scholarly journals Development of Simple Multiplex Real-Time PCR Assays for Foodborne Pathogens Detection and Identification On Lightcycler

2017 ◽  
Vol 40 (1) ◽  
pp. 53-58 ◽  
Author(s):  
Avo Karus ◽  
Fabrizio Ceciliani ◽  
Armand Sanches Bonastre ◽  
Virge Karus
Keyword(s):  
Real Time ◽  
Foodborne Pathogens ◽  
Real Time Pcr ◽  
Simultaneous Detection ◽  
Melting Curve ◽  
Food Contamination ◽  
Shigella Boydii ◽  
Food Borne Pathogens ◽  
Detection And Identification ◽  
Food Borne

Abstract Most acute intestinal diseases are caused by food-borne pathogens. A fast and simple real-time PCR-based procedure for simultaneous detection of food contamination by any of the five food-borne pathogens: Campylobacter jejuni, Mycobacterium bovis, Enterobacter sakazaki, Shigella boydii, Clostridium perfrigens using multiplex EvaGreen real-time PCR for LightCycler was developed and evaluated. Real-time qPCR showed excellent sensitivity. Tm calling and Melting Curve Genotyping (MCG) were used for analysis of PCR product melting curves. The Melting Curve Genotyping option showed good performance for discrimination of positive samples containing DNA of single pathogen or pathogen mixtures from negative samples.

Developing a multiplex real-time PCR with a new pre-enrichment to simultaneously detect four foodborne bacteria in milk

2019 ◽  
Vol 14 (10) ◽  
pp. 885-898 ◽  
Author(s):  
Moezi Parichehr ◽  
Kargar Mohammad ◽  
Doosti Abbas ◽  
Khoshneviszadeh Mehdi
Keyword(s):  
Real Time ◽  
Foodborne Pathogens ◽  
Real Time Pcr ◽  
Simultaneous Detection ◽  
Pcr Assay ◽  
Reliable Technique ◽  
E Coli ◽  
Foodborne Bacteria ◽  
Multiple Pathogens ◽  
Enrichment Step

Aim: The aim of this study is to formulate a new single nonselective pre-enrichment medium (ELSS) that can support the concurrent growth of four major foodborne pathogens containing E. coli O157: H7, L. monocytogenes, S. aureus and S. enterica serovar Entertidis to develop a multiplex TaqMan Real-time PCR (mRT-PCR). Methods: The mRT-PCR with a new pre-enrichment was carried out for simultaneous detection and quantification of these foodborne bacteria. Results: By using mRT-PCR after 16 h pre-enrichment in ELSS, the detection limit of each pathogen was 1 CFU/25 ml contaminated milk, as well as inclusivity and exclusivity reached 100%. Conclusion: The mRT-PCR assay with pre-enrichment step is a fast and reliable technique for detecting single or multiple pathogens in food products.

scholarly journals Establishment and Application of a Dual TaqMan Real-Time PCR Methodfor Proteus mirabilis and Proteus vulgaris

2020 ◽  
Vol 69 (3) ◽  
pp. 293-300
Author(s):  
RUI YANG ◽  
GUOYANG XU ◽  
XIAOYOU WANG ◽  
ZHICHU QING ◽  
LIZHI FU
Keyword(s):  
Real Time ◽  
Foodborne Pathogens ◽  
Proteus Mirabilis ◽  
Real Time Pcr ◽  
Proteus Vulgaris ◽  
Opportunistic Bacteria ◽  
Food Borne ◽  
Colony Forming Units ◽  
Pcr Method ◽  
Pure Bacterial Cultures

Proteus species are common opportunistic bacteria and foodborne pathogens. The proper detection of Proteus can effectively reduce the occurrence of food-borne public health events. Proteus mirabilis and Proteus vulgaris are the two most important pathogens in the Proteus genus. In this study, a dual TaqMan Real-Time PCR method was established to simultaneously detect and distinguish P. mirabilis and P. vulgaris in samples. The method exhibited good specificity, stability, and sensitivity. Specifically, the minimum detection concentrations of P. mirabilis and P. vulgaris in pure bacterial cultures were 6.08 × 102 colony forming units (CFU)/ml and 4.46 × 102 CFU/ml, respectively. Additionally, the minimum detectable number of P. mirabilis and P. vulgaris in meat and milk was 103 CFU/g. In addition, the method can be used to distinguish between strains of P. mirabilis and P. vulgaris within two hours. Overall, it is a sensitive, easy-to-use, and practical test for the identification and classification of Proteus in food.

scholarly journals Development of a melting-curve based multiplex real-time PCR assay for simultaneous detection of Streptococcus agalactiae and genes encoding resistance to macrolides and lincosamides

2018 ◽  
Vol 18 (1) ◽  
Author(s):  
Eliane Saori Otaguiri ◽  
Ana Elisa Belotto Morguette ◽  
Alexandre Tadachi Morey ◽  
Eliandro Reis Tavares ◽  
Gilselena Kerbauy ◽  
...  
Keyword(s):  
Real Time ◽  
Streptococcus Agalactiae ◽  
Real Time Pcr ◽  
Simultaneous Detection ◽  
Melting Curve ◽  
Pcr Assay ◽  
Genes Encoding
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