摘要
目的评价PCR-反向点杂交法用于念珠菌菌种鉴定及白念珠菌耐药基因突变检测的应用价值。方法收集念珠菌性阴道炎患者及体检健康妇女分泌物各285份和50份。用生物梅里埃酵母菌鉴定卡进行菌种鉴定,用最低抑菌浓度(MIC)法(郑州安图真菌快速培养鉴定药敏试剂)进行药敏试验;采用PCR-反向点杂交法(深圳亚能念珠菌菌种鉴定及白念珠菌耐药基因突变检测试剂)进行菌种鉴定和耐药基因突变检测;采用PCR及测序方法进行耐药基因的检测。分别以培养鉴定法、MIC法、核酸序列测定法为对比方法,评价PCR-反向点杂交法念珠菌菌种鉴定及白念珠菌耐药基因突变检测的敏感性、特异性及准确性。结果与培养鉴定法相比,PCR-反向点杂交法检测6种念珠菌菌种的敏感性、特异性、阳性预测值、阴性预测值和总符合率分别为95%、96%、96%、98%、97%以上,2种方法检测6种念珠菌(白念珠菌、光滑念珠菌、热带念珠菌、近平滑念珠菌、克柔念珠菌和季也蒙念珠菌)结果的差异无统计学意义(χ~2值分别为0.44、0、0、0、0和0,P均>0.05),一致性较好(Kappa均>0.9)。与MIC法相比,PCR-反向点杂交法检测白念珠菌耐药的敏感性、特异性、阳性预测值、阴性预测值和总符合率分别为98%、88%、98%、88%和96%,2种方法检测结果的差异无统计学意义(χ~2=0.17,P>0.05),一致性较好(Kappa>0.8)。PCR-反向点杂交法与核酸序列测定法相比,对6种白念珠菌耐药基因突变位点的检测结果完全一致。结论PCR-反向点杂交法在念珠菌菌种鉴定及白念珠菌耐药基因突变检测上与培养鉴定法以及核酸序列测定法的一致性高,比传统检测方法更早期更快速,可应用于外阴阴道念珠菌病(VVC)的辅助诊断。
Objective To evaluate the application value of PCR-reverse dot blot hybridization in the identification of Candida and the detection of Candida albicans drug-resistant genes. Methods The vaginal secretion samples from 285 patients with candidal vaginitis and 50 healthy women were collected. The identification of Candida species and their drug susceptibility were detected by the bio Mérieux Yeast identification cards and MIC method（ Zhengzhou Antu kit）,respectively. The identification of Candida species and the mutation of Candida albicans,drug-resistant genes were also detected by the Shenzheng Yaneng test kit（ PCR-reverse dot blot hybridization）. The drug-resistant genes were also identified by PCR and nucleic acid sequencing. Based on the culture identification,MIC method and nucleic acid sequencing as the contrast methods,the sensitivity,specificity and accuracy of PCR-reverse dot blot hybridization in the identification of Candida species and the mutation detection of Candida albicans drug-resistant genes were evaluated.Results Compared with the bio Mérieux Yeast identification method,the sensitivity,specificity,positive predictive value,negative predictive value and total coincidence rate of PCR-reverse dot blot hybridization for detecting six kinds of Candida species,including Candida albicans,Candida glabrata,Candida tropicalis,Candida parapsilosis,Candida krusei and Candida guilliermondii,were above 95%,96%,96%,98% and 97%,respectively. There was no significant difference in detecting six kinds of Candida species between the two methods（ χ^2= 0. 44,0,0,0,0 and 0,respectively,P〉 0.05）,and there was good consistency between them（ Kappa 0. 9）. Compared with the MIC method,the sensitivity,specificity,positive predictive value,negative predictive value and total coincidence rate of PCR-reverse dot blot hybridization for detecting the drug resistance of Candida albicans were 98%,88%,98%,88% and 96%,respectively. There was no significant difference in detecting the drug resistance of Candida albicans between the two methods（ χ^2= 0. 17,P〉 0.05）,and there was good consistency between them（ Kappa〉 0. 8）. The results of PCR-reverse dot blot hybridization in detecting the mutation sites of six kinds of Candida albicans drug-resistant genes were 100% of coincidence with that of the nucleic acid sequencing method. Conclusion The PCR-reverse dot blot hybridization has high consistency with the culture method and the nucleic acid sequencing method in the identification of Candida species and the mutation detection of Candida albicans drug-resistant genes,which is more early and rapid than the traditional detection methods,and may be applied to the auxiliary diagnosis of vulvovaginal candidiasis（ VVC）.
作者
刘秀卿
王革非
李卓成
黄磊
LIU Xiu-qing WANG Ge-fei LI Zhno-cheng HUANG Lei(Department of Microbiology and Immunology, Medical College of Shantou University, Shantou 515041, Guangdong Department of Laboratory Medicine, the Second People＇s Hospital of Shenzhen, Shenzhen 518035, Guangdong , China)
出处
《临床检验杂志》
CAS
CSCD
2017年第7期495-498,共4页
Chinese Journal of Clinical Laboratory Science
