scholarly journals Rapid and simultaneous detection of Campylobacter spp. and Salmonella spp. in chicken samples by duplex loop-mediated isothermal amplification coupled with a lateral flow biosensor assay

PLoS ONE ◽  
2021 ◽  
Vol 16 (7) ◽  
pp. e0254029
Author(s):  
Thanawat Sridapan ◽  
Wanida Tangkawsakul ◽  
Tavan Janvilisri ◽  
Taradon Luangtongkum ◽  
Wansika Kiatpathomchai ◽  
...  
Keyword(s):  
Simultaneous Detection ◽  
Chicken Meat ◽  
Lateral Flow ◽  
Isothermal Amplification ◽  
Salmonella Spp ◽  
Promising Alternative ◽  
Loop Mediated Isothermal Amplification ◽  
Alternative Approach ◽  
Meat Samples ◽  
Sensitivity Specificity

Development of a simple, rapid and specific assay for the simultaneous detection of Campylobacter spp. and Salmonella spp. based on duplex loop-mediated isothermal amplification (d-LAMP), combined with lateral-flow biosensor (LFB) is reported herein. LAMP amplicons of both pathogens were simultaneously amplified and specifically differentiated by LFB. The specificity of the d-LAMP-LFB was evaluated using a set of 68 target and 12 non-target strains, showing 100% inclusivity and exclusivity. The assay can simultaneously detect Campylobacter and Salmonella strains as low as 1 ng and 100 pg genomic DNA per reaction, respectively. The lowest inoculated detection limits for Campylobacter and Salmonella species in artificially contaminated chicken meat samples were 103 CFU and 1 CFU per 25 grams, respectively, after enrichment for 24 h. Furthermore, compared to culture-based methods using field chicken meat samples, the sensitivity, specificity and accuracy of d-LAMP- LFB were 95.6% (95% CI, 78.0%-99.8%), 71.4% (95% CI, 29.0%-96.3%) and 90.0% (95% CI, 73.4%-97.8%), respectively. The developed d-LAMP-LFB assay herein shows great potentials for the simultaneous detection of the Campylobacter and Salmonella spp. and poses a promising alternative approach for detection of both pathogens with applications in food products.

The Detection of Tuberculosis by Loop-Mediated Isothermal Amplification (LAMP) Combined with a Lateral Flow Dipstick

2015 ◽  
pp. 269-300
Author(s):  
Thongchai Kaewphinit ◽  
Somchai Santiwatanakul ◽  
Kosum Chansiri
Keyword(s):  
Fluorescein Isothiocyanate ◽  
Lateral Flow ◽  
Ease Of Use ◽  
Isothermal Amplification ◽  
Toxic Substances ◽  
Nested Polymerase Chain Reaction ◽  
Loop Mediated Isothermal Amplification ◽  
Lateral Flow Dipstick ◽  
Electrophoresis Method ◽  
Sensitivity Specificity

Tuberculosis (TB) is an airborne infectious disease caused by the bacterium Mycobacterium Tuberculosis (MTB) and is a persistent problem in developing countries. Present methods for its detection include normal or nested Polymerase Chain Reaction (PCR) followed by electrophoresis, real-time PCR, Ziehl-Neelsen staining, and culture assay. These techniques entail various disadvantages such as high cost, long assay time and use of toxic substances. Novel loop-mediated isothermal amplification (LAMP) permits DNA to be amplified rapidly under constant temperature. The combination of LAMP and chromatographic Lateral Flow Dipstick (LAMP-LFD) by using biotinylated LAMP amplicon hybridized with Fluorescein Isothiocyanate (FITC)-labeled probes are allowed to detect MTB without electrophoresis and interpreted within 3-5 min. LAMP-LFD is as highly sensitive as PCR-electrophoresis method. Based on its sensitivity, specificity, rapidity, cost effectiveness, ease of use, and convenience, LAMP-LFD could be suitable for use in early MTB detection.

scholarly journals Comparison of Loop-Mediated Isothermal Amplification Assay and Conventional Culture Methods for Detection of Campylobacter jejuni and Campylobacter coli in Naturally Contaminated Chicken Meat Samples

2009 ◽  
Vol 75 (6) ◽  
pp. 1597-1603 ◽  
Author(s):  
Wataru Yamazaki ◽  
Masumi Taguchi ◽  
Takao Kawai ◽  
Kentaro Kawatsu ◽  
Junko Sakata ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Lamp Assay ◽  
Chicken Meat ◽  
Isothermal Amplification ◽  
Campylobacter Coli ◽  
Culture Methods ◽  
Culture Test ◽  
Loop Mediated Isothermal Amplification ◽  
Conventional Culture ◽  
Meat Samples

ABSTRACT We investigated the efficacy of a loop-mediated isothermal amplification (LAMP) assay for detection of chicken meat samples naturally contaminated with Campylobacter jejuni and Campylobacter coli. A total of 144 Preston enrichment broth cultures from chicken meat samples were assessed by using the LAMP assay and conventional culture methods, which consist of a combination of Preston enrichment culturing and plating onto Butzler and modified charcoal cefoperazone deoxycholate agars. Compared with C. jejuni-C. coli isolation using the conventional culture test, the LAMP results showed 98.5% (67/68) and 97.4% (74/76) sensitivity and specificity, respectively, and the positive and negative predictive values were 97.1% (67/69) and 98.7% (74/75), respectively. The conventional culture test required more than 3 to 4 days to isolate and identify C. jejuni and C. coli in the Preston enrichment cultures. In contrast, the LAMP assay was markedly faster, requiring less than 90 min from the beginning of DNA extraction to final detection and differentiation of C. jejuni and C. coli. In total, the LAMP assay required 23.5 to 25.5 h from the beginning of the enrichment culture to final determination. These results suggest that our LAMP assay is a powerful tool for rapid, sensitive, and practical detection of C. jejuni and C. coli which may facilitate surveillance and control of C. jejuni-C. coli contamination in chicken, as well as investigations of food poisoning incidents caused by these organisms. This is the first report of a highly sensitive and specific LAMP assay to detect and differentiate C. jejuni and C. coli in chicken meat samples.

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