食源性腹泻弯曲菌快速鉴定方法评估及耐药性检测  

Evaluation of rapid identification methods for foodborne diarrhea Campylobacter and detection of drug resistance

作　　者：刘畅[1] 杨静 赵建玉 周倩倩 隋文君[1] 杨会娟[3] 鲁辛辛[1] Liu Chang;Yang Jing;Zhao Jianyu;Zhou Qianqian;Sui Wenjun;Yang Huijuan;Lu Xinxin(Department of Laboratory Medicine,Beijing Tongren Hospital,Capital Medical University,Beijing 100730,China;Department of Laboratory Medicine,Beijing Chao-Yang Hospital,Capital Medical University,Beijing 100020,China;Department of Laboratory Medicine,Beijing Puren Hospital,Beijing 100062,China)

机构地区：[1]首都医科大学附属北京同仁医院检验科,北京100730 [2]首都医科大学附属北京朝阳医院检验科,北京100020 [3]北京市普仁医院检验科,北京100062

出　　处：《中华检验医学杂志》2021年第9期821-826,共6页Chinese Journal of Laboratory Medicine

基　　金：国家科技重大专项(2018ZX10102001);国家重点研发计划(2019YFC1200702)。

摘　　要：目的研究并评估适合临床实验室开展的食源性腹泻弯曲菌检测方法及抗菌药物敏感性试验。方法分为预实验和大样本验证。(1)预实验:收集2017年9月至2018年1月就诊于北京地区某三甲医院食源性腹泻患者粪便标本400份。采用双孔滤膜培养法及改良头孢哌酮-木炭-脱氧胆酸盐(CCD)琼脂培养法在微需氧环境中培养48 h,挑选可疑菌落进行基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)鉴定,同时进行实时荧光定量聚合酶链反应(qPCR)检测空肠弯曲菌及结肠弯曲菌。(2)大样本验证:收集2018年4月至2019年3月就诊于北京不同地区不同级别3家医院肠道门诊食源性腹泻患者粪便标本2 062份,进行qPCR检测及培养。阳性菌株按照美国临床与实验室标准化协会及美国国家肠道细菌耐药监测中心推荐的纸片扩散法及琼脂稀释法进行药物敏感性试验并进行结果判读。采用卡方检验比较3种检测方法的检测结果及2种抗菌药物敏感性试验的一致性。结果预实验中qPCR、双孔滤膜培养法及改良CCD琼脂培养法检测弯曲菌(空肠弯曲菌/结肠弯曲菌)的检出率分别是9.0%(36/400)、5.0%(20/400)和3.5%(14/400),差异有统计学意义(χ^(2)=11.772,P<0.01)且qPCR阴性,培养法均为阴性。大样本验证中,qPCR的检出率为8.1%(168/2 062),其中空肠弯曲菌7.0%(144/2 062),结肠弯曲菌1.2%(24/2 062)。qPCR阳性标本进行双孔滤膜培养法培养,阳性率为61.9%(104/168),其中空肠弯曲菌为58.3%(84/144),结肠弯曲菌为83.3%(20/24),与预实验检出率差异无统计学意义(P>0.1)。空肠弯曲菌和结肠弯曲菌对环丙沙星耐药率分别为94.0%(94/100)和100.0%(24/24),对红霉素耐药率分别为6.0%(6/100)和33.3%(8/24),两种抗菌药物敏感性试验方法一致性较好(Kappa值>0.75)。结论 qPCR快速、灵敏、操作简单,适合临床实验室常规开展。双孔滤膜培养法检出率显著高于改良CCD琼脂培养法,弯曲菌�Objective To explore and evaluate a appropriate suitable method for detection of Campylobacter and antibiotic sensitivity test for foodborne diarrhea in clinical laboratories.Methods Pre-experiment:a total number of 400 fecal samples of patients with foodborne diarrhea were prospectively collected from the intestinal disease clinic of Beijing Tongren Hospital from September 2017 to January 2018.Double-hole filtration culture method and modified cefoperazone charcoal deoxycholate(CCD)agar culture method were used for fecal culture in micro-aerobic environment for 48 hours,and then suspicious colonies were identified by matrix assisted laser desorption ionization-time of flight(MALDI-TOF)mass spectrometry.Meanwhile,C.jejuni and C.coli were detected by real-time quantitative polymerase chain reaction(qPCR).Large sample verification:2062 fecal samples of patients with foodborne diarrhea in three hospitals of different levels in different areas of Beijing were collected for qPCR detection and culture from April 2018 to March 2019.The antimicrobial sensitivity test(AST)of C.jejuni and C.coli was performed according to the disk diffusion method and agar dilution method recommended by Clinical and Laboratory Standards Institute and National Antimicrobial Resistance Monitoring System for Enteric Bacteria.The results of the three detection methods and the consistency of the two antibiotic sensitivity tests were compared.Results In the pre-experiment,the positive rates of Campylobacter(jejuni/coli)detected of qPCR,double-hole filtration culture and modified CCD agar culture were 9.0%(36/400),5.0%(20/400)and 3.5%(14/400),and the difference was statistically significant(P<0.01).The samples with negative result of qPCR were negative by both culture methods.The total positive rates of Campylobacter detected by qPCR was 8.1%(168/2062)including 7.0%(144/2062)for C.jejuni and 1.2%(24/2062)for C.coli.The samples with positive qPCR results were cultured by double-hole filtration culture method and the positive rate was 61.9%(104/168

关 键 词：弯曲菌 实时聚合酶链反应 培养 药物敏感性试验 纸片扩散法 琼脂稀释法 

分 类 号：R446.5[医药卫生—诊断学]