期刊
PATHOGENS
卷 11, 期 2, 页码 -出版社
MDPI
DOI: 10.3390/pathogens11020111
关键词
Candida spp; Candida albicans; DNA extraction; Chelex-100; multiplex LAMP
类别
资金
- Korea Health Technology R&D Project, through the Korea Health Industry Development Institute (KHIDI) - Ministry of Health and Welfare, Republic of Korea [HR20C0021]
- National Research Foundation of Korea (NRF) - Korean government (MEST) [2020R1A2C101176511]
To reduce the morbidity and mortality of candidemia patients, a simple and rapid molecular diagnostic method based on nucleic acid extraction is needed. We developed a multiplex Candida Pan/internal control (IC) loop-mediated isothermal amplification (LAMP) assay and a Chelex-100 boiling protocol for DNA extraction. The results showed high sensitivity and specificity, suggesting that this method could be used as a rapid molecular diagnostic test for candidemia.
To reduce the morbidity and mortality of candidemia patients through rapid treatment, the development of a simple, rapid molecular diagnostic method that is based on nucleic acid extraction and is superior to conventional methods for detecting Candida in the blood is necessary. We developed a multiplex Candida Pan/internal control (IC) loop-mediated isothermal amplification (LAMP) assay and a simple DNA extraction boiling protocol using Chelex-100 that could extract yeast DNA in blood within 20 min. The Chelex-100/boiling method for DNA extraction showed comparable efficiency to that of the commercial QIAamp UCP Pathogen Mini Kit using Candida albicans qPCR. In addition, the Candida Pan/IC LAMP assay showed superior sensitivity to that of general Candida Pan and species qPCRs against clinical DNA samples extracted with the QIAamp UCP Pathogen Mini Kit and Chelex-100/boiling method. The Candida Pan/IC LAMP assay followed by Chelex-100/boiling-mediated DNA extraction showed high sensitivity (100%) and specificity (100%) against clinical samples infected with Candida. These results suggest that the Candida Pan/IC LAMP assay could be used as a rapid molecular diagnostic test for candidemia.
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