目的:本研究的目的是建立一种环介导等温扩增(LAMP)快速检测外阴阴道念珠菌病(VVC)的方法。
方法:我们开发并验证了一种环介导等温扩增(LAMP)方法,用于检测与VVC相关的最常见的念珠菌物种,包括白色念珠菌,格拉布拉斯,C.热带,和拟态C.我们评估了特异性,灵敏度,阳性预测值(PPV),负预测值(NPV),和Kappa值的LAMP方法检测不同种类的念珠菌,使用常规培养方法和内部转录间隔区(ITS)测序作为金标准,涂片革兰氏染色和实时Rolymerase链反应(PCR)作为对照。
结果:共纳入202例,其中88例为VVC阳性,114例为阴性。在88名阳性患者中,真菌培养和ITS测序结果显示67例(76.14%)与白色念珠菌相关,13例(14.77%),光亮奈瑟菌,5(5.68%)与热带梭菌,和3(3.41%)与其他物种。关于整体检出率,LAMP方法具有灵敏度,特异性,PPV,NPV,Kappa值为90.91%,100%,100%,93.4%,分别为0.919。此外,LAMP对白色念珠菌的特异性为100%,格拉布拉斯,和热带C,灵敏度为94.03%,100%,80%,分别。此外,显微镜评估具有最高的灵敏度,而实时PCR对白色念珠菌的特异性低于LAMP。此外,在检测非白色念珠菌(NAC)物种方面,CROMagar念珠菌不如LAMP。
结论:基于成本效益,快速,和廉价的特性的灯,再加上我们的VVC相关念珠菌检测方法的高灵敏度和特异性,我们为VVC的即时测试(POCT)提供了可能性,特别是在发展中国家和一些资源有限的实验室。
OBJECTIVE: The aim of this study is to establish a loop-mediated isothermal amplification (LAMP) method for the rapid detection of vulvovaginal candidiasis (VVC).
METHODS: We developed and validated a loop-mediated isothermal amplification (LAMP) method for detecting the most common Candida species associated with VVC, including C. albicans, N. glabratus, C. tropicalis, and C. parapsilosis. We evaluated the specificity, sensitivity, positive predictive value (PPV), negative predictive value (NPV), and Kappa value of the LAMP method to detect different Candida species, using the conventional culture method and internal transcribed spacer (ITS) sequencing as gold standards and smear Gram staining and real-time Rolymerase Chain Reaction (PCR) as controls.
RESULTS: A total of 202 cases were enrolled, of which 88 were VVC-positive and 114 were negative. Among the 88 positive patients, the fungal culture and ITS sequencing results showed that 67 cases (76.14%) were associated with C. albicans, 13 (14.77%) with N. glabratus, 5 (5.68%) with C. tropicalis, and 3 (3.41%) with other species. Regarding the overall detection rate, the LAMP method presented sensitivity, specificity, PPV, NPV, and Kappa values of 90.91%, 100%, 100%, 93.4%, and 0.919, respectively. Moreover, the LAMP had a specificity of 100% for C. albicans, N. glabratus, and C. tropicalis, with a sensitivity of 94.03%, 100%, and 80%, respectively. Moreover, the microscopy evaluation had the highest sensitivity, while the real-time PCR was less specific for C. albicans than LAMP. In addition, CHROMagar Candida was inferior to LAMP in detecting non-albicans Candida (NAC) species.
CONCLUSIONS: Based on the cost-effective, rapid, and inexpensive characteristics of LAMP, coupled with the high sensitivity and specificity of our VVC-associated Candida detection method, we provided a possibility for the point-of-care testing (POCT) of VVC, especially in developing countries and some laboratories with limited resources.