Diversity of Nontuberculous Mycobacteria in Kuwait: Rapid Identification and Differentiation of <b><i>Mycobacterium</i></b> Species by Multiplex PCR, INNO-LiPA Mycobacteria v2 Assay and PCR Sequencing of rDNA

Abstract

Objective: Nontuberculous mycobacteria (NTM) often cause disease that is clinically indistinguishable from tuberculosis. Specific identification is important as treatment varies according to Mycobacterium species causing the infection. This study used multiplex PCR (mPCR) assay for rapid differentiation of mycobacterial growth indicator tube 960 system (MGIT) cultures as Mycobacterium tuberculosis (MTB) or NTM together with INNO LiPA Mycobacteria v2 assay (LiPA) and/or PCR sequencing of rDNA for species-specific identification of selected MTB and all NTM isolates in Kuwait. Materials and Methods: DNA was extracted from MGIT cultures (n = 1,033) grown from 664 pulmonary and 369 extrapulmonary specimens from 1,033 suspected tuberculosis patients. mPCR was performed to differentiate MTB from NTM. LiPA was performed and results were interpreted according to kit instructions. rDNA was amplified and sequenced by using panmycobacterial primers. Results: mPCR identified 979 isolates as MTB, 53 as NTM and 1 isolate as mixed culture. LiPA and/or PCR sequencing confirmed 112 of 979 selected isolates as MTB. Mixed culture contained M. tuberculosis and M. fortuitum. LiPA yielded 12 patterns and identified 10 species/species complexes among 47 NTM, M. kansasii + M. scrofulaceum in one culture and 5 isolates only at genus level. PCR sequencing yielded more specific identification for 22 isolates at the species/subspecies level. Conclusions: mPCR rapidly differentiated MTB from NTM. LiPA identified 44 of 52 NTM isolates at the species/species complex level and 2 mixed cultures. PCR sequencing yielded more specific identification at the species/subspecies level. Rapid differentiation as MTB or NTM by mPCR, followed by species-specific NTM identification by LiPA/PCR sequencing is suitable for the proper management of mycobacterial infections in Kuwait.

First insights into the phylogenetic diversity of Mycobacterium tuberculosis in Kuwait and evaluation of REBA MTB-MDR assay for rapid detection of MDR-TB

“…Despite these measures, previous studies have shown that >80% of M . tuberculosis isolates and >90% of MDR-TB strains are isolated from expatriate TB patients in Kuwait [ 4 , 26 , 33 ]. Fingerprinting of M .…”

“…It is also pertinent to mention here that all expatriates entering Kuwait are screened and residence visa is granted only to individuals with no evidence of active TB disease. Despite these measures, previous studies have shown that >80% of M. tuberculosis isolates and >90% of MDR-TB strains are isolated from expatriate TB patients in Kuwait [4,26,33]. Fingerprinting of M. tuberculosis isolates (n = 256) collected from 256 TB patients over a six-month period (October 2017 to March 2018) was performed in this study.…”

Non-tuberculous mycobacteria: clinical and laboratory characterisation (2009 and 2019)

“…Improvement of laboratory methods to identify these species was significant, and it plays a vital role in this https://doi.org/10.1017/S0950268822000899 Published online by Cambridge University Press rising tendency [6]. In this study, changing the test from a probe assay to a PCR-fluorescence probe allowed the identification of 4 more species/subspecies in 2019 (10 vs 6).…”

Performance Comparison of GeneXpert MTB/RIF and ProbeTec ET Tests for Rapid Molecular Diagnosis of Extrapulmonary Tuberculosis in a Low TB/MDR-TB Incidence Country

“…EPTB accounts for ∼15% of all TB cases worldwide with the maximum prevalence of 24% reported from the WHO Eastern Mediterranean Region [1,7]. The Eastern Mediterranean Region includes Kuwait, an Arabian Gulf country where ∼95% of mycobacterial infections are caused by M. tuberculosis [8]. Kuwait has a low incidence of TB (∼24 cases per 100,000 population), high (44%) prevalence of EPTB, and a low (∼1.1%) incidence of MDR-TB [7,9].…”