A rapid novel visualized loop-mediated isothermal amplification method for Salmonella detection targeting at fimW gene

Wen, Jing; Gou, Hongchao; Zhan, Zeqiang; Gao, Yuan; Chen, Zhengquan; Bai, Jie; Wang, Shaojun; Chen, Kaifeng; Lin, Quankui; Liao, Ming; Zhang, Jianmin

doi:10.1016/j.psj.2020.03.045

Cited by 22 publications

(9 citation statements)

References 18 publications

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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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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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“…Another bacterial rapid test, the colloidal gold method, has simple operation and short reaction time, but it is prone to false positives or false negatives and has low sensitivity and specificity. To compensate for these shortcoming, research into the rapid detection of pathogenic bacteria using molecular biology techniques (such as PCR, gene chip, strand replacement technologies, autonomous replicating sequences, and others) has made great progress (12)(13)(14)(15)(16)(17)(18)(19); nonetheless, modern detection still requires expensive nucleic acid amplification equipment, highly trained personnel, complex procedures, and a detection time of 3-5 hours. Given these restrictions, it can only be performed in large medical institutions, and its application as a rapid diagnostic method is also limited.…”

Section: Discussionmentioning

confidence: 99%

Rapid detection of Salmonella based on loop-mediated isothermal amplification

Wang

Gao

et al. 2021

Ann Palliat Med

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Background: Salmonella enterica is a zoonotic pathogen of substantial concern to human and animal health and is a leading cause of morbidity and mortality in people worldwide. Loop-mediated isothermal amplification (LAMP) technology is a new type of nucleic acid amplification technology, which has the characteristics of high specificity, high sensitivity, simple operation, convenience, and low cost. This study aims to establish a rapid detection method for Salmonella based on LAMP technology. Methods: Primers were designed for Salmonella's specific conservative invA gene. Through primer screening and optimization of reaction conditions, and a LAMP method for detecting Salmonella with realtime fluorescence and visual observation results was established. The sensitivity and specificity of the method were assessed, and the accuracy was evaluated through the testing of Salmonella-contaminated and noncontaminated clinical samples. Results:The optimal reaction temperature of LAMP was 60-65 ℃, and the optimal reaction time was 25-30 minutes. The detection limits of real-time fluorescence and visual observation were both 1.4 pg/μL. There was no cross-reactivity observed with 22 non-Salmonella species, and the specificity was 100%.Additionally, 30 samples contaminated with Salmonella, 30 samples not contaminated with Salmonella, and 8 clinical samples identified as positive by bacterial culture and microbial mass spectrometry were tested. The positive coincidence rate of the detection system was 97.4% by real-time fluorescence and 89.5% by visual observation, the negative coincidence rate was 100%, and the total coincidence rate was 98.5% and 94.1%, respectively.Conclusions: In the scene of infection, primary hospital, disaster area treatment and other scenarios, the conditions of environment, equipment and personnel was limited, therefore, the established real-time fluorescence and visual lamp method can provide a powerful means for the rapid detection of Salmonella.

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“…Currently, the detection of pathogenic bacteria in clinical practice mainly relies on conventional culture methods, such as the examination of body fluids, blood, and secretions (5,6); the cultivation and identification of pathogenic bacteria; and drug sensitivity tests. In recent years, research into the rapid detection of pathogens using molecular biology methods has made great progress (7)(8)(9)(10). However, this detection requires expensive nucleic acid amplification equipment, with high requirements on laboratories and personnel, complicated procedures, and 3-5 hours of detection time.…”

Section: Discussionmentioning

confidence: 99%

Rapid detection of multiple pathogens by the combined loop-mediated isothermal amplification technology and microfluidic chip technology

Ou¹,

Wang²,

Wang³

et al. 2021

Ann Palliat Med

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A rapid novel visualized loop-mediated isothermal amplification method for Salmonella detection targeting at fimW gene

Cited by 22 publications

References 18 publications

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

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

Rapid detection of Salmonella based on loop-mediated isothermal amplification

Rapid detection of multiple pathogens by the combined loop-mediated isothermal amplification technology and microfluidic chip technology

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