Comparison of various DNA extraction methods for diagnosis of tuberculosis using a polymerase chain reaction.

Kim, Ju Ock; Han, Pyo Seong; Hong, Seok Cheol; Lee, Jong Jin; Cho, Hai Jeong; Kim, Sun Young

doi:1993.40.1.43

Tuberc Respir Dis > Volume 40(1); 1993 > Article

Original Article

Tuberculosis and Respiratory Diseases 1993;40(1):43-51.
DOI: https://doi.org/10.4046/trd.1993.40.1.43 Published online February 1, 1993.

Comparison of various DNA extraction methods for diagnosis of tuberculosis using a polymerase chain reaction.

Ju Ock Kim, Pyo Seong Han, Seok Cheol Hong, Jong Jin Lee, Hai Jeong Cho, Sun Young Kim

DePartment of Intemal Medicine, School of Medicine, Chungnam National University, Daejeon, Korea

중합효소연쇄반응을 이용한 결핵의 진단에 있어서 각종 DNA 추출방법의 비교

Abstract

Background
The polymerase chain reaction (PCR) is a very sensitive method for the detecting of mycobacterial DNA. There are many reports revealing the efficacy of PCR for the diagnosis of M tuberculosis, but there are many different methods for DNA extraction from Mycobacterium tuberculosis. Bead beater method is a very usef비 method for DNA extraction from clinical specimens, but its procedures are relatively complicated and time-consuming. 50 we studied other methods for the DNA extraction from Mycobacterium tuberculosis H ₃₇Rv and some clinical specimens (5 smear positive sputa and 5 smear negative CSF) .
Methods
We extracted the mycobacterial DNA with 6 different methods from H37Rv strain and clinical specimens. The methods included SDS-microwave oven method , NaOH lysis method, Triton X.100.Proteinase K method, Lysis buffer method, SDS-proteinase K method and bead beater method. The target DNA was 123bp of IS6110 and was detected by examination of ethidium bromide-stained agarose gels.
Results
Among 6 methods, SDS.proteinase K method, bead beater method, lysis buffer method and triton X -100-proteinase K method were excellent, but SDS-proteinase K method was the best method in the aspect of simplicity and cost-effectiveness.
Conclusion
We suggest that SDS-porteinase K method is a simple and convinient method and might be the best method for the extraction of mycobacterial DNA.

Key Words: Polymerase chain reaction, Tuberculosis