Quantitative LAMP and PCR Detection of <i>Salmonella</i> in Chicken Samples Collected from Local Markets around Pathum Thani Province, Thailand

Vichaibun, Virun; Kanchanaphum, Panan

doi:10.1155/2020/8833173

Cited by 20 publications

(11 citation statements)

References 28 publications

(32 reference statements)

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“…Moreover, the 45 assay was positive for the 10 1 CFU/g samples after a 2 h enrichment, and even without enrichment in batch 1 (Figure 4a,b). These results were consistent with a recent study, which detected after 2 h of enrichment all of the spiked samples as 100% positive by conventional 45 LAMP [25], while, without the enrichment step, the reported sensitivity varied from 2.2 CFU/g to 10 8 CFU/mL [8]. However, compared to conventional LAMP, the colorimetric assay enables an easy detection of positive samples without any additional processing or specialist interposition, allowing field-based diagnostics.…”

Section: Discussionsupporting

confidence: 89%

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

Zendrini

Carta

Filipello

et al. 2021

Foods

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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.

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Section: Discussionsupporting

confidence: 89%

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

Zendrini

Carta

Filipello

et al. 2021

Foods

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“…All LAMP reaction were performed followed the methods of Kanchanaphum and Vichaibun [ 20 ] containing 5 mM MgSO 4 , 400 mM betaine (Sigma), 1.2 mM dNTPs, 0.8 μ M F3 and B3 primers, 2 μ M FIP and BIP primers, and 8 U Bst DNA polymerase (New England Biolabs). After the LAMP reaction, the products were analyzed by electrophoresis using 1.5% agarose gel and were analyzed by Gel Doc TM XR+ with Image Lab TM Software (BIO RAD, USA.).…”

Section: Methodsmentioning

confidence: 99%

“…For the experiment, the peanut and dried shrimp samples were transferred to a sterile container, and then, the methods of Kanchanaphum and Vichaibun [ 20 ], Shapira et al [ 23 ], and Siruguri et al [ 24 ] were followed. Then, all samples were dried in a laminar hood under ultraviolet light for 3 min.…”

Section: Methodsmentioning

confidence: 99%

“…Nowadays, LAMP has become the interesting method to replace PCR due to its more rapid, simple, and specificity reaction. In this present time, LAMP has been developed and used in many applications, such as for sex determination in human [ 17 ], detection of pork meat in Halal food [ 18 ], detection of Yersinia enterocolitica in pork meat [ 19 ], and detection of Salmonella infection in chicken samples [ 20 ] This method was performed under isothermal conditions in the temperature range 60°C–65°C for 60 minutes [ 21 ]. There are two sets of primer; inner primer and outer primer sets used in LAMP were specific at six different DNA sequences within the target DNA, and primary DNA amplification was begun by the inner primer set.…”

Section: Introductionmentioning

confidence: 99%

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A Comparison of Four Molecular Methods for Detection of Aflatoxin-Producing Aspergillus in Peanut and Dried Shrimp Samples Collected from Local Markets around Pathum Thani Province, Thailand

Kumsiri

Kanchanaphum

2020

Scientifica

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Aspergillus flavus is an aflatoxin-producing fungus which is poisonous to humans and animals when consumed. Detecting the fungus can help to prevent this danger. The four molecular methods, namely, conventional isothermal amplification (LAMP), PCR, quantitative LAMP (qLAMP), and qPCR, were compared to determine their efficiency for A. flavus detection. Thirty samples of peanut and dried shrimp were collected from 15 markets around Pathum Thani Province in Thailand. The samples were artificially infected with 108 conidia/ml of A. flavus for 1 hr and enriched for one day to represent real contamination. The results show that the sensitivity detection for A. flavus in PCR, LAMP, qPCR, and qLAMP was 50 ng, 5 ng, 5 pg, and 5 pg, respectively. Aspergillus in 30 peanut and dried shrimp from the market was detected by all four methods. The detection rate was about 20%, 60%, 100%, and 100% with PCR, LAMP, qPCR, and qLAMP, respectively. The molecular detection technique, especially LAMP, qPCR, and qLAMP, can detect this pathogenic fungi very rapidly with high sensitivity and reliability in comparison to conventional PCR.

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“…Several Salmonella LAMP assays, mostly targeting the invA gene, have been applied in various food and feed matrices, including chicken, turkey, pork, beef, and produce, as shown in a comprehensive review (50) and a few recent studies (18,47,48). Using spiked samples, one assay was able to directly detect (i.e., no enrichment) 10 2 CFU/mL Salmonella in raw beef, pork, chicken, lettuce, vegetable salad, watermelon, and other foods (39), whereas another assay reached the same level of detection in raw pork and milk with 24-h enrichment (19).…”

Section: Discussionmentioning

confidence: 99%

Rapid Screening for Salmonella in Raw Pet Food by Loop-Mediated Isothermal Amplification

Domesle

Young

2021

Journal of Food Protection

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Raw pet food, comprised of raw meat and vegetables, has increased in popularity in recent years. Multiple surveys and frequent recalls indicate that this commodity has a high risk of contamination with Salmonella and other foodborne pathogens. Improved screening methods are needed to meet the growing demand for testing. This matrix verification study aimed to apply a Salmonella loop-mediated isothermal amplification (LAMP) method, recently completed multi-laboratory validation in dry dog food, in several raw pet food matrices, following the U.S. Food and Drug Administration’s method validation guidelines. Five types of raw pet food, consisting of freeze-dried beef and chicken treats, and frozen beef, pork, and turkey complete foods, were evaluated. For each matrix, two sets of ten 25-g test portions (seven inoculated with ≤ 30 cells of Salmonella Typhimurium and three uninoculated controls) were examined. One set was preenriched in buffered peptone water and the other one in lactose broth, which was followed by LAMP screening using two isothermal master mixes (ISO-001 and ISO-004). All results were confirmed by culture as specified in the Bacteriological Analytical Manual (BAM). The LAMP method accurately detected Salmonella in all inoculated test portions of the five raw pet food samples, regardless of the preenrichment broth used. Positive results could be obtained within 4 min of the LAMP run using the LAMP ISO-004 master mix. All uninoculated controls tested negative by LAMP or BAM. Additionally, one turkey-based complete pet food sample was found to be already contaminated with three Salmonella serovars harboring multiple antimicrobial resistance genes. The Salmonella LAMP method offers a rapid, reliable, and robust tool for routine screening of Salmonella in raw pet food, which will help better ensure product safety and protect public health.

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Quantitative LAMP and PCR Detection of Salmonella in Chicken Samples Collected from Local Markets around Pathum Thani Province, Thailand

Cited by 20 publications

References 28 publications

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

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

A Comparison of Four Molecular Methods for Detection of Aflatoxin-Producing Aspergillus in Peanut and Dried Shrimp Samples Collected from Local Markets around Pathum Thani Province, Thailand

Rapid Screening for Salmonella in Raw Pet Food by Loop-Mediated Isothermal Amplification

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