A Rapid LAMP-Based Method for Screening Poultry Samples for Campylobacter Without Enrichment

Abstract

Campylobacter is the most prominent bacterium associated with foodborne disease and the majority of human infection cases are attributed to chicken. Rapid methods capable of determining the Campylobacter status of poultry products in a short time are needed in today’s fast-paced food supply chain. In this study, we developed and evaluated an easy to perform, rapid and robust method for direct detection of Campylobacter in poultry carcasses based on loop-mediated isothermal DNA AMPlification (LAMP). The method does not require bacterial culture or DNA purification and generates results in just an hour. A total of 171 swabs from chicken and turkey slaughter houses were analyzed in parallel by both LAMP and conventional culture-based enumeration methods to evaluate the performance of the rapid method. Campylobacter was detected by LAMP in 100% of swabs with an enumeration result of ≥800 cfu/swab, and 98.6% (69 out of 70) of samples reported as negative by enumeration (≤10 cfu/swab) were also negative by LAMP. The method is also suitable for analysis of boot swabs from poultry houses, and therefore it represents a convenient screening tool that can be implemented on farm, at slaughter houses, processing plants or retail, to help with the control of Campylobacter contamination throughout the food supply chain. The inclusion of an internal amplification control prevents any potential false negative results due to DNA amplification inhibitors that might be present in the sample.

A Sensitive, Specific and Simple Loop Mediated Isothermal Amplification Method for Rapid Detection of Campylobacter spp. in Broiler Production

“…LODs of 1 CFU/reaction (corresponding to 50 CFU/ml) were observed for both C. jejuni and C. coli within 30 min of amplification (Figure 4). The optimized LAMP assay, in this study, showed better performance with lower LOD and shorter reaction time than previous reports (1.2–1.4 CFU and 5 CFU per reaction within 60 min) (Yamazaki et al, 2008; Romero and Cook, 2018).…”

“…LAMP was developed for the detection of Campylobacter spp. in poultry samples such as meat, carcass swabs, and fecal samples (Yamazaki et al, 2008, 2009b; Sabike et al, 2016; Romero and Cook, 2018; Sabike and Yamazaki, 2019). To study the epidemiology of Campylobacter spp.…”

One-Day Molecular Detection of Salmonella and Campylobacter in Chicken Meat: A Pilot Study

“…Performing 45 LAMP significantly increased the number of detected positive samples (from 74% to 82.5%, Table S2), with 10 1 CFU/g samples being detected after 4 h of enrichment (Figure 6a). In a study on artificially contaminated swabs, the limit of detection of 60 LAMP was reported between 10 3 and 10 4 CFU/swab [24]. In fact, with our protocol, the increase to 60 of the colorimetric LAMP resulted in the production of false positives, with a decrease in specificity (data not shown); however, the 30 or 45 protocol detected down to 10 3 CFU/g without enrichment.…”

“…Colorimetric LAMP assays were carried out using primers previously described by Zhuang et al [22] for Salmonella and by Romero et al [23,24] for Campylobacter spp. (Table 2).…”

“…Clinical manifestations of campylobacteriosis can be severe, with possible sequelae like Guillain-Barrè syndrome, reactive arthritis, and irritable bowel syndrome [5]. The main reservoir for foodborne transmission of C. coli and C. jejuni are poultry species, and handling and eating raw or undercooked chicken meat is the main risk factor for human infection, accounting for up to 24.2% of all of the Campylobacter spp. infections in the EU [6].…”