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Comparative Study
. 2013 Sep 5;8(9):e72036.
doi: 10.1371/journal.pone.0072036. eCollection 2013.

Comparative evaluation of GenoType MTBDRplus line probe assay with solid culture method in early diagnosis of multidrug resistant tuberculosis (MDR-TB) at a tertiary care centre in India

Raj N Yadav  1 , Binit K Singh, Surendra K Sharma, Rohini Sharma, Manish Soneja, Vishnubhatla Sreenivas, Vithal P Myneedu, Mahmud Hanif, Ashok Kumar, Kuldeep S Sachdeva, Chinnambedu N Paramasivan, Balasangameshwra Vollepore, Rahul Thakur, Neeraj Raizada, Suresh K Arora, Sanjeev Sinha
Affiliations
Comparative Study

Comparative evaluation of GenoType MTBDRplus line probe assay with solid culture method in early diagnosis of multidrug resistant tuberculosis (MDR-TB) at a tertiary care centre in India

Raj N Yadav et al. PLoS One. .

Abstract

Background: The objectives of the study were to compare the performance of line probe assay (GenoType MTBDRplus) with solid culture method for an early diagnosis of multidrug resistant tuberculosis (MDR-TB), and to study the mutation patterns associated with rpoB, katG and inhA genes at a tertiary care centre in north India.

Methods: In this cross-sectional study, 269 previously treated sputum-smear acid-fast bacilli (AFB) positive MDR-TB suspects were enrolled from January to September 2012 at the All India Institute of Medical Sciences hospital, New Delhi. Line probe assay (LPA) was performed directly on the sputum specimens and the results were compared with that of conventional drug susceptibility testing (DST) on solid media [Lowenstein Jensen (LJ) method].

Results: DST results by LPA and LJ methods were compared in 242 MDR-TB suspects. The LPA detected rifampicin (RIF) resistance in 70 of 71 cases, isoniazid (INH) resistance in 86 of 93 cases, and MDR-TB in 66 of 68 cases as compared to the conventional method. Overall (rifampicin, isoniazid and MDR-TB) concordance of the LPA with the conventional DST was 96%. Sensitivity and specificity were 98% and 99% respectively for detection of RIF resistance; 92% and 99% respectively for detection of INH resistance; 97% and 100% respectively for detection of MDR-TB. Frequencies of katG gene, inhA gene and combined katG and inhA gene mutations conferring all INH resistance were 72/87 (83%), 10/87 (11%) and 5/87 (6%) respectively. The turnaround time of the LPA test was 48 hours.

Conclusion: The LPA test provides an early diagnosis of monoresistance to isoniazid and rifampicin and is highly sensitive and specific for an early diagnosis of MDR-TB. Based on these findings, it is concluded that the LPA test can be useful in early diagnosis of drug resistant TB in high TB burden countries.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Representative patterns of line probe assay (GenoType MTBDR-plus) strip.
Lane 1, susceptible to rifampicin (RIF) and isoniazid (INH); Lane 2, MDR- TB (rpoB S531L mutation andinhA C15T mutation); Lane 3, rifampicin monoresistant (mutation at rpoB530–533 gene region); Lane 4, absence of TUB band; Lan 5, isoniazid monoresistant (katG S315T1 mutation); Lane 6, DNA positive control (sensitive to rifampicin and isoniazid); Lane 7, DNA negative control.
See this image and copyright information in PMC

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