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Evaluation of the diagnostic value of loop-mediated isothermal amplification assays targeting three different Mycobacterium tuberculosis genes. | LitMetric

Evaluation of the diagnostic value of loop-mediated isothermal amplification assays targeting three different Mycobacterium tuberculosis genes.

J Microbiol Methods

Disease Prevention and Control Department, Beijing Chest Hospital, Capital Medical University, Beijing Tuberculosis and Thoracic Tumor Research Institute, Beijing 101149,China. Electronic address:

Published: September 2022

AI Article Synopsis

  • - The study aimed to evaluate a loop-mediated isothermal amplification (LAMP) assay for quick and direct diagnosis of tuberculosis from clinical samples.
  • - Three LAMP assays targeting specific Mycobacterium tuberculosis genes were tested, achieving high sensitivity (up to 98.2%) and 100% specificity against non-tuberculosis species in controlled and clinical settings.
  • - LAMP proved to be more sensitive than traditional smear microscopy and was almost as sensitive as quantitative real-time PCR (qRT-PCR), while being faster and more user-friendly.

Article Abstract

Background: The aim of the study was to evaluate a loop-mediated isothermal amplification (LAMP) assay for the ability to diagnose tuberculosis directly from clinical samples rapidly.

Methods: LAMP assays were performed using previously reported primer sets to amplify three specific Mycobacterium tuberculosis (MTB) gene targets, hspX, gyrB, and IS6110. Quantitated DNA from strain H37Rv were detected for assessment of analytical sensitivity; specificity was evaluated by testing eight species of non-tuberculosis Mycobacterium (NTM) and four unrelated bacterial species. Sputum samples from 68 pulmonary tuberculosis patients and a control group consisting of 45 lung cancer patients and 20 healthy controls were analyzed using LAMP assays, and then compared with smear, culture and quantitative real-time PCR (qRT-PCR) methods.

Results: All three LAMP assays showed 100% specificity for MTB when tested against NTM and other bacterial species. The gyrB-LAMP assay was able to detect 60 cfu/ml of H37Rv suspension within 1 h, similar to qRT-PCR, but 10 times more sensitive than the hspX-LAMP and IS6110-LAMP assays. In clinical samples, when qRT-PCR was used as the reference method, the sensitivity of the three LAMP assays targeting hspX, gyrB, and IS6110 genes was 94.6, 98.2 and 92.9%, respectively.

Conclusions: LAMP is more sensitive than smear microscopy and close to qRT-PCR in sensitivity for the detection of MTB. LAMP has comparable specificity to qRT-PCR but was more rapid and convenient.

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Source
http://dx.doi.org/10.1016/j.mimet.2022.106547DOI Listing

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