The performance of an in-house loop-mediated isothermal amplification for the rapid detection of Mycobacterium tuberculosis in sputum samples in comparison with Xpert MTB/RIF, microscopy and culture

Abstract

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Evaluation of an in-house loop-mediated isothermal amplification for Mycobacterium tuberculosis detection in a remote reference laboratory, Thailand

“…According to the literature, the sensitivity of this inhouse LAMP reported in the present study was similar to the majority of the previous studies in comparison to the conventional PCR, Xpert MTB/RIF and culture [10][11][12] , and similar to that reported in the study carried out in the community hospital (88.8%) 17 . Whereas, the sensitivities of Anyplex plus MTB/NTM PCR using culture as a reference standard were reported at 89.0% 5 , and 87.5% 18 .…”

“…The stored remnant DNA was retrieved for the first time and further analyzed by the in-house LAMP test. The LAMP test was performed using a set of LAMP primers, the reaction mixture, and the condition as described previously 11 , 12 . Briefly, the reaction mixture was prepared using 12 μL of LAMP master mixture (2 μL 10X buffer, 4 μL 2mM dNTP each, 3.2 μL 5M betaine, 1.2 μL 100 mM MgSO 4 , 1.6 μL primer mixture), 1 μL Bst DNA polymerase enzyme (New England Biolabs, Ipswich MA, USA), and 1 μL Calcein fluorescent detection reagent 14 .…”

“…We applied our LAMP assay and compared the results with those of culture as a reference test. The assay showed a high sensitivity (94.7%) and specificity (100%) for MTB detection, in comparison to the Xpert MTB/RIF 11 . In addition, its sensitivity was comparable with the conventional PCR and mycobacterial culture 12 , and its simplicity and cost-effectiveness was attractive for clinical laboratories.…”

Performance of Loop-Mediated Isothermal Amplification (LAMP)-Lateral Flow Detection and Xpert MTB/RIF Ultra for Diagnosis of Pulmonary Tuberculosis

“…They analyzed 26 studies and found that the LAMP assay had a sensitivity and specificity of 89.6% (95% CI 85.6% -92.6%) and 94% (95% CI 91% -96.1%). Likewise, in Thailand, Phetsuksiri et al [8] revealed that simultaneous DNA hybridization and LAMP integrated with LFD had high sensitivity S. Kounmee et al for detecting TB. When compared with the Xpert MTB/RIF assay using clinical samples, the sensitivity values were 82.1% (95% CI 75.4% -88.98%) and 86.9% (95% CI 80.5% -90.8%), respectively, similar to the results of this study.…”

Combining LAMP and Au-Nanoprobe to detect INH-RIF resistance accurately in tuberculosis: an evidence-based review

“…On the other hand, studies using LAMP without Au-Np as a diagnostic tool are abundant and show the applicability of LAMP in different countries of different conditions. In several multicenter studies, LAMP is applicable in China [42,61,63,66,83,86], India [74,77,84,87], Sri Lanka [79], Nepal [85], Thailand [12,15,72,88], Vietnam [70,76], Indonesia [90], Malawi [71], South Africa [76,78], Ethiopia [75], Gambia [80], Zambia [73], Uganda [82,84], Peru [76,84], Brazil [76], Japan [68,89], and Korea [81]. Most studies about LAMP were carried out in developing countries that aim to solve the diagnostic problem of TB in burdened countries.…”