Abstract

BackgroundTuberculosis (TB) is a disease of poverty that contributes significantly to ill-health in developing countries. Drug resistant TB is a major challenge to disease control. Early diagnosis and rapid determination of drug sensitivity is of paramount importance in eradication of TB. Although automated liquid culture based methods are available for rapid detection of drug resistance, the high cost of these tests prevent them from being used routinely in low resource settings. This study compares two phenotypic methods, the manual Mycobacteria Growth Indicator Tube (MGIT) and the Nitrate Reductase Assay (NRA) in liquid medium, with the agar proportion method (APM), the gold standard for susceptibility testing of Mycobacterium tuberculosis.MethodologyFourteen day old M. tuberculosis strains (n=373) grown on solid media were used for drug susceptibility testing by APM, NRA and the manual MGIT method. Rifampicin free and rifampicin incorporated (final concentration, 1 μg/ml) media were inoculated with the recommended concentrations of mycobacterial suspensions and incubated at 37°C in 5% CO2. In the APM, the proportion of colonies in the drug containing medium was determined. In the NRA, the colour change in the medium was compared with a standard colour series after day 6 and day 12 of incubation. Growth in the MGIT was detected using the manual MGIT reader from day 2 onwards. The 2 methods were compared with the gold standard, APM to determine sensitivity and specificity and agreement between the methods was calculated using kappa statistics.ResultsThirty one (31) rifampicin resistant isolates were identified. When compared with the APM, the sensitivity of detection of rifampicin resistance was 85% for the NRA and 93% for the manual MGIT and the specificity was 99% and 100% respectively. Both assays, NRA (κ=0.86) and manual MGIT method (κ= 0.94) were in excellent agreement with the APM. The mean turnaround time for manual MGIT method and NRA were 08 days and 10 days respectively.ConclusionThe NRA in liquid medium and manual MGIT are useful alternatives to APM for drug susceptibility testing of M. tuberculosis in low resource settings.

Highlights

  • Tuberculosis (TB) is a disease of poverty that contributes significantly to ill-health in developing countries

  • The objective of this study was to evaluate the suitability of the manual Mycobacteria Growth Indicator Tube and the nitrate reductase assay for the rapid detection of rifampicin resistance in a low resource setting

  • Thirty one rifampicin resistant isolates were identified among the 373 M. tuberculosis strains isolated during the study period March 2008 to September 2010

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Summary

Introduction

Tuberculosis (TB) is a disease of poverty that contributes significantly to ill-health in developing countries. Drug resistant TB is a major challenge to disease control. Automated liquid culture based methods are available for rapid detection of drug resistance, the high cost of these tests prevent them from being used routinely in low resource settings. The population of Sri Lanka is about 20 million and it is considered a low TB prevalence country in the Asian region. In Sri Lanka, the estimated incidence of all forms of tuberculosis in 2009 was 66 per 100,000 population. The drug resistant rate in Sri Lanka is low. It is around 0.2% among new TB patients and 18%–21% among re-treatment cases. The HIV co-infection rate among TB patients is currently estimated to be less than 0.1% [2]

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