Rapid identification of Candida species in oral rinse solutions by PCR
Rapid identification of Candida species in oral rinse solutions by PCR
复制标题
DOI：
10.1136/jcp.2006.040782
复制
发表时间：
2007-09-01
影响因子：
3.4
通讯作者：
Marinelli, Paolo
中科院分区：
文献类型：
--
作者：
Liguori, Giorgio;Lucariello, Angela;Marinelli, Paolo
目的：为了确定的敏感性和特异性的多重PCR检测参与口腔念珠菌病的主要物种的当代识别，没有提取和纯化的DNA从调查中的样品，并评价这种方法与常规的表型培养identifiation.Methods：收集78口腔冲洗液。采用浓缩漱口液技术进行定量和定性研究。进行念珠菌属的研究和鉴定，并进行常规表型培养鉴定（37 ℃下血清中3小时的芽管试验和糖同化条分析）。直接在口腔冲洗液上用多重PCR分析每个样品。在CHROMagar念珠菌平板上重新传代培养常规表型和PCR鉴定方法之间结果不一致的样本。使用真菌特异性引物ITS 1、ITS 2、CA 3和CA 4。用于其他物种的鉴定结果：多重PCR能正确鉴定所有样品，包括单一种、混合种、阴性样品和常规表型法阴性的阳性样品。这种多重PCR检测提供了一种快速的替代传统的文化为基础的技术，最常见的分离念珠菌物种的鉴定和物种。由于没有提取方法，几个小时内就可以鉴定出10个物种。
Aims: To determine the sensitivity and specificity of a multiplex PCR assay for the contemporary identification of major species involved in oral candidiasis, without extraction and purification of DNA from the samples under investigation; and evaluation of this method in comparison with routine phenotypic culture identification.Methods: 78 oral rinse solutions were collected. The concentrated oral rinse technique was used for a quantitative and qualitative study. Research and identification of Candida spp, with routine phenotypic culture identification ( germ-tube test in serum at 37 C for 3 hours and sugar assimilation strip analysis), were performed. Each sample was analysed with multiplex PCR directly on oral rinse solution. Samples giving discrepant results between routine phenotypic and PCR identification methods were resubcultured on CHROMagar Candida plates. The fungus-specific primers ITS1, ITS2, CA3, and CA4 were used. For the identification of other species ( C kefyr, C famata and C dubliniensis), ITS1F, ITS1K, and ITS2D primers were designed.Results: Multiplex PCR correctly identified all samples, including those with single species, or with mixed species, negative samples and positive samples which appeared to be negative from routine phenotypic methods.Conclusion: This multiplex PCR assay provides a rapid alternative to the conventional culture based technique for the identification and speciation of the most frequently isolated Candida species. The absence of an extraction method made identification of 10 species possible in a few hours.