DOI: http://dx.doi.org/10.18203/2320-6012.ijrms20161227

Multiplex polymerase chain reaction identification of Candida species colonized sputum of patients suffering from various respiratory tract disorders in Duhok, Iraq

Ahmed Basheer Mohammed, Jumma Hussein Ali, Samir Khalaf Abdullah

Abstract


Background: Candida species are part of the body normal flora. Under certain conditions, these opportunistic microorganisms may lead to infection. The purpose of this study was to identify Candida species isolated from sputum from patients suffering from respiratory tract disorders.

Methods: A total of 59 sputum samples taken from patients attending Azadi hospital at Duhok province, Kurdistan Region/Iraq. For primary isolation, sputum samples were cultured on sabouraud dextrose agar (SDA). Suspected colonies of Candida isolates were then sub cultured on chromogenic Candida agar for presumptive identification. Genomic DNA extraction was performed using a genomic DNA extraction kit. For rapid identification of Candida spp, specific primers based on the genomic sequence of DNA topoisomerase 11 of C. albicans, C. parapsilosis I, C. parapsilosis II, C. guilliermondi, C. dubliniensis, C. krusei, C. kefyr and C. glabrata, C. tropicalis I, C. tropicalis II, C. lusitaniae were used. The Multiplex PCR products were separated by electrophoresis in 1.5% agarose gel, visualized by staining with ethidium bromide, and photographed.

Results: Three Candida species namely C. albicans, C. glabrata and C. tropicalis were differentiated by their colour produced on Chromogenic Candida agar. PCR with the primer mixes yielded 4 different sized of PCR products corresponding to C. albicans, C. glabrata, C. Keyfer and C. tropicalis II, C. glabrata was the most common species (33.33%), followed by C. albicans (16.66%). The highest rate of isolation of Candida species was between the ages of 36 to 45.

Conclusion: This study concluded that phenotypic characteristics on selective agar medium such as chromogenic Candida agar are useful for presumptive identification of Candiada spp with the support of molecular method such as multiplex PCR.

 


Keywords


Candida species, Multiplex PCR, Sputum, Chromogenic candida agar, Iraq

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