Metode Cepat Ekstraksi DNA Corynebacterium diphtheriae untuk Pemeriksaan PCR | Perpustakaan Badan Kebijakan Pembangunan Kesehatan

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Metode Cepat Ekstraksi DNA Corynebacterium diphtheriae untuk Pemeriksaan PCR

Sunarno - Personal Name; Fauzul Muna - Personal Name; Nyoman Fitri - Personal Name; Amarila Malik - Personal Name; Anis Karuniawati - Personal Name; Amin Soebandrio - Personal Name;

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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Location name is not set Buletin Penelitian Kesehatan, 42 (2) : 85-92

A0004445

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Series Title: -
Call Number: Buletin Penelitian Kesehatan, 42 (2) : 85-92
Publisher: Jakarta : Sekretariat Balitbangkes., 2014
Collation: 8p
Language
ISBN/ISSN: 0125-9695
Classification: -
Content Type: -
Media Type: -
Carrier Type: -
Edition: -
Subject(s): dna
POLYMERASE CHAIN REACTION.
CORYNEBACTERIUM DIPHTHERIAE
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Statement of Responsibility: -

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