PERBANDINGAN EKSTRAKSI DNA Salmonella typhi DARI KULTUR DARAH METODE SPIN COLUMN DAN ALCOHOL BASED
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Kultur Darah, Sodium polyanethole sulfonate, Ekstraksi DNA
Abstract
Penggunaan metode biologi molekular seperti PCR dapat mempercepat diagnosis demam tifoid serta memiliki tingkat sensitivitas dan spesifisitas yang tinggi, namun sampel yang berasal dari kultur darah mengandung Sodium polyanethole sulfonate (SPS) yang merupakan inhibitor bagi proses PCR. Proses eliminasi SPS terdapat pada tahap ekstraksi DNA sehingga perlu dipilih metode yang tepat untuk mengekstraksi sampel kultur darah agar proses PCR berhasil. Penelitian ini bertujuan untuk mengetahui perbandingan hasil ekstraksi DNA Salmonella typhi dari kultur darah menggunakan metode Spin column dan Alcohol based. Metode penelitian yang digunakan bersifat eksperimental, dengan membandingkan ekstraksi DNA dari dua metode tersebut ditambah dengan pengenceran sampel 10-1, 10-2, 10-3 dan 10-4 untuk meminimalkan potensi SPS. Hasil ekstraksi kemudian di PCR dan divisualisasikan dengan elektroforesis gel agarose serta didukung dengan pemeriksaan bakteriologis isolat s.typhi pada kultur MCA, uji API 20E dan uji serologi. Berdasarkan hasil penelitian, diketahui bahwa metode Alcohol based berhasil dalam mengekstraksi sampel kultur darah dan menghasilkan band elektroforesis yang jelas hingga pengenceran 10-4 namun metode Spin column gagal karena tidak menghasilkan band elektroforesis bahkan hingga pengenceran 10-4, dari hasil tersebut dapat disimpulkan bahwa metode Alcohol based lebih cocok digunakan untuk mengekstraksi sampel kultur darah dibandingkan metode Spin column.
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