Metode Cepat Ekstraksi DNA Corynebacterium diphtheriae untuk Pemeriksaan PCR | Perpustakaan dan Galeri Kebijakan Kesehatan

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Metode Cepat Ekstraksi DNA Corynebacterium diphtheriae untuk Pemeriksaan PCR

Sunarno - Nama Orang; Nyoman Fitri - Nama Orang; Fauzul Muna - Nama Orang; Amarila Malik - Nama Orang; Anis Karuniawati - Nama Orang; Amin Soebandrio - Nama Orang;

Diagnosis of diphtheria caused by Corynebacterium diphtheriaeshould be done immediately since delay of therapy may cause 20-fold increase rate of death. One method of rapid diagnostic to identify diphtheria is by using polymerase chain reaction (PCR). The fundamental issue of this method depends on the DNA, either its quality or quantity. The simple DNA extraction method, which is using mechanical/physical principles with a little of chemical reagents (such as boiling method and the use of sodium hydroxide (NAOH)), will have some benefits, such as easy to be
performed, low cost, fast, and environmentally friendly.

This study aimed to evaluate effectivity and efficiency of boiling method with NaOH to extract DNA of C. diphtheriae compared to the use of a commercial diagnostic kit for PCR assay. We used C. diphtheriae toxygenic(NCTC 10648) isolates, which are grown in blood agar plates. We then prepared the suspensions of cell/colony in aquadest with several dilutions. Each dilution was extracted using boiling method, NaOH and controlled with the use of a commercial diagnostic kit (QiAmp DNA Minikit). The results were evaluated quantitatively with spectrophotometer and qualitatively with gel electrophoresis.

The results showed that the extracted DNA from boiling method with NaOH has an adequate quality and quantity for PCR assay (up to 9 CFU/uL cell/reaction). Therefore, it can be summarized that boiling method with NaOH is effective and efficient to be applied in PCR assay for C. diphtheriae.

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Belum memasukkan lokasi Buletin Penelitian Kesehatan, 42 (2) : 85-92

A0004445

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Judul Seri: -
No. Panggil: Buletin Penelitian Kesehatan, 42 (2) : 85-92
Penerbit: Jakarta : Sekretariat Balitbangkes., 2014
Deskripsi Fisik: 8p
Bahasa
ISBN/ISSN: 0125-9695
Klasifikasi: -
Tipe Isi: -
Tipe Media: -
Tipe Pembawa: -
Edisi: -
Subjek: dna
POLYMERASE CHAIN REACTION.
CORYNEBACTERIUM DIPHTHERIAE
Info Detail Spesifik: -
Pernyataan Tanggungjawab: -

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