Detection of Point Mutations in the rpoB gene Related to Drug Susceptibility in Mycobacterium Tuberculosis using an Oligonucleotide Chip / 결핵
Tuberculosis and Respiratory Diseases
;
: 29-41, 2001.
Artículo
en Coreano
| WPRIM
| ID: wpr-29925
ABSTRACT
BACKGROUND:
The appearance of multiple-drug-resistant Mycobacterium tuberculosis strains has been seriously compromising successful control of tuberculosis. Rifampin-resistance, caused by mutations in the rpoB gene, can be indicative of multiple-drug-resistance, and its detection is of great importance. The present study aimed to develop an oligonucleotide chip for accurate and convenient screening of drug-resistance.METHODS:
In order to detect point mutations in the rpoB gene, an oligonucleotide chip was prepared by immobilizing specific probe DNA to a microscopic slide glass by a chemical reaction. The probe DNA that was selected from the 81 bp core region of the rpoB gene was designed to have mutation sites at the center. A total of 17 mutant probes related to rifampin-resistance including 8 rifabutin-sensitive mutant probes were used in this study. For accurate determination, wild type probes were prepared for each mutation position with an equal length, which enabled a direct comparison of the hybridization intensities between the mutant and wild type.RESULTS:
Mycobacterial genomic DNA from clinical samples was tested with the oligonucleotide chip and the results were compared with those of the drug-susceptibility test in addition to sequencing and INNO-LiPA Rif. TB kit test in some cases. Out of 15 samples, the oligonucleotide chip results of 13 samples showed good agreement with the rifabutin-sensitivity results. The two samples with conflicting result also showed a discrepancy between the other tests, suggesting such possibilities as existence of mixed strains and difference in drug-sensitivity. Further verification of these samples in addition to more case studies are required before the final evaluation of the oligonucleotide chip can be made.CONCLUSION:
An oligonucleotide chip was developed for the detection of rpoB gene mutations related to drug-susceptibility. The results to date show the potential for using the oligonucleotide chip for accurate and convenient screening of drug-resistance to provide useful information in antituberculosis drug therapy.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Rifampin
/
Tuberculosis
/
ADN
/
Tamizaje Masivo
/
Mutación Puntual
/
Rifabutina
/
Quimioterapia
/
Vidrio
/
Mycobacterium
/
Mycobacterium tuberculosis
Tipo de estudio:
Estudio diagnóstico
/
Estudio de tamizaje
Idioma:
Coreano
Revista:
Tuberculosis and Respiratory Diseases
Año:
2001
Tipo del documento:
Artículo
Similares
MEDLINE
...
LILACS
LIS