(256/412,62.1%)至少含有1种肠毒素基因,携带率较高的依次为sec(125/412,30.3%)、seb(98/412,23.8%)和sea(66/412,16.0%),并且金黄色葡萄球菌pvl基因的携带率为18.7%(77/412)。其中分离自胃肠炎患者粪便的金黄色葡萄球菌sea基因检出率(58/319,18.2%)明显高于非胃肠炎组(8/93,8.6%)(χ^(²)=4.912,P=0.027),而在住院肺炎患者粪便中的金黄色葡萄球菌pvl基因检出率(8/21,38.1%)高于非肺炎组(6/47,12.8%)(χ^(²)=4.252,P=0.039)。并且金黄色葡萄球菌的毒素基因与特定的ST型密切相关,82.4%(28/34)的ST6携带sea基因,所有的ST338和ST59均携带seb基因,96%(48/50)的ST45携带sec基因,ST338的pvl基因携带率高达5/5。结论ST6型金黄色葡萄球菌产生的SEA毒素可能与儿童患者胃肠炎诊断呈密切相关。PVL毒力基因在社区获得性肺炎住院患儿金黄色葡萄球菌中的携带率高于非肺炎组,且与CC59克隆群密切相关。Objective To investigate the multilocus sequence typing feature of the virulence-associated genes of Staphylococcus aureus(S.aureus)separated from the clinical specimens of a multi-center cohort children in Guangzhou area.Methods A total number of 412 Staphylococcus aureus strains isolated from 2059 non-repeated fecal specimens of children by three groups′researchers in Guangzhou Women and Children′s Medical Center from August 2018 to November 2018.While collecting specimens,patient clinical information is also properly collected and preserved.After extracting the DNA of the strain,the virulence-associated genes were detected by polymerase chain reaction(PCR),including the staphylococcal enterotoxin(SE)genes(sea,seb,sec,sed,see)and the Panton-Valentine leucocidin-encoding gene(pvl).The multi-locus sequence typing(MLST)method was performed to reveal the MLST feature of these genes and the statistical difference were examined by the theχ2 test.Results Among the 412 isolates of S.aureus,256 strains(256/412,62.1%)contains at least one SE gene.Among the enterotoxin gens,the sec(125/412,30.3%),seb(98/412,23.8%)and sea(66/412,16.0%)genes were the three most prevalent members of SEs.The frequency of pvl gene in Staphylococcus aureus was 18.7%(77/412).Among them,the frequency of Staphylococcus aureus sea gene isolated from patients with gastroenteritis(58/319,18.2%)was significantly higher than that from the non-gastroenteritis group(8/93,8.6%)(χ^(²)=4.912,P=0.027).The frequency of Staphylococcus aureus pvl gene isolated from the patients with pneumonia(8/21,38.1%)was greater than that from the non-pneumonia group(6/47,12.8%)(χ^(²)=4.252,P=0.039).In addition,the virulence-associated gene of S.aureus was closely related to the specific ST type,82.4%(28/34)of ST6 carried sea gene,all ST338 and ST59 carried seb gene,96%(48/50)ST45 carried sec gene,and the pvl gene carrying rate of ST338 was 5/5.Conclusions The SEA toxin produced by ST6 Staphylococcus aureus may be closely related to the diagnosis of gastroenteritis

关 键 词：葡萄球菌 金黄色 细菌毒素类 肠毒素类 杀白细胞毒素类 多位点序列分型 儿童 

分 类 号：R446.5[医药卫生—诊断学]