Tesis

Analisa Filogenetik dan Pola Mutasi Full Gen ERG11 Koloni Candida glabrata yang Resisten terhadap Flukonazol di Laboratorium Parasitologi Klinik FKUI = Phylogenetic Analysis and Full Mutation Pattern of ERG11 Gene of Fluconazole Resistant Candida glabrata Colonies in Clinical Parasitology Laboratory of FKUI.

Latar belakang Candida glabrata merupakan penyebab utama kandidiasis non-albicans dengan tingkat mortalitas tinggi. Mekanisme resistensi ini berkaitan dengan mutasi gen ERG11 yang mengkode enzim lanosterol 14-α-demetilase, target utama flukonazol pada jalur biosintesis ergosterol. Tujuan Menentukan pola mutasi gen ERG11 dan menganalisis hubungan filogenetik koloni Candida glabrata resisten flukonazol yang diperoleh dari Laboratorium Parasitologi Klinik FKUI. Metode Penelitian ini merupakan studi deskriptif dengan menggunakan tiga isolat Candida glabrata, 2 sampel berasal dari sampel klinis yang memenuhi kriteria inklusi dari total 19 sampel Candida glabrata dan satu strain referensi ATCC 66032. Identifikasi dilakukan dengan metode kultur, CHROMagar Candida Plus, dan sistem otomatis VITEK 2. DNA diekstraksi menggunakan metode QIAamp DNA Mini Kit, diukur dengan NanoDrop, dan diamplifikasi dengan PCR. Produk PCR divisualisasi melalui elektroforesis gel agarosa dan disekuensing. Hasil sekuensing dianalisis menggunakan BLAST dan dilakukan konstruksi pohon filogenetik. Hasil Hasil identifikasi menunjukkan bahwa kedua isolat klinis merupakan Candida glabrata dengan tingkat resistensi terhadap flukonazol. Analisis sekuensing menunjukkan adanya beberapa substitusi basa nukleotida pada gen ERG11 . Pohon filogenetik menunjukkan adanya kedekatan genetik antara isolat klinis dengan Candida glabrata strain referensi.
Kata Kunci: Candida glabrata, Filogenetik, Flukonazol, Gen ERG11, Resistensi Antijamur


Background Candida glabrata is the main cause of non-albicans candidiasis with a high mortality rate. This resistance mechanism is associated with mutations in the ERG11 gene, which encodes the lanosterol 14-α-demethylase enzyme, the main target of fluconazole in the ergosterol biosynthesis pathway. Objective To determine the pattern of ERG11 gene mutations and analyze the phylogenetic relationship of fluconazole-resistant Candida glabrata colonies obtained from the Clinical Parasitology Laboratory, Faculty of Medicine, University of Indonesia. Methods This study was descriptive in nature and used three Candida glabrata isolates, of the total 19 Candida glabrata samples, two were derived from clinical specimens that met the inclusion criteria, and one reference strain (ATCC 66032) was included in the analysis. Identification was performed using culture methods, CHROMagar Candida Plus, and the VITEK 2 automated system. DNA was extracted using the QIAamp DNA Mini Kit, measured with NanoDrop, and amplified by PCR. PCR products were visualized by agarose gel electrophoresis and sequenced. Sequencing results were analyzed using BLAST and a phylogenetic tree was constructed. Results The identification results showed that both clinical isolates were Candida glabrata with a level of resistance to fluconazole. Sequencing analysis showed several nucleotide base substitutions in the ERG11 gene. The phylogenetic tree showed genetic proximity between the clinical isolates and the Candida glabrata reference strain.
Keywords: Candida glabrata, Phylogenetics, Fluconazole, ERG11 gene, Antifungal resistance