Molecular Detection of <i>Mycobacterium Leprae</i> Using RLEP Loop-Mediated Isothermal Amplification and a Restriction Enzyme to Ensure Amplification Specificity

Early and accurate diagnosis of leprosy is important but remains a significant challenge. Loop-mediated isothermal amplification (LAMP) is a process for the amplification of nucleic acids at a constant temperature and has been used to develop field-friendly tests for many diseases. In the present study, we describe the development of a colorimetric LAMP assay targeting Mycobacterium leprae-specific 450 bp conserved region of the repeat sequences known as RLEP. In addition, the amplicons of LAMP were subjected to restriction analysis using the enzyme EcoRV for specificity. This method has the potential to become an accurate and efficient alternative to Sanger sequencing, which is currently used to validate RLEP-amplified products.

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Journal
Japanese Journal of Infectious Diseases
Published
2024-12-26
DOI
https://doi.org/10.7883/yoken.jjid.2024.251
Primary Topic
Mycobacterium research and diagnosis
Type
article
Field-Weighted Citation Impact
0.00

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article

Molecular Detection of Mycobacterium Leprae Using RLEP Loop-Mediated Isothermal Amplification and a Restriction Enzyme to Ensure Amplification Specificity

Srishti Tripathi, Mukul Sharma, Purna Dwivedi, Pushpendra Singh et al.
Japanese Journal of Infectious Diseases
Mycobacterium research and diagnosis
article

Molecular Detection of Mycobacterium Leprae Using RLEP Loop-Mediated Isothermal Amplification and a Restriction Enzyme to Ensure Amplification Specificity

Srishti Tripathi, Mukul Sharma, Purna Dwivedi, Pushpendra Singh, Purushottam Patel
article en

Abstract

Early and accurate diagnosis of leprosy is important but remains a significant challenge. Loop-mediated isothermal amplification (LAMP) is a process for the amplification of nucleic acids at a constant temperature and has been used to develop field-friendly tests for many diseases. In the present study, we describe the development of a colorimetric LAMP assay targeting Mycobacterium leprae-specific 450 bp conserved region of the repeat sequences known as RLEP. In addition, the amplicons of LAMP were subjected to restriction analysis using the enzyme EcoRV for specificity. This method has the potential to become an accurate and efficient alternative to Sanger sequencing, which is currently used to validate RLEP-amplified products.

Japanese Journal of Infectious DiseasesVol. 78(5)
Maharaja Sayajirao University of Baroda (IN), Netaji Subhash Chandra Bose Medical College (IN), National Institute of Infectious Diseases (JP), National Institute for Research in Tribal Health (IN), Society for Education Welfare and Action Rural (IN), Academy of Scientific and Innovative Research (IN)
Indian Council of Medical Research
Openalex Percentile: Top 64%
Mycobacterium research and diagnosis
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