Detection of Pyrazinamide-Resistant Mycobacteriumtuberculosis by PCR-SSCP of pncA Gene / 결핵
Tuberculosis and Respiratory Diseases
;
: 1178-1187, 1998.
Artigo
em Coreano
| WPRIM
| ID: wpr-173317
ABSTRACT
BACKGROUND:
Recently the incidence of tuberculosis is increasing in many countries and control of the disease is further threatened by the emergence of multi-drug resistant tuberculosis. So rapid detection of drug resistance is very important. Pyrazinamide (PZA) is a first-line chemotherapeutic agent for tuberculosis. Now in Korea, we perform PAase activity test instead of actual pyrazinamide susceptibility test for the detection of PZA resistant M. Tuberculosis. Recently the pncA gene, encoding the PZase of M. Tuberculosis, was completely sequenced. And it was reported that the mutation of pncA gene would be associated with PZA resistance of M. Tuberculosis. Therefore we performed this study to evaluate the possibility for the rapid detection of PZA resistant M. Tuberculosis using RCR-SSCR of pncA gene.METHOD:
44 cultured clinical isolates of M. tuberculosis, BCG Tokyo strain, BCG french strain, and one M.bovis isolate were studied. We used H37Rv as the reference strain, The PZase activity test was done at the reference laboratory of Korean Tuberculosis Institute. DNA was extracted by bead-beater method and 561 bp fragment including pncA gene was amplified by PCR. The PCR product were digested by BstB I enzyme. SSCP was done using MDE gel. Of the 44 strains of M. tuberculosis, 22 strains were PZase-positive and other 22 strains were PZase negative.RESULTS:
Of the 22 PZase positive strains, 18 strains(82%) showed the same mobility compared with that of H37Rv and 4(18%) showed different mobility. Of the PZase negative strains, 19(86%) strains showed the same mobility pattern compared with that H37Rv and 3(14%) showed different mobility. Naturally PZA-resistant BCG-French strain, BCG-Tokyo strain, and one M. bovis isolate showed the same band pattern each other, but their mobility were different from that of H37Rv. The results of PZase activity test and PCR-SSCP of pncA of M tuberculosis were statistically significantly correlated each other(p<0.01).CONCLUSION:
The PCR-SSCP after BstB I restriction of pncA gene of M tuberculosis may be a useful method for the rapid detection of PZA-resistant M. tuberculosis.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Pirazinamida
/
Tuberculose
/
DNA
/
Resistência a Medicamentos
/
Reação em Cadeia da Polimerase
/
Incidência
/
Tuberculose Resistente a Múltiplos Medicamentos
/
Polimorfismo Conformacional de Fita Simples
/
Coreia (Geográfico)
/
Mycobacterium bovis
Tipo de estudo:
Estudo diagnóstico
/
Estudo de incidência
/
Estudo prognóstico
País/Região como assunto:
Ásia
Idioma:
Coreano
Revista:
Tuberculosis and Respiratory Diseases
Ano de publicação:
1998
Tipo de documento:
Artigo
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