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Bacteriology

Optimization of the boil loop-mediated isothermal amplification (LAMP) method for detection of Mycobacterium tuberculosis in saliva

Patricia Gita Naully Medical Laboratory Technology Study Program (Diploma IV), Faculty of Health Sciences and Technology, Jenderal Achmad Yani University, Cimahi, Indonesia., Indonesia Correspondence patriciagitanaully@gmail.com
,
Nur Andini Khoirunisa Medical Laboratory Technology Study Program (Diploma IV), Faculty of Health Sciences and Technology, Jenderal Achmad Yani University, Cimahi, Indonesia., Indonesia Correspondence khandinin@gmail.com
,
Sitti Romlah Medical Laboratory Technology Study Program (Diploma III), Faculty of Health Sciences and Technology, Jenderal Achmad Yani University, Cimahi, Indonesia., Indonesia Correspondence keemov@gmail.com
Pages 386-394
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Abstract

Indonesia has the second-highest tuberculosis (TB) burden worldwide. Conventional TB diagnostic methods in Indonesia are limited by sensitivity, processing time, and cost. The Boil Loop-Mediated Isothermal Amplification (Boil LAMP) method using saliva offers a potential alternative. This study aimed to determine the optimal conditions of the Boil LAMP method for detecting the gyrB gene of Mycobacterium tuberculosis in saliva. Saliva samples were collected from 15 TB patients in Cimahi, Indonesia. Genomic DNA was extracted using simple and modified boiling methods. Optimization of the LAMP assay was performed using genomic DNA concentrations of 25, 50, and 100 ng/µl and amplification temperatures of 59–62°C. Amplification products were analyzed by agarose gel electrophoresis. The modified boiling method yielded salivary genomic DNA with an average concentration of 139 ng/µl and a purity of 1.8. The 320 bp gyrB gene was successfully amplified, with the strongest amplification observed at a DNA concentration of 100 ng/µl and a temperature of 61°C. In conclusion, the modified boiling method combined with the LAMP assay using 100 ng/µl genomic DNA at 61°C provides optimal conditions for detecting M. tuberculosis in saliva, offering a rapid, non-invasive, and cost-effective diagnostic approach for TB.

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How to Cite This

Naully, P. G., Khoirunisa, N. A., & Romlah, S. (2025). Optimization of the boil loop-mediated isothermal amplification (LAMP) method for detection of Mycobacterium tuberculosis in saliva. Jurnal Teknologi Laboratorium, 14(2), 386–394. https://doi.org/10.29238/teknolabjournal.v14i2.365

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