Retno Sasongkowati (2007) Identifikasi Candida Spesies Menggunakan Primer Campuran Spesifik dengan Teknik PCR Multiplex terhadap Target DNA Topoisomerase II. Masters thesis, UNIVERSITAS AIRLANGGA.

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Abstract

Genus Candida consists of species which can cause fungal opportunistic infections. The species are C. a/bicans, C. tropicalis, C. parapsilosis and C. glabrata. Two enzymatic products of Candida spp, namely aspartyl proteinase and phosphorilase, are virulence factors. The enzymatic products can cause non-specific proteolysis in proteins of the host which involves in the protection against infections and enable yeast to penetrate tissue barrier. Laboratory test using direct or culture clinical materials is needed to find Candida spp. Direct test comprise wet and smear specimens. The test can be successful if the amount of Candida spp fungi is big enough to be seen among tissue cells and other microbes in the clinical materials. This research used Candida isolates in standard isolates media namely Sabouraud Dextrose Agar. Then, the isolates were cultivated in CHROMAgar Candida and made into DNA extraction. The DNA extraction was tested with multiplex PCR using specific mixed primer of PsVIc with DNA topoisomerase II as a target. The purposes of this research were to find out if specific mixed primer of PsVIc can be used to identify Candida until species level. From the research using multiplex PCR, 11 isolates showed positive band of 489,89 bp in isolates number 1, 2, 6, 10, 11, and 672,73 bp in isolates number 3 and 13, and 777,78 bp in isolates number 7, 8, 9 and 12. Therefore, it is concluded that specific mixed primer of PsVIc could be used to identify Candida species.

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