4种用于产毒型艰难梭菌核酸检测的粪便基因组提取试剂的性能评价  

Performance evaluation of four stool genomic DNA extract reagents for detection of toxigenic Clostridium difficile nucleic acid

作　　者：杨筠 叶菊连[2] 罗芸[2] 宋小军[2] 吴俊 金大智[2] YANG Jun;YE Ju-lian;LUO Yun;SONG Xiao-jun;WU Jun;JIN Da-zhi(Clinical Laboratory,Affiliated Hospital of Hangzhou Normal University,Hangzhou,Zhejiang 310051,China;不详)

机构地区：[1]杭州师范大学附属医院检验科,浙江杭州310051 [2]浙江省疾病预防控制中心,浙江杭州310051 [3]杭州市临安区疾病预防控制中心,浙江杭州310051

出　　处：《中国卫生检验杂志》2019年第22期2699-2702,共4页Chinese Journal of Health Laboratory Technology

基　　金：浙江省重点研发计划(2015C03048);国家卫生计生委科学研究基金-浙江省医药卫生重大科技计划(WKJ-ZJ-1507);浙江省医药卫生科研基金项目(2017KY571)

摘　　要：目的对4种粪便基因组提取试剂进行比较,评价适用于艰难梭菌临床诊断的试剂盒。方法随机选取100份临床腹泻样本,分别采用QIAGEN、天根、美宁康诚和新百基4种试剂提取粪便DNA,采用本实验室研发的艰难梭菌毒素基因检测方法进行检测。结果4种提取试剂的特异度均较好(>95%),敏感度、特异度、阳性预测值和阴性预测值差异均无统计学意义(P>0.05)。而QIAGEN试剂盒的最低检测限(10^3 cfu/ml)高于其他3种试剂盒。4种试剂盒的变异系数均<5%。美宁康诚试剂盒同时操作10份标本的平均用时(30 min)远少于其他3种(F=483.18,P<0.01)。结论4种提取试剂盒均能用于艰难梭菌分子诊断的样本前处理。但美宁康诚试剂盒提取操作时间短,敏感度和特异度均符合检测要求,结果准确,较适合待检艰难梭菌的临床粪便标本的前处理。Objective To compare four stool DNA extraction reagents and evaluate the ones suitable for the clinical diagnosis of Clostridium difficile. Methods A total of 100 samples of clinical diarrhea were randomly collected. The fecal genomic DNA was extracted by QIAGEN, TIANGEN, MokoGene and BIOBASE, and the genomic DNA was detected by the detection method of C. difficile toxin genes developed previously in our laboratory. Results The sensitivity, specificity, positive predictive value and negative predictive value of the four reagents met the needs of clinical diagnostics. The sensitivity(100%) and negative predictive value(100%) of MokoGene kit were the highest among four kits. However, the limit of detection of QIAGEN kit(103 cfu/ml) was higher than other three kits. The coefficients of variation for four kits were less than 5%. The average time in simultaneous operation of 10 samples using MokoGene kit was 30 minutes, which was significantly shorter than those of other three kits(F=483.18, P<0.01). Conclusion All four fecal genomic extraction kits were suitable to be chosen for pre-treatment of molecular diagnostics of toxigenic C.difficile. However, the MokoGene kit with the short operation time, reliable results and good sensitivity and specificity was better for pre-treatment for the detection of toxigenic C.difficile in clinical specimen.

关 键 词：艰难梭菌 核酸提取 评价 

分 类 号：R446.5[医药卫生—诊断学]