Tuberc Respir Dis.  2012 Jan;72(1):44-49.

Evaluation of Reverse Hybridization Assay for Detecting Fluoroquinolone and Kanamycin Resistance in Multidrug-Resistance Mycobacterium tuberculosis Clinical Isolates

• Park C
• Sung N
• Hwang S
• Jeon J
• Won Y
• Min J
• Kim CT
• Kang H

Abstract

BACKGROUND
Multidrug-resistant tuberculosis (MDR-TB) is an increasing public health problem and poses a serious threat to global TB control. Fluoroquinolone (FQ) and aminoglycoside (AG) are essential anti-TB drugs for MDR-TB treatment. REBA MTB-FQ(R) and REBA MTB-KM(R) (M&D, Wonju, Korea) were evaluated for rapid detection of FQ and kanamycin (KM) resistance in MDR-TB clinical isolates.
METHODS
M. tuberculosis (n=67) were isolated and cultured from the sputum samples of MDR-TB patients for extracting DNA of the bacilli. Mutations in genes, gyrA and rrs, that have been known to be associated with resistance to FQ and KM were analyzed using both REBA MTB-FQ(R) and REBA MTB-KM(R), respectively. The isolates were also utilized for a conventional phenotypic drug susceptibility test (DST) as the gold standard of FQ and KM resistance. The molecular and phenotypic DST results were compared.
RESULTS
Sensitivity and specificity of REBA MTB-FQ(R) were 77 and 100%, respectively. Positive predictive value and negative predictive value of the assay were 100 and 95%, respectively, for FQ resistance. Sensitivity, specificity, positive predictive value and negative predictive value of REBA MTB-KM(R) for detecting KM resistance were 66%, 94%, 70%, and 95%, respectively.
CONCLUSION
REBA MTB-FQ(R) and REBA MTB-KM(R) evaluated in this study showed excellent specificities as 100 and 94%, respectively. However, sensitivities of the assays were low. It is essential to increase sensitivity of the rapid drug resistance assays for appropriate MDR-TB treatment, suggesting further investigation to detect new or other mutation sites of the associated genes in M. tuberculosis is required.

Keyword

Tuberculosis; Fluoroquinolones; Kanamycin; Drug Resistance, Microbial; Mycobacterium tuberculosis; Mutation

MeSH Terms

Chimera
DNA
Drug Resistance
Drug Resistance, Microbial
Fluoroquinolones
Humans
Kanamycin
Kanamycin Resistance
Mycobacterium
Mycobacterium tuberculosis
Public Health
Sputum
Tuberculosis
Tuberculosis, Multidrug-Resistant
DNA
Fluoroquinolones
Kanamycin

Figure

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  Figure 1 Representative patterns of susceptible and resistant strains using REBA MTB-FQ®. P: positive control (H37Rv); 1: no mutation; 2: no mutation+mutation (D94G); 3: mutation (del WT2); 4: mutation (D94A); N: negative control.

• 

  Figure 2 Representative patterns of susceptible and resistant strains using REBA MTB-KM®. P: positive control (H37Rv); 1: mutation (A1401G); 2: no mutation+mutation (A1401G); 3: no mutation; N: negative control.

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