Improved real-time PCR for rapid detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis clinical isolates

Torres, Marı́a José; Criado, Antonio; Ruiz, María Antonia Carmona; Llanos, Ana C.; Palomares, J.C.; Aznar, Javier

doi:10.1016/s0732-8893(02)00521-7

Cited by 59 publications

(61 citation statements)

References 19 publications

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“…127 About 95% of all M. TB RMP-resistant clinical isolates harbour specific mutations within a region of the rpoB gene. 128,129 In contrast to RMP, genotypic testing for INH resistance is much more complex and alterations in several genes including katG and inhA have been reported. 129 This technique uses high-cost equipment beyond the reach of most clinical laboratories except at the reference level.…”

Section: High-performance Liquid Chromatography (Hplc)mentioning

confidence: 99%

A systematic review of rapid diagnostic tests for the detection of tuberculosis infection

Dinnes¹,

Deeks²,

Kunst³

et al. 2007

Health Technol Assess

538

361

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Section: High-performance Liquid Chromatography (Hplc)mentioning

confidence: 99%

A systematic review of rapid diagnostic tests for the detection of tuberculosis infection

Dinnes¹,

Deeks²,

Kunst³

et al. 2007

Health Technol Assess

538

361

View full text Add to dashboard Cite

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“…Understanding the genetic events leading to drug resistance in clinical M. tuberculosis isolates, as well as their prevalence in different geographic areas, makes it possible to develop rapid genetic assays for their detection. 10,32,48 Mutations in an 81-bp "core region" of rpoB gene have been found in approximately 95% of rifampin (RIF)-resistant strains. 7,47 In contrast, resistance to isoniazid (INH) is associated with a variety of mutations affecting one or more genes.…”

Section: Introductionmentioning

confidence: 99%

Molecular Analysis of Isoniazid and Rifampin Resistance inMycobacterium tuberculosisIsolates Recovered from Barcelona

Coll

Aragón²,

Alcaide³

et al. 2005

Microbial Drug Resistance

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We studied the presence of mutations in the whole katG gene and specific regions of the oxyR-ahpC and mabAinhA regulatory region in 61 Mycobacterium tuberculosis isoniazid-resistant isolates. An 81-bp region of the rpoB gene was also sequenced in 17 rifampin-resistant strains. Alterations in the katG gene were detected in 55% of the isolates. Mutation in codon 315 was the most prevalent (32%). Strains showed a high level of resistance, and most maintained a substantial catalase-peroxidase activity. Three strains with an isoniazid MIC of Ն32 g/ml lacked catalase-peroxidase activity. Two of them had deletions in the catalytic domain of the KatG protein. One strain with deletion and three strains with mutations in the C-terminal domain showed low-level resistance and conserved the catalase-peroxidase activity. Mutations in the mabA-inhA regulatory region were identified in 32% of the isolates. All had low-level resistance, and the vast majority conserved catalase-peroxidase activity. Seventeen percent of the isoniazid-resistant isolates had no detectable alterations at the studied loci. Resistance to rifampin was associated with mutations in the 81-bp of the rpoB gene in all cases. IS6110 analysis indicated that recent transmission contributed substantially to the emergence of isoniazid-resistant tuberculosis in Barcelona through short transmission chains. A rapid genotypic assay, including the 315-katG codon and the ؊15 nucleotide of the mabA-inhA regulatory region, may cover 62% of isoniazid-resistant strains in Barcelona. In contrast, the targeting of the 81-bp region of rpoB would detect all our rifampin-resistant isolates. 107

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“…Mutations in the specific codons can therefore be exploited to rapidly detect drug resistance (8,9,12,19,22). Resistance to first-line drugs (INH, RIF) is attributed to mutations in inhA and katG for INH and rpoB for RIF (18,19,22,23).…”

mentioning

confidence: 99%

Rapid Diagnosis of Extensively Drug-Resistant Tuberculosis by Use of a Reverse Line Blot Hybridization Assay

et al. 2011

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Drug resistance in tuberculosis (TB) is a matter of grave concern for TB control programs, as there is currently no cure for some extensively drug-resistant (XDR) strains. There is concern that this resistance could transmit, stressing the need for additional control measures, rapid diagnostic methods, and newer drugs for treatment. We developed an in-house assay that can rapidly detect resistance to drugs involved in the definition of XDR-TB directly from smear-positive specimens. Two hundred fifteen phenotypically XDR-TB isolates and 50 pansusceptible isolates were analyzed using a reverse line blot hybridization (RLBH) assay. The assay was also successfully applied to 73 smear-positive clinical specimens. The RLBH assay exhibited good sensitivity for the detection of resistance to isoniazid (99%), rifampin (99%), fluoroquinolones (95.3%), and second-line aminoglycosides (94.8%). The results from application of this assay on direct smear-positive clinical specimens revealed 93% concordance with the phenotypic drug susceptibility test (DST) results for the above-mentioned drugs. The time to accurate DST results was significantly reduced from weeks to 3 days. This molecular assay is a highly accurate tool for screening for XDR-TB, which achieves a substantial reduction in diagnostic delays.

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Improved real-time PCR for rapid detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis clinical isolates

Cited by 59 publications

References 19 publications

A systematic review of rapid diagnostic tests for the detection of tuberculosis infection

A systematic review of rapid diagnostic tests for the detection of tuberculosis infection

Molecular Analysis of Isoniazid and Rifampin Resistance inMycobacterium tuberculosisIsolates Recovered from Barcelona

Rapid Diagnosis of Extensively Drug-Resistant Tuberculosis by Use of a Reverse Line Blot Hybridization Assay

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