A roadmap for rapid molecular differential characterisation of human pathogenic candida species

Al-Laaeiby, A and Abbas, AH ORCID: 0000-0003-2694-730X
(2019) A roadmap for rapid molecular differential characterisation of human pathogenic candida species. EurAsian Journal of BioSciences, 13 (2). pp. 1907-1913.

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The PCR technique is a promising technique used to establish accurate identification methods to gain precise results. The incidence of yeast infections has increased dramatically coinciding with the breakout of HIV infections. A quick and affordable diagnostic procedure is crutial in the quest to tackle such infections. In this study, a detailed diagnostic framework was designed to help in the rapid identification of pathogenic Candida species. A sum of 17 Candida species were involved in this analysis and a common conserved region of rRNA was detected to design primer pairs for in-silico PCR. Restriction Fragment Length Polymorphism (RFLP) was used for species identification. One universal primer was designed to amplify the conserved region in all species, while two others were specific to particular species. Results showed that the PCR-RFLP technique (by using the universal primer No.1) identified 10 species Clavispora lusitaniae, C. Cyberlindnera, C. asseri by unique restriction enzyme HinfI, C. carpophila and C. quercitrusa using HphI and, C. glabrata, Wickerhamomyces anomalus using PhoI while P. kudriavzevii and C. viswanathii using BglI and BseGI respectively. Moreover, C. albicans PCR was identified by double restriction enzymes StuI and BspOI. The PCR-RFLP analysis has identified 7 species using primer pair No.2. These species are C. parapsilosis and Lodderomyces elongisporus by AatI and AcvI respectively. The Cleavage of PCR products using Bst4CI has identified four species (C. glabrata, C. lusitaniae and C. tropicalis). C. dubliniensis was identified by double restriction enzymes (DraI+ ApoI). Meanwhile, C. orthopsilosis was identified through DraI digestion of virtual PCR product (primer pair No.3). All together, these results showed that PCR-RFLP technique is an efficient and rapid method used to scrutinise accurate results by computational based methods before being applied in the Lab. This study set up for a detailed guideline for an accurate, quick diagnostic procedure without any need for the sequencing step.

Item Type: Article
Divisions: Faculty of Health and Life Sciences
Faculty of Health and Life Sciences > Institute of Infection, Veterinary and Ecological Sciences
Depositing User: Symplectic Admin
Date Deposited: 14 May 2021 08:39
Last Modified: 18 Jan 2023 22:47
Open Access URL: http://www.ejobios.org/article/a-roadmap-for-rapid...
URI: https://livrepository.liverpool.ac.uk/id/eprint/3122673