Development of real-time Loop-mediated isothermal amplification assay for rapid detection of Salmonella enterica serovars

Authors

Abstract

Typhoid fever remains a major global health burden, particularly in regions with inadequate sanitation and safe water supplies. The disease, caused by Salmonella enterica serovars transmitted via the fecal-oral route, can lead to severe systemic infections with life-threatening complications and contribute to chronic carrier states that perpetuate its spread. Traditional diagnostic methods such as blood culture and biochemical assays, though considered gold standards, are labor-intensive and time-consuming, thereby limiting their practical utility in resource-constrained settings. In response, we developed a real-time loop-mediated isothermal amplification (LAMP) assay for the rapid detection of S. enterica using conserved genomic regions as targets. Unlike conventional PCR, which requires expensive thermal cyclers, our LAMP protocol utilizes a water bath to maintain a constant reaction temperature of 63°C for 90 minutes, significantly reducing equipment costs. Amplified products were rapidly detected by the addition of SYBR Green dye, which produced a distinct green fluorescence under blue light, enabling easy visual interpretation. Comparative analysis revealed that the LAMP assay demonstrated superior sensitivity—detecting Salmonella in 90% of isolates—compared to 85% for PCR, with both methods showing high specificity and no cross-reactivity with non-Salmonella species. These results indicate that the LAMP assay offers a rapid, cost-effective, and sensitive alternative for typhoid diagnosis, holding promise for improved disease surveillance and management in low-resource settings.

Author Biographies

Yusuf Musa Ibrahim, Federal University Dutse

Department of Microbiology and Biotechnology

Abdulaziz Dantata, Federal University Dutse

Department of Microbiology and Biotechnology

Tsoho Rabiu Nura, Federal University Dutse

Department of Microbiology and Biotechnology

Ahmad Mohammed Gumel, Federal University Dutse

Department of Microbiology and Biotechnology

Published

2025-04-26

How to Cite

Muhtar, U. N., Ibrahim, Y. M., Dantata, A., Nura, T. R., & Gumel, A. M. (2025). Development of real-time Loop-mediated isothermal amplification assay for rapid detection of Salmonella enterica serovars. UMYU Conference of Microbiology and Related Sciences, 1(1). Retrieved from https://ujmr.umyu.edu.ng/index.php/mcbconference/article/view/974