Development of a loop-mediated isothermal amplification (LAMP) method for rapid detection of salmonella spp in feedstuff

Name: Sithisak Masphol

LCSH: Kasetsart University -- Theses. M.S. (Agricultural Biotechnology) 2021

Classification :.LCCS: QR201.S25

LCSH: Kasetsart University. -- Interdisciplinary Graduate Program -- Theses

LCSH: Salmonella

LCSH: Foodborne diseases

LCSH: Pathogenic bacteria

Abstract: Salmonella spp . is a food-borne pathogenic bacterium that causes food poisoning in humans and animals. The Salmonella causative source has traditionally been attributed to feed ingredients, feed and animal products. Conventional detection methods of Salmonella spp. are time-consuming since they taking time for 5 days. Herein, a Loop mediated isothermal amplification integrated with LAMP-AGE and LAMP-SYBR green I was developed to increase the yield of the Salmonella invasion gene (InvA gene) with SalinvA primer set and identification of Salmonella spp. within approximately 60 minutes with the optimized temperature at 62°C for SalinvA01 and 61°C for SalinvA02-05 primer. Subject to these optimized conditions, the detection limits of the SalinvA01 primer for Salmonella spp. pure culture was 1.33×103 CFU/mL or 26.6 CFU/tube. The specificity showed no cross-reactivity with 15 isolates of non-Salmonella spp. Salmonella contaminated feed and feedstuffs were detected using the LAMP technique after 4 hours of pre-enrichment. Among the 24 samples of feed and feed ingredients that were used to evaluate artificial contamination and enrichment of Salmonella spp. LAMP assay demonstrated 100% accuracy compared to PCR and conventional methods. In conclusion, the SalinvA01 primer confirmed to be highly specific, low detection limits, accurate, fast and convenient. The results can be received in one hour by using isothermal conditions, no need to use thermocycler and the test is inexpensive. Hence, the developed assay can be utilized for on-site rapid screening of Salmonella spp. contamination in feed and feed ingredients.

Kasetsart University. Office of the University Library

Address: Bangkok

Email: tdckulib@ku.ac.th

Name: Wirawan Nuchchanart

Role: Thesis Advisor

Name: Nuanchan Paraksa

Role: Thesis CO-Advisor

Created: 2021

Modified: 2025-07-10

Issued: 2025-07-10

วิทยานิพนธ์/Thesis

application/pdf

URL: https://www.lib.ku.ac.th/KUthesis/2564/sithisak-mas-all.pdf

CallNumber: QR201.S25 .S58

eng

DegreeName: Master of Science

Level: Master's Degree

Descipline: Agricultural Biotechnology

Grantor: Kasetsart University

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