IPICYT, División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica, AC, Camino a la Presa San José No. 2055, Col. Lomas 4, 78216, San Luis Potosí, Mexico.
Braz J Microbiol. 2021 Dec;52(4):1733-1744. doi: 10.1007/s42770-021-00584-2. Epub 2021 Jul 31.
The most common nosocomial fungal infections are caused by several species of Candida, of which Candida glabrata is the second most frequently isolated species from bloodstream infections. C. glabrata displays relatively high minimal inhibitory concentration values (MIC) to the antifungal fluconazole and is associated with high mortality rates. To decrease mortality rates, the appropriate treatment must be administered promptly. C. glabrata contains in its genome several non-identical copies of species-specific sequences. We designed three pairs of C. glabrata-specific primers for endpoint PCR amplification that align to these species-specific sequences and amplify the different copies in the genome. Using these primers, we developed a fast, sensitive, inexpensive, and highly specific PCR-based method to positively detect C. glabrata DNA in a concentration-dependent manner from mixes of purified genomic DNA of several Candida species, as well as from hemocultures and urine clinical samples. This tool can be used for positive identification of C. glabrata in the clinic.
最常见的医院真菌感染是由几种念珠菌引起的,其中光滑念珠菌是血流感染中第二常见的分离菌种。光滑念珠菌对抗真菌药物氟康唑的最小抑菌浓度值(MIC)相对较高,且与高死亡率相关。为了降低死亡率,必须及时给予适当的治疗。光滑念珠菌的基因组中含有几个非同源的种特异性序列的拷贝。我们设计了三对针对端点 PCR 扩增的光滑念珠菌特异性引物,这些引物与这些种特异性序列对齐,并在基因组中扩增不同的拷贝。使用这些引物,我们开发了一种快速、敏感、廉价且高度特异性的基于 PCR 的方法,能够以浓度依赖的方式从几种念珠菌的纯化基因组 DNA 混合物、血培养物和尿液临床样本中,正向检测出依赖浓度的光滑念珠菌 DNA。该工具可用于临床中对光滑念珠菌的阳性鉴定。