Identification of Candida auris and related species by multiplex PCR based on unique GPI protein‐encoding genes

金念珠菌 基因 多重聚合酶链反应 生物 鉴定(生物学) 遗传学 聚合酶链反应 计算生物学 微生物学
作者
María Alvarado,Joaquín Bartolomé Álvarez,Shawn R. Lockhart,Eulogio Valentín,Alba Ruiz-Gaitán,Elena Eraso,Piet W. J. de Groot
出处
期刊:Mycoses [Wiley]
被引量:5
标识
DOI:10.1111/myc.13204
摘要

Background The pathogen Candida auris is rapidly gaining clinical importance because of its resistance to antifungal treatments and its persistence in hospital environments. Early and accurate diagnosis of C. auris infections is crucial, however, the fungus has often been misidentified by commercial systems. Objectives To develop conventional and real-time PCR methods for accurate and rapid identification of C. auris and its discrimination from closely related species by exploiting the uniqueness of certain glycosylphosphatidylinositol-modified protein-encoding genes. Methods Species-specific primers for two unique putative GPI protein-encoding genes per species were designed for C. auris, C. haemulonii, C. pseudohaemulonii, C. duobushaemulonii, C. lusitaniae, and C. albicans. Primers were blind tested for their specificity and efficiency in conventional and real-time multiplex PCR set-up. Results All primers combinations showed excellent species specificity. In multiplex mode, correct identification was aided by different sized amplicons for each species. Efficiency of the C. auris primers was validated using a panel of 155 C. auris isolates, including all known genetically diverse clades. In real-time multiplex PCR, different melting points of the amplicons allowed the distinction of C. auris from four related species. C. auris limit of detection was 5 CFU/reaction with a threshold value of 32. The method was also able to detect C. auris in spiked blood and serum. Conclusions PCR identification based on unique GPI protein-encoding genes allows for accurate and rapid species identification of C. auris and related species without need for expensive equipment when applied in conventional PCR set-up.
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