Rapid Detection of Viable Escherichia coli O157 by Coupling Propidium Monoazide with Loop-Mediated Isothermal Amplification

2013 ◽  
Vol 23 (12) ◽  
pp. 1708-1716 ◽  
Author(s):  
Xihong Zhao ◽  
Jun Wang ◽  
Fereidoun Forghani ◽  
Joong-Hyun Park ◽  
Myoung-Su Park ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Rapid Detection ◽  
Isothermal Amplification ◽  
Escherichia Coli O157 ◽  
Propidium Monoazide ◽  
Loop Mediated Isothermal Amplification

scholarly journals Visual Detection of Enterohemorrhagic Escherichia coli O157:H7 Using Loop-Mediated Isothermal Amplification

10.19082/2576 ◽  
2016 ◽  
Vol 8 (6) ◽  
pp. 2576-2585 ◽  
Author(s):  
Reza Ranjbar ◽  
Maryam Erfanmanesh ◽  
Davoud Afshar ◽  
Mohsen Mohammadi ◽  
Omar Ghaderi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Visual Detection ◽  
Enterohemorrhagic Escherichia Coli ◽  
Isothermal Amplification ◽  
Escherichia Coli O157 ◽  
Loop Mediated Isothermal Amplification

scholarly journals Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Common Strains of Escherichia coli

2008 ◽  
Vol 46 (8) ◽  
pp. 2800-2804 ◽  
Author(s):  
J. Hill ◽  
S. Beriwal ◽  
I. Chandra ◽  
V. K. Paul ◽  
A. Kapil ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Rapid Detection ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification Assay

scholarly journals Rapid Detection of Viable Salmonellae in Produce by Coupling Propidium Monoazide with Loop-Mediated Isothermal Amplification

2011 ◽  
Vol 77 (12) ◽  
pp. 4008-4016 ◽  
Author(s):  
Siyi Chen ◽  
Fei Wang ◽  
John C. Beaulieu ◽  
Rebecca E. Stein ◽  
Beilei Ge
Keyword(s):  
Pure Culture ◽  
Rapid Detection ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Detection Methods ◽  
Sample Treatment ◽  
Propidium Monoazide ◽  
Simple Method ◽  
Content Type ◽  
Loop Mediated Isothermal Amplification

ABSTRACTRecent outbreaks linked toSalmonella-contaminated produce heightened the need to develop simple, rapid, and accurate detection methods, particularly those capable of determining cell viability. In this study, we examined a novel strategy for the rapid detection and quantification of viable salmonellae in produce by coupling a simple propidium monoazide sample treatment with loop-mediated isothermal amplification (PMA-LAMP). We first designed and optimized a LAMP assay targetingSalmonella. Second, the performance of PMA-LAMP for detecting and quantifying viable salmonellae was determined. Finally, the assay was evaluated in experimentally contaminated produce items (cantaloupe, spinach, and tomato). Under the optimized condition, PMA-LAMP consistently gave negative results for heat-killedSalmonellacells with concentrations up to 108CFU/ml (or CFU/g in produce). The detection limits of PMA-LAMP were 3.4 to 34 viableSalmonellacells in pure culture and 6.1 × 103to 6.1 × 104CFU/g in spiked produce samples. In comparison, PMA-PCR was up to 100-fold less sensitive. The correlation between LAMP time threshold (TT) values and viableSalmonellacell numbers was high (R2= 0.949 to 0.993), with a quantification range (102to 105CFU/reaction in pure culture and 104to 107CFU/g in produce) comparable to that of PMA in combination with quantitative real-time PCR (PMA-qPCR). The complete PMA-LAMP assay took about 3 h to complete when testing produce samples. In conclusion, this rapid, accurate, and simple method to detect and quantify viableSalmonellacells in produce may present a useful tool for the produce industry to better control potential microbial hazards in produce.

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