Evaluation of the Anyplex™ II MTB/MDR/XDR Kit For Rapid Detection of Drug Resistance in Mycobacterium Tuberculosis Complex Strains By Real-Time PCR
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https://doi.org/10.5281/zenodo.10049082Keywords:
Agar proportion susceptibility method, Extensively Drug Resistance (XDR), Multidrug Resistance (MDR), M. tuberculosis complex (MTBC), Real-Time PCRAbstract
Objective: The occurrence of the extensively drug-resistant (XDR) strains by improving resistance to second-line anti-tuberculosis (anti-TB) drugs of multidrug-resistant (MDR) Mycobacterium tuberculosis complex (MTBC) strains brings about one of the most important human health threats. For this reason, our study aimed to investigate the performance of the multiplex real-time PCR assay, which can be directly performed on clinical samples and simultaneously determines MTBC and the mutations in rapid diagnosis of MDR and XDR strains.
Methods: The study enrolled two groups: the consecutive 44 MDR-MTBC isolates and the consecutive 50 ARB-positive sputum samples phenotypically diagnosed as MTBC resistant to at least one first-line anti-TB drug. Phenotypically, after the identification of MTBC, the drug susceptibility testing (DST) of the isolates for the first- and second-line anti-TB drugs was performed using the BACTEC MGIT 960 system and Löwenstein-Jensen agar proportion (LJ-AP) method, respectively. Genotypically, MTBC positivity and MDR and XDR status were simultaneously investigated by multiplex real-time PCR assay using the Anyplex™ II MTB/MDR/XDR Detection kit (Seegene).
Results: The results of the phenotypic and genotypic investigations were very consistent. The sensitivity and specificity of the real-time PCR assay in MDR detection were 80% and 100%, respectively, compared with phenotypic methods. Two (4.5%) of the 44 MDR-MTBK isolates were identified as pre-XDR-MTBK by both phenotypic and genotypic methods. It was striking that one of these two pre-XDR strains was ARB negative. Moreover, 18 (40.9%) of the MDR-MTBK isolates were ARB(-).
Conclusion: In conclusion, multiplex Real-Time PCR assay using the Anyplex™ II MTB/MDR/XDR Detection kit was suggested to be a usable and reliable method for the detection of MTBC, MDR and XDR in several hours compared with routinely used time-consuming phenotypic methods. However, MDR- and XDR-MTBC strains can be isolated from ARB(+) as well as ARB(-) samples, and investigation of all clinical samples using multiplex real-time PCR assay is very costly. Therefore, following the MTBC identification using a specific cost-effective PCR-based method, confirmation of MTBC and detection of drug resistance with the multiplex real-time PCR detection kit used in our study will be more convenient in the MTBC-positive samples.
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