[Application of PCR-SSCP technique in detection of rpoB gene mutation in rifampin-resistant Mycobacterium tuberculosis]

Zhonghua Jie He He Hu Xi Za Zhi. 1996 Dec;19(6):338-41.
[Article in Chinese]

Abstract

Objective: To evaluate rpoB gene mutation in rifampin-resistant Mycobacterium tuberculosis and its relationship with rifampin resistance.

Methods: Forty clinical isolates of Mycobacterium tuberculosis were analyzed by PCR-SSCP technique, with H37Rv reference strains as control group.

Results: The sensitivity of amplication products of 411bp and 258bp were found to be 5 pg/microliters, 500 organisms per milliliter and 1 pg/microliter, 500 organisms per milliliter respectively. rpoB gene belongs to genus specificity. Characteristics of SSCP graph of 258bp fragment: Ten sensitive strains were the same as H37 Rv. Thirty strains of rifampin-resistant or multidrug resistance, including rifampin, were different from H37Rv except for three strains. Positive rate was 90%, while specificity 100%.

Conclusions: Results showed that PCR-SSCP technique could detect rPOB gene mutation, which might associate with rifampin resistance and be helpful to rapid detection and research of rifampin-resistant Mycobacterium tuberculosis.

Publication types

  • English Abstract

MeSH terms

  • Antibiotics, Antitubercular / pharmacology
  • Drug Resistance, Microbial / genetics
  • Genes, Bacterial*
  • Humans
  • Mycobacterium tuberculosis / genetics*
  • Mycobacterium tuberculosis / isolation & purification
  • Point Mutation*
  • Polymerase Chain Reaction
  • Polymorphism, Single-Stranded Conformational
  • RNA Polymerase II / genetics*
  • Rifampin / pharmacology
  • Tuberculosis, Pulmonary / microbiology

Substances

  • Antibiotics, Antitubercular
  • RNA Polymerase II
  • Rifampin