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

Abstract

Salmonella and Campylobacter ssp. are bacterial pathogens responsible for most foodborne infections in EU countries. Poultry serves as a reservoir for these pathogens, and its important role in the meat industry makes it essential to develop a rapid detection assay able to provide results in one day. Indeed, the rapid identification of foodborne pathogens is an important instrument for the monitoring and prevention of epidemic outbreaks. To date, Salmonella and Campylobacter screening is mainly conducted through molecular methods (PCR or real-time PCR) performed after 18–24 h long enrichments. In this study, we evaluated short enrichments (0, 2, 4, and 6 h) combined with a colorimetric loop-mediated isothermal AMPlification (LAMP) or real-time PCR to detect Salmonella and Campylobacter in poultry meat contaminated at different concentration levels (101, 103, and 105 CFU/g). Our results show that real-time PCR allows the detection of Salmonella and Campylobacter, even after shorter enrichment times than prescribed by ISO references; particularly, it detected Salmonella down to 101 CFU/g since T0 and Campylobacter from 103 CFU/g since T0. Detection with LAMP was comparable to real-time PCR without the requirement of a thermal cycler and with shorter execution times. These characteristics make colorimetric LAMP a valid alternative when one-day results are needed, improving the timely identification of positive meat batches, even in the absence of specialized instrumentation.

Rapid Oxford Nanopore Technologies MinION Sequencing Workflow for Campylobacter jejuni Identification in Broilers on Site—A Proof-of-Concept Study

“…Undoubtedly, it seems essential to assess the status of the flocks before they reach the slaughterhouse [ 30 , 60 , 61 , 62 , 63 , 64 ]. This is why molecular methods for same-day diagnosis of pathogenic bacteria are increasingly being introduced into daily laboratory routines [ 53 , 65 , 66 ]. Overall, 100% of samples were positive for both methods, regardless of animal age, requiring a total of five days from enumeration to confirmation for ISO 1027-2:2017, while the sequencing method required only five hours between extraction and sequencing.…”

Rapid detection methods for foodborne pathogens based on nucleic acid amplification: Recent advances, remaining challenges, and possible opportunities

“…1 B presents the schematic representation of the LAMP amplification. The LAMP primer set includes internal primers (FIP and BIP) as well as external primers (F3 and B3) ( Du et al, 2022 , Jia et al, 2023 , Jiang et al, 2020 , Li et al, 2021 , Priya et al, 2020 , Wang et al, 2020a , Xie et al, 2022a , Xiong et al, 2020 , Zendrini et al, 2021 , Zhang et al, 2021 , Zhang et al, 2022 ). FIP comprises the F1c and F2 regions, while BIP comprises the B1c and B2 regions.…”

“…In recent years, a great number of studies have been devoted to exploring the capability of LAMP technology in detecting various pathogens in various types of foods, including milk, seafood, and meats ( Du et al, 2022 , Jia et al, 2023 , Jiang et al, 2020 , Li et al, 2021 , Priya et al, 2020 , Wang et al, 2020a , Xie et al, 2022a , Xiong et al, 2020 , Zendrini et al, 2021 , Zhang et al, 2021 , Zhang et al, 2022 ). Most of these studies used fluorescence to indicate the presence of pathogens in the amplified product and some of them applied necessary pre-treatment on their samples before DNA amplification ( Table S3 ).…”

“…Interestingly, several of these studies reported that LAMP technology enabled direct recognition of signals with the naked eye, which is probably suitable for on-site detection. For example, the presence of Salmonella or Campylobacter in chicken meat could be detected by observing the turbidity anomaly or colorimetric changes during the amplification process ( Wang et al, 2020a , Zendrini et al, 2021 ). Similarly, Li et al (2021) also reported that this method enabled the detection of Salmonella in pork by simply observing the colorimetric changes with the naked eye.…”

Evaluation of a novel lysis-based sample processing method to optimize Vibrio vulnificus detecting by Loop-Mediated Isothermal Amplification assay

“…Based on the strand displacement reaction, LAMP technology could exploit the ampli cation and detection of speci c DNA sequences under isothermal conditions using Bst DNA polymerase as a strategic tool. Compared with other common detection technologies, LAMP technology has weakened the instruments' requirements and shortened the detection time during a much simpler procedure while optimizing the detection results, such as speci city and sensitivity (Yamazaki et al 2008;Zendrini et al 2021). Due to the abovementioned advantages, LAMP has been gradually developed as the rapid pathogen diagnosis method successfully used to detect Vibrio, Staphylococcus aureus, Novel coronavirus, Hepatitis B virus, and other microorganisms (Chaouch 2021;Choopara et al 2021;Vanhomwegen et al 2021;Zheng et al 2021).…”