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双链PCR与表型分析在从口腔样本中鉴别都柏林念珠菌和白色念珠菌方面的比较。

Comparison of duplex PCR and phenotypic analysis in differentiating Candida dubliniensis from Candida albicans from oral samples.

作者信息

Sampath Asanga, Weerasekera Manjula, Dilhari Ayomi, Gunasekara Chinthika, Bulugahapitiya Uditha, Fernando Neluka, Samaranayake Lakshman

机构信息

Department of Microbiology, Faculty of Medical Sciences, University of Sri Jayewardenepura, Gangodawila, Nugegoda, Sri Lanka.

Diabetes and Endocrinology Unit, Colombo South Teaching Hospital, Kalubowila, Dehiwala, Sri Lanka.

出版信息

AMB Express. 2017 Dec;7(1):141. doi: 10.1186/s13568-017-0435-9. Epub 2017 Jun 27.

Abstract

Candida dubliniensis shares a wide range of phenotypic characteristics with Candida albicans including a common trait called germ tube positivity. Hence, laboratory differentiation of these two species is cumbersome. Duplex PCR analyses for C. albicans and C. dubliniensis was performed directly on DNA extracted from a total of 122 germ tube positive isolates derived from 100 concentrated oral rinse samples from a random cohort of diabetics attending a clinic in Sri Lanka. These results were confirmed by DNA sequencing of internal transcribed spacer (ITS) region of rDNA of the yeasts. Performance efficacy of duplex PCR was then compared with phenotypic identification using a standard battery of phenotypic tests. Of the 122 germ tube positive isolates three were identified by duplex PCR as C. dubliniensis and the remainder as C. albicans. On the contrary, when the standard phenotypic tests, sugar assimilation and chlamydospore formation, were used to differentiate the two species 13 germ tube positive isolates were erroneously identified as C. dubliniensis. Duplex PCR was found to be rapid, sensitive and more specific than phenotypic identification methods in discriminating C. dubliniensis from C. albicans. This is also the first report on the oral carriage of C. dubliniensis in a Sri Lankan population.

摘要

都柏林念珠菌与白色念珠菌具有广泛的表型特征,包括一种称为芽管阳性的共同特性。因此,这两种菌种的实验室鉴别很繁琐。对从斯里兰卡一家诊所随机选取的糖尿病患者的100份浓缩口腔冲洗样本中分离出的总共122株芽管阳性菌株提取的DNA,直接进行白色念珠菌和都柏林念珠菌的双重PCR分析。通过对酵母核糖体DNA的内转录间隔区(ITS)进行DNA测序,证实了这些结果。然后使用一系列标准表型试验,将双重PCR的性能功效与表型鉴定进行比较。在这122株芽管阳性菌株中,有3株通过双重PCR鉴定为都柏林念珠菌,其余鉴定为白色念珠菌。相反,当使用标准表型试验(糖同化和厚垣孢子形成)来区分这两种菌种时,有13株芽管阳性菌株被错误地鉴定为都柏林念珠菌。发现在区分都柏林念珠菌和白色念珠菌方面,双重PCR比表型鉴定方法更快速、灵敏且更具特异性。这也是关于都柏林念珠菌在斯里兰卡人群口腔中携带情况的首次报告。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/395e/5487313/b2c948799a42/13568_2017_435_Fig1_HTML.jpg

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