1、doi:10.3969/j.issn.1006-5709.2023.01.009基金项目:西京医院学科助推计划(XJZT19ML43)第一作者简介:康蓓佩,硕士,主管技师,研究方向:病原学诊断和院感控制工作。E-mail:742770874 通讯作者:文汉丽,主管护师,研究方向:门诊管理和院内感染控制相关研究。E-mail:306028690 ;周磊,博士,副主任医师,研究方向:病原学诊断和院感控制工作。E-mail:iamwolf-snzq 炎症性肠病与肠道息肉感染致泻性大肠埃希菌实验室检测方法分析康蓓佩1,张鹏亮1,郑恬1,刘昊1,付晓蕊1,孔美娟1,周磊1,文汉丽2空军军医大学西京医院
2、1.检验科;2.门诊部,陕西 西安 710032【摘要】目的分析炎症性肠病(inflammatory bowel disease,IBD)患者和肠道息肉患者感染致泻性大肠埃希菌(diarrheagenic Esch-erichia coli,DEC)实验室检测方法,为该类疾病诊断提供实验室依据。方法收集 2019 年 1 月 1 日至 2020 年 12 月 31 日我院消化内科 130 例 IBD 患者(IBD 组)和 79 例肠道息肉患者(息肉组)的大便,常规方法培养分离大肠埃希菌,用 VITEK Compact 2 全自动细菌分析仪和 VITEK MS 全自动快速微生物质谱检测系统鉴定到
3、种,血清学和分子生物学鉴定 DEC 分型。结果大肠埃希菌分离率 IBD 组高于肠道息肉组(P=0.028)。肠道分离大肠埃希菌性别分布 IBD 组与息肉组比较,差异无统计学意义(P=0.216),年龄分布差异有统计学意义(P=0.023)。大肠埃希菌在麦康凯平板分解乳糖能力 IBD 组与息肉组比较,差异无统计学意义(P=0.112)。采用血清学和实时荧光 PCR 方法进行 DEC 分型鉴定,实时荧光 PCR 阳性 14 株,血清学阳性 13 株,血清学和实时荧光PCR 分型同时阳性 4 株。血清学与实时荧光 PCR 法分型鉴定同时阴性 29 例,符合率 55.78%(29/52);两种方法不一
4、致 19 例,不符合率 36.54%(19/52)。以实时荧光 PCR 分型为准,14 株 DEC 中 IBD 组 12 株,息肉组 2 株,DEC 分离率 IBD 组与息肉组比较,差异无统计学意义(P=0.279)。14 株 DEC 均为 EAEC,年龄 1559 岁,呈季节散发性分布。结论IBD 患者肠道大肠埃希菌分离率高于肠道息肉患者,均为 EAEC,感染以中青年为主。DEC 分型鉴定,传统的血清学实验结果不可靠,建议采用分子生物学方法。【关键词】炎症性肠病;致泻性大肠埃希菌;分型方法中图分类号:R574文献标识码:A文章编号:1006-5709(2023)01-0041-05收稿日期:
5、2021-12-05Analysis of laboratory detection method of diarrheagenic Escherichia coli in inflam-matory bowel disease and intestinal polyp infectionKANG Beipei1,ZHANG Pengliang1,ZHENG Tian1,LIU Hao1,FU Xiaorui1,KONG Meijuan1,ZHOU Lei1,WEN Hanli21.Department of Clinical Laboratory;2.Department of Outpat
6、ient,Xijing Hospital of Air Force Military Medical University,Xian 710032,China【Abstract】ObjectiveTo analyze the laboratory detection method of diarrheagenic Escherichia coli(DEC)in pa-tients with inflammatory bowel disease(IBD)and intestinal polyp,and to provide the laboratory basis for the diagnos
7、is of this disease.MethodsStool samples were collected from 130 patients with IBD and 79 patients with intestinal pol-yps in the Department of Gastroenterology of our hospital from Jan.1st,2019 to Dec.31st,2020.Escherichia coli was cultured and isolated by conventional methods,and identified by VITE
8、K Compact 2 automatic bacterial analyzer and VITEK MS,serology and molecular biology identified and typed of DEC.ResultsThe isolation rate of Escherichia coli in the IBD group was higher than that in the intestinal polyp group(P=0.028).There was no significant difference in gender distribution betwe
9、en IBD group and polyp group(P=0.216),but there was significant difference in age distri-bution(P=0.023).There was no difference in lactose decomposition ability between IBD group and polyp group in MAC plate(P=0.112).Serotyping and real-time PCR were used to identify DEC,14 strains were positive by
10、 real-time PCR,13 strains were positive by serology,and 4 strains were positive by serology and real-time PCR.29 cases were negative simultaneously by serotyping and PCR,and the coincidence rate was 55.78%(29/52).19 cases were incon-sistent between the two methods,and the inconsistent rate was 36.54
11、%(19/52).Based on real-time PCR typing,among the 14 DEC strains,there were 12 strains in IBD group and 2 strains in polyp group.The isolation rate of DEC in IBD group and polyp group had no statistical significance(P=0.279).All the 14 DEC strains were EAEC,aged be-14胃肠病学和肝病学杂志2023 年 1 月第 32 卷第 1 期Ch
12、in J Gastroenterol Hepatol,Jan 2023,Vol.32,No.1tween 15 and 59 years old,and showed sporadic distribution in season.ConclusionThe isolation rate of intestinal Escherichia coli in IBD patients was higher than that in intestinal polyp patients,and EAEC was found in all patients.The infection is mainly
13、 distributed in young and middle-aged patients.The traditional serological test results are not reli-able for the identification of DEC,so molecular biological method is recommended.【Key words】Inflammatory bowel disease;Diarrhegenic Escherichia coli;Classification method炎症性肠病(inflammatory bowel dise
14、ase,IBD)是一种慢性炎症性肠道疾病,其发病被认为是由遗传、环境和免疫因素共同所致。IBD 主要包括:溃疡性结肠炎(ulcerative colitis,UC)和克罗恩病(Crohns disease,CD)。肠道息肉是一种突出到肠腔内、不正常的黏膜或黏膜下增生,需经病理学检查后才可判定其病变性质,多以便血及腹痛为主要症状就诊。致泻性大肠埃希菌(diarrheagenic Escherichia coli,DEC)是一类能引起人体以腹泻症状为主的大肠埃希氏菌。根据致病机制不同可分为肠致病性大肠埃希菌(Enteropathogenic Escherichia coli,EPEC)、肠侵袭性大
15、肠埃希菌(Entero-invasive Escherichia coli,EIEC)、肠产毒性大肠埃希菌(Enterotoxigenic Escherichia coli,ETEC)、产志贺毒素大肠埃希菌(Shiga toxin-producing Escherichia coli)、肠出血性大肠埃希菌(Enterohemorrhagic Escherichia coli,EHEC)和 肠 集 聚 性 大 肠 埃 希 菌(Enteroaggregative Escherichia coli,EAEC)1。本文主要分析 IBD 患者和肠道息肉患者感染 DEC 几种实验室检测方法,为临床诊断 D
16、EC 提供有效的科学依据。1材料与方法1.1菌株来源收集 2019 年 1 月 1 日至 2020 年 12月 31 日我院消化内科 130 例 IBD(UC 和 CD)和 79 例肠道息肉住院患者的大便进行培养,分离大肠埃希菌,鉴定 DEC 并进行分型。所采集标本均来自本院,由检验科技师指导完成。纳入标准:(1)患者均由消化内科明确诊断为 UC、CD 或肠道息肉。(2)住院天数 1 d。排除标准:(1)疑似诊断 IBD,但不能明确为 UC或 CD。(2)伴有其他多系统严重疾病。(3)同一患者重复标本。1.2主要仪器和试剂VITEK Compact 2 全自动细菌分析仪(法国生物梅里埃公司);电子比浊仪(法国生物梅里埃公司);VITEK MS 基质辅助激光解吸电离飞行时间质谱(法国生物梅里埃公司);全自动荧光 PCR检测仪 ABI7500(美国应用生物系统公司);二氧化碳培养箱(上海力申科学仪器有限公司);DEC 血清(日本生物研究所,批号:200754)、5 种 DEC 核酸多重实时荧光 PCR 检测试剂盒(北京卓诚惠生生物科技股份有限公司,批号:A2420-50T)。质控用标准菌株
