摘要
目的探讨多重PCR检测5种常见肠杆菌目细菌碳青霉烯酶耐药基因方法的建立及应用评价。方法收集临床分离对碳青霉烯类药物耐药的非重复肠杆菌目细菌118株和对碳青霉烯类药物敏感的肠杆菌目细菌9株,以普通PCR检测blaKPC,blaNDM,blaVIM,blaIMP,blaOXA-48-like结果为参考。采用碳青霉烯酶抑制剂增强试验对试验菌株进行碳青霉烯酶表型检测;构建多重PCR检测5种常见碳青霉烯酶耐药基因。对多重PCR、抑制剂增强试验与普通PCR检测肠杆菌目细菌耐药的灵敏度、特异度和准确度进行比较。结果与普通PCR相比,多重PCR灵敏度为88.54%,特异度为100.00%,κ值为0.79;抑制剂增强试验灵敏度为92.71%,特异度为93.55%,κ值为0.81。在CRE的检测中,虽然多重PCR检测的灵敏度稍低于碳青霉烯酶抑制剂增强试验,但其在特异度上更具优势;二者在阳性率检测上比较差异无统计学意义(P>0.05),检测一致性良好(κ=0.63)。结论该研究成功构建多重PCR体系,能快速、准确检测肠杆菌目细菌中5种常见碳青霉烯酶耐药基因,与临床现行的碳青霉烯酶抑制剂增强试验相比,二者对肠杆菌目细菌耐药检测具有良好的一致性,能极大缩短检测时间。
Objective To establish and application evaluation of multiplex PCR for detection five common carbepenemase genes in Enterobacteriales.Methods The results of blaKPC,blaNDM,blaVIM,blaIMP,blaOXA-48-like resistance genes detection from non-repetitive 118 carbapenem-resistant Enterobacteriales strains and 9 carbapenem-sensitive bacteria was regarded as the reference by the common PCR.Carbapenemase inhibitor enhancement test was used to detect the phenotype in Enterobacteriales.Multiplex PCR was established to detect five common resistance genes.The sensitivity,specificity and accuracy of multiplex PCR,carbapenemase inhibitor enhancement test and common PCR were compared for the detection of Enterobacteriales.Results Compared with the common PCR,the sensitivity of the multiple PCR was 88.54%,the specificity was 100.00%,and theκwas 0.79;the sensitivity of carbapenemase inhibitor enhancement test was 92.71%,and the specificity was 93.55%andκwas 0.81.Although the sensitivity of multiplex PCR was slightly lower than that of carbapenemase inhibitor enhancement test assay,the multiplex PCR had more advantages in terms of specificity.There was no statistically significant difference in positive rate(P>0.05)and the agreement was rated as fair to good(κ=0.63)between the multiplex PCR and carbapenemase inhibitor enhancement test assay.Conclusion Herein,multiple PCR is successfully constructed for rapid and accurate detection of 5 carbapenemase genes in Enterobacteriales.It has a good agreement with the current clinical application of carbapenemase inhibitor enhancement test assay to detect the phenotype in Enterobacteriales,and the detection time is decreased remarkably.
作者
杨冰雪
易淼
袁亚玲
黄金珠
夏培雯
党梓军
廖佳佳
罗胜利
夏云
YANG Bingxue;YI Miao;YUAN Yaling;HUANG Jinzhu;XIA Peiwen;DANG Zijun;LIAO Jiajia;LUO Shengli;XIA Yun(Department of Clinical Laboratory,the First Affiliated Hospital of Chongqing Medical University,Chongqing,400042,China;Department of Clinical Laboratory,Chengdu Second People′s Hospital,Chengdu,Sichuan 610000,China)
出处
《国际检验医学杂志》
CAS
2023年第15期1883-1889,共7页
International Journal of Laboratory Medicine
基金
国家自然科学基金项目(81572055)。
