Direct Detection of Multidrug-Resistant
            <i>Mycobacterium tuberculosis</i>
            in Clinical Specimens in Low- and High-Incidence Countries by Line Probe Assay

Johansen, Işik Somuncu; Lundgren, Bettina; Sosnovskaja, Anaida; Thomsen, Vibeke Østergaard

doi:10.1128/jcm.41.9.4454-4456.2003

Cited by 49 publications

(36 citation statements)

References 14 publications

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“…The most prevalent mutation found was the rpoB S531L in 86% of the Lithuanian isolates. The rpoB S531L mutation was identified in all Danish RIF-resistant clinical specimens in concordance with a previous study [25]. Studies have shown that mutations at codon 531 within the rpoB gene are the most frequent worldwide [10].…”

Section: Discussionsupporting

confidence: 71%

Multidrug-resistant tuberculosis: rapid detection of resistance to rifampin and high or low levels of isoniazid in clinical specimens and isolates

Vijdea

Stegger

Sosnovskaja

et al. 2008

Eur J Clin Microbiol Infect Dis

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The aim of the present study was to evaluate a new improved multiplex polymerase chain reaction (PCR) hybridisation assay to detect multidrug-resistant tuberculosis. The assay, developed to detect rifampin (rpoB) and isoniazid (katG) gene mutations causing Mycobacterium tuberculosis resistance, was recently extended to include inhA gene mutations that code for low-level isoniazid resistance. Interpretable results were obtained in 115 isolates and in all smear-positive clinical specimens. Rifampin resistance was correctly identified in all specimens and in 20 of 21 (95%) multidrug-resistant isolates compared to BACTEC 460TB. Isoniazid resistance correlated in 18 of 22 (82%) specimens, in 31 of 31 (100%) high-level and 24 of 28 (86%) low-level isoniazid-resistant isolates. The assay was rapid, easy to perform and directly applicable in smear-positive specimens. We predict that the assay may be a useful tool to combat and prevent new cases of multi-and extensively drug-resistant tuberculosis.

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Section: Discussionsupporting

confidence: 71%

Multidrug-resistant tuberculosis: rapid detection of resistance to rifampin and high or low levels of isoniazid in clinical specimens and isolates

Vijdea

Stegger

Sosnovskaja

et al. 2008

Eur J Clin Microbiol Infect Dis

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“…The line probe assays (LPA) GenoType MTBDRplus (Hain Lifescience, Nehren, Germany) and INNO-LiPA Rif.TB (InnoGenetics, Ghent, Belgium) were carried out as previously described (7)(8)(9). With these kits, PCR of rpoB is followed by hybridization of mutants to a membrane strip.…”

Section: P Atients With Tuberculosis (Tb) That Harbor Drug-susceptiblementioning

confidence: 99%

Rifampin Heteroresistance in Mycobacterium tuberculosis Cultures as Detected by Phenotypic and Genotypic Drug Susceptibility Test Methods

et al. 2013

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Tuberculosis patients may harbor both drug-susceptible and -resistant bacteria, i.e., heteroresistance. We used mixtures of rifampin-resistant and -susceptible Mycobacterium tuberculosis strains to simulate heteroresistance in patient samples. Molecular tests can be used for earlier discovery of multidrug resistance (MDR), but the sensitivity to detect heteroresistance is unknown. Conventional phenotypic drug susceptibility testing was the most sensitive, whereas two line probe assays and sequencing were unable to detect the clinically important 1% resistant bacteria. Patients with tuberculosis (TB) that harbor drug-susceptible Mycobacterium tuberculosis strains may also have a small proportion of drug-resistant bacteria that develops spontaneously during replication, normally at a rate of 10 Ϫ8 to 10 Ϫ9 mutations/ cell division (1). For rifampin (Rif) resistance, mutations are almost exclusively found in a single gene, rpoB (2). Conventional drug susceptibility testing (DST) aims to determine if 1% or more of the bacterial population in clinical specimens is drug resistant (3, 4). In this study, cultures that contain both susceptible and at least 1% resistant bacteria are defined as heteroresistant. Heteroresistance is thought to be an early stage in the development of drug-resistant TB in a patient. In such cases, failing to detect resistance may lead to insufficient treatment and treatment failure. As a consequence, spread of resistant bacteria may occur in the future (5). The prevalence of heteroresistance is unknown and is presumably dependent on the local resistance epidemiology. Findings of heteroresistance are accidental, and simple methods for the detection are needed (6).In recent years, a number of genotypic methods have become available for rapid detection of mutations that may confer resistance. Molecular tests have been recommended for use worldwide, with the objective of earlier discovery of multidrug resistance (MDR) (http://www.who.int/tb/features_archive/policy _statement.pdf). These assays are important for the global scaling up of detection of MDR-TB. However, little is known of the sensitivity of these methods to detect resistance in heteroresistant specimens. The aim of the present study was to evaluate the ability of different DST methods to detect Rif resistance when heteroresistance is present.Two freeze-dried strains each of the spoligo families Haarlem and Beijing were obtained from the WHO Tropical Disease Research (TDR) TB Strain Bank. The Haarlem strain TB-TDR-063 was susceptible, and TB-TDR-165 was Rif resistant with the rpoB H526Y mutation. The Beijing strain TB-TDR-077 was susceptible, and TB-TDR-068 was Rif resistant with the rpoB S531L mutation. The susceptible strains from both families had wild-type (WT) DNA in rpoB. The strains were subcultured in Dubos with 0.045% Tween 80 (SSI Diagnostika, Hilleroed, Denmark) with 1 mg/ml Rif (BD, Franklin Lakes, NJ) diluted in water for the resistant strains. After 2 weeks of incubation at 37°C, the bacterial concentrations in liquid medi...

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“…No point mutation in the rpoB gene was detected by sequencing, and thus this phenotype must be due to another, as yet undescribed cause (10,29,31). The predominant mutation detected in the rpoB gene is S531L, the one most frequently detected in European and Asian countries (2,11,13,16,26).…”

mentioning

confidence: 99%

“…The assay was used with appropriate controls, as described in previous evaluations (13,30), namely, inclusion of a negative control containing no mycobacterial DNA (distilled sterile water) and a positive control containing DNA from an M. tuberculosis H37Rv (ATCC 27294) MGIT culture. An initial screening of the kit with nontuberculosis mycobacterial DNA samples (30) was also carried out.…”

mentioning

confidence: 99%

Direct Application of the INNO-LiPA Rif.TB Line-Probe Assay for Rapid Identification of Mycobacterium tuberculosis Complex Strains and Detection of Rifampin Resistance in 360 Smear-Positive Respiratory Specimens from an Area of High Incidence of Multidrug-Resistant Tuberculosis

et al. 2005

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The INNO-LiPA Rif.TB assay for the identification of Mycobacterium tuberculosis complex strains and the detection of rifampin (RIF) resistance has been evaluated with 360 smear-positive respiratory specimens from an area of high incidence of multidrug-resistant tuberculosis (MDR-TB). The sensitivity when compared to conventional identification/culture methods was 82.2%, and the specificity was 66.7%; the sensitivity and specificity were 100.0% and 96.9%, respectively, for the detection of RIF resistance. This assay has the potential to provide rapid information that is essential for the effective management of MDR-TB.

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Direct Detection of Multidrug-Resistant Mycobacterium tuberculosis in Clinical Specimens in Low- and High-Incidence Countries by Line Probe Assay

Cited by 49 publications

References 14 publications

Multidrug-resistant tuberculosis: rapid detection of resistance to rifampin and high or low levels of isoniazid in clinical specimens and isolates

Multidrug-resistant tuberculosis: rapid detection of resistance to rifampin and high or low levels of isoniazid in clinical specimens and isolates

Rifampin Heteroresistance in Mycobacterium tuberculosis Cultures as Detected by Phenotypic and Genotypic Drug Susceptibility Test Methods

Direct Application of the INNO-LiPA Rif.TB Line-Probe Assay for Rapid Identification of Mycobacterium tuberculosis Complex Strains and Detection of Rifampin Resistance in 360 Smear-Positive Respiratory Specimens from an Area of High Incidence of Multidrug-Resistant Tuberculosis

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