Utility of the REBA MTB-rifa® assay for rapid detection of rifampicin resistant Mycobacterium Tuberculosis

BMC Infectious Diseases

Jong Seok Lee¹  Clifton E Barry 3rd³  Laura E Via³  Sangnae Cho²  Cheon Tae Kim⁴  Doosoo Jeon⁴  Soohee Hwang⁴  Myungsun Lee¹  Jiim Lee¹  Hyun-Kyung Kwak¹  Isdore Chola Shamputa³  Eunjin Cho¹ 

[1] Section of Microbiology, International Tuberculosis Research Center, Changwon 475-1, Republic of Korea;Department of Microbiology, Yonsei University College of Medicine, Seoul, Korea;Tuberculosis Research Section, Laboratory of Clinical Infectious Disease, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA;Masan National Hospital, Changwon 475-1, Korea

关键词: rpoB;    REBA MTB-Rifa® assay;    Rifampicin resistance;    Mycobacterium tuberculosis;   

Others  :  1145695
DOI  :  10.1186/1471-2334-13-478

 received in 2013-07-03, accepted in 2013-10-08,  发布年份 2013

【 摘 要 】

Background

Drug-resistant tuberculosis (TB), including resistance to both rifampicin (RIF) and isoniazid (INH) referred to as multidrug-resistant tuberculosis (MDR-TB), has become an increasing global threat in recent years. Effective management of patients infected with MDR-TB strains requires identifying such patients by performing conventional drug-susceptibility testing (DST) on bacteria isolated from sputum, a process that can take up to 2 months. This delay in diagnosis can result in worsening and continued transmission of MDR-TB. Molecular methods that rely upon nucleic acid amplification of specific alleles known to be associated with resistance to specific drugs have been helpful in shortening the time to detect drug resistant TB.

Methods

We investigated the utility of the REBA MTB-Rifa®, a commercially available line probe assay (LPA) for detecting rifampicin (RIF) resistance in the RIF resistance-determining region (RRDR) of the rpoB gene. Altogether, 492 Mycobacterium tuberculosis (M. tuberculosis) clinical isolates and additional 228 smear- and culture-positive sputum samples with confirmed M. tuberculosis were collected from subjects with suspected MDR-TB in South Korea. The results were compared with conventional phenotypic DST and sequencing of the rpoB gene.

Results

A total of 215 of the 492 isolates were resistant to RIF by conventional DST, and of which 92.1% (198/215) were MDR-TB strains. The REBA MTB-Rifa® assay identified RIF resistance in 98.1% (211/215) of these isolates but failed to identify resistance in four phenotypically RIF resistant isolates. These four isolates lacked mutations in the RRDR but three were confirmed to be MDR-TB strains by sequencing. The sensitivity and specificity of this test for clinical isolates was thus 98.1% (211/215) and 100% (277/277), respectively. When applied directly to 228 smear positive sputum samples, the sensitivity and the specificity of REBA MTB-Rifa® assay was 100% (96/96, 132/132), respectively.

Conclusions

These findings support the use of the REBA MTB-Rifa® assay for rapid detection of RIF resistance on clinical isolates and smear positive sputum samples. The results also suggest that RIF resistance is a good surrogate marker of MDR-TB in South Korea and the need to add more probes to other LPAs which can cover newly identified mutations relevant to RIF resistance.

【 授权许可】

   
2013 Cho et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

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