Development and evaluation of real‐time loop mediated isothermal amplification assay for rapid and sensitive detection of Salmonella spp. in chicken meat products

Abstract

Salmonella spp., one of the most prevalent bacterial causes of foodborne disease, is a major public health concern due to its common occurrence in chicken meat products. The detection of Salmonella spp. using conventional culturing methods requires an excessive amount of time and effort. Thus, in response to the need for a rapid, sensitive, and convenient method for Salmonella detection, a quantitative assay using real‐time Loop‐mediated isothermal amplification (real‐time LAMP) was developed using gene62181533 as the target sequence amplified. The real‐time LAMP assay does not show cross‐reactivity with several other common bacterial pathogens and the detection limit for genomic DNA in pure culture was found to be 1.2 CFU/ml. Testing with spiked samples simultaneously detected 7 CFU/ml of Salmonella pathogen in the artificially inoculated samples after enrichment for 6 hr. In comparison with the standard culture‐based methods in the analysis of 120 raw chicken meat samples, the results of the sensitivity, accuracy, and specificity tests of the real‐time LAMP assay were 94.02, 90.83, and 86.79%, respectively. These results indicate that this method is not only sensitive and specific but can also be used for rapid detection and differentiation of foodborne disease‐causing bacteria. Practical applications Salmonella is an important bacterial genus which causes most of the common foodborne illnesses worldwide. The disease mainly caused by Salmonella spp. through consumption of contaminated eggs and poultry products is salmonellosis which can manifest as bacterial diarrhea through to septicemia. The result indicates the potential usefulness of real‐time LAMP assay based on the gene62181533 for a rapid, specific, sensitive, and quantitative assay for Salmonella detection in food products. This assay offers a low cost quantitative method in the clinical diagnosis of Salmonella in resource‐limited health‐care facilities and clinical laboratories in developing countries and in field tests.

Detection of Bacterial Pathogens and Antibiotic Residues in Chicken Meat: A Review

“…Arunrut et al [ 82 ] developed a real-time loop-mediated isothermal amplification (LAMP) procedure for the identification of Salmonella spp. with the help gene62181533 as a primer sequence.…”

Rapid detection of Escherichia coli O157:H7 in milk, bread, and jelly by lac dye coloration‐based bidirectional lateral flow immunoassay strip

“…The detection methods for foodborne pathogens were based on molecular biology and immunology. The molecular biology‐based methods incorporate polymerase chain reaction (PCR) (Hull‐Jackson, Mota‐Meira, & Adesiyun, 2019), real‐time PCR (Alia, Andrade, Cordoba, Martin, & Rodriguez, 2020; Zhong, Tian, Wang, & Wang, 2016), multiplex PCR (Chen, Tang, Liu, Cai, & Bai, 2012; Wei et al, 2019; Xu et al, 2019), loop‐mediated isothermal amplification (LAMP) (Arunrut, Kiatpathomchai, & Ananchaipattana, 2018; Kampeera et al, 2019; Wan, Lu, Bie, Lv, & Zhao, 2020), recombinase polymerase amplification (RPA) (Ren et al, 2020), and rolling circle amplification (RCA) (Jiang et al, 2019). Although these methods can achieve high sensitivity, the requirement of high‐quality professional technicians and instruments limit the use in special places.…”