Abstract:
:Genus Candida covers more than 50 species, half of which can cause infections in humans. Some of the Candida species exhibit drug resistance; therefore, there is an urgent need for rapid and accurate differentiation for rendering appropriate and effective management. Here, we report a new methodology employing real-time polymerase chain reaction (RTPCR) and melting temperature analyses (MTA) procedures. Fungal ribosomal internal transcribed spacer 2 (ITS2) has been confirmed with variable nucleotide sequences, which makes it possible to differentiate one species from another by checking their melting temperature following PCR amplification. The universal primers (panFg) covering entire ITS2 region, from 5.8S to 28S rRNA genes, were designed to differentially identify most Candida species with RTPCR-MTA procedure. Nucleic acids from five genomes of closely related Candida species, which were experimentally spiked into human blood, were extracted and amplified. PCR amplicons were called for melting temperature of Candida albicans (87.49°C), C. glabrata (86.85°C), C. krusei (90.24°C), C. parapsilosis (86.22°C), and C. tropicalis (86.08°C). The melting temperature of each amplicon was consistent and reproducible in three replicate experiments (SD ± 0.04-0.32). The new RTPCR-MTA methodology showed promise in differential diagnosis of closely related Candida species from environmental and clinical samples.
journal_name
Mil Medjournal_title
Military medicineauthors
Zhang B,Izadjoo Mdoi
10.7205/MILMED-D-14-00294subject
Has Abstractpub_date
2015-06-01 00:00:00pages
652-9issue
6eissn
0026-4075issn
1930-613Xjournal_volume
180pub_type
杂志文章abstract::After-action reports on the 2005 hurricane relief efforts pointed to the need for effective and efficient civilian/military operational cooperation. Cited for particular attention was the interface with National Guard units. This article describes an exercise conducted with National Guard units and members of the Cent...
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