Detection of DNA from Escherichia coli, Clostridium perfringens, Staphylococcus aureus and Bacillus cereus after simplified enrichment using a novel multiplex real-time PCR system

2016 ◽  
Vol 243 (3) ◽  
pp. 521-530
Author(s):  
René Köppel ◽  
Ingrid Tolido ◽  
Gabriela Marti ◽  
Martin Peier
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Bacillus Cereus ◽  
Real Time ◽  
Clostridium Perfringens ◽  
Real Time Pcr

scholarly journals Application of a Real-Time PCR Method for Salmonella spp., Escherichia coli, Staphylococcus aureus and Clostridium perfringens Detection in Water Samples

2013 ◽  
Vol 62 (4) ◽  
pp. 439-443
Author(s):  
BEATA ROZWADOWSKA ◽  
MARTA ALBERTYŃSKA ◽  
GRZEGORZ HUDZIK ◽  
HUBERT OKŁA ◽  
KRZYSZTOF P. JASIK ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Real Time ◽  
Clostridium Perfringens ◽  
Water Samples ◽  
Real Time Pcr ◽  
Culture Method ◽  
Salmonella Spp ◽  
Detection And Identification ◽  
Pcr Method

The diagnostic assessment of water sanitary state is based mainly on the cultivation of bacteria retained on membrane filters. However classical microbiology methods have a lot of disadvantages. More and more frequently, rapid detection and identification of pathogens present in water is based on molecular biology techniques. The aim of this study was to determine the effectiveness and usefulness of a real-time PCR method, when compared to the recommended bacteria culture method, in diagnostics of pathogens in water samples. The research concerned the detection and identification of main sanitary indicators of water such as: Salmonella spp., Escherichia coli, Staphylococcus aureus and Clostridium perfringens. The analyses were conducted in water samples contaminated with the reference material (the aforementioned bacteria) and real environmental samples, which were examined for the presence of nucleic acid of: Salmonella spp., E. coli, S. aureus and C. perfringens using a real-time PCR method.

Fate of Pathogens Inoculated onto Vacuum-Packaged Sliced Hams to Simulate Contamination During Packaging1

1979 ◽  
Vol 42 (6) ◽  
pp. 464-469 ◽  
Author(s):  
M. E. STILES ◽  
L.-K. NG
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Salmonella Typhimurium ◽  
Bacillus Cereus ◽  
Clostridium Perfringens ◽  
Subsequent Treatment ◽  
Vacuum Packaging ◽  
E Coli ◽  
Survival And Growth ◽  
And Staphylococcus Aureus

Ham and chopped ham from two manufacturers were contaminated with five enteropathogens: Bacillus cereus, Clostridium perfringens, Escherichia coli, Salmonella typhimurium and Staphylococcus aureus, at time of slicing and vacuum-packaging, to simulate contamination by manufacturer. Subsequent treatment of the samples, representing sound and undesirable retail handling and consumer use conditions, indicated marked differences in the fate of the pathogens between these products and within product type between the two manufacturers. Greatest differences were observed between the chopped ham products. All pathogens, except C. perfringens, grew actively in fresh ham and chopped ham with abusive holding at 30 and 21 C. After storage at 4 or 10 C for 30 days, B. cereus and C. perfringens were no longer detected, even after subsequent holding at 30 or 21 C for 24 h. E. coli survival and growth was variable, S. typhimurium survived well and grew under some conditions and S. aureus was generally inhibited at high levels of competition.

A Rapid Multiplex Real-Time PCR High-Resolution Melt Curve Assay for the Simultaneous Detection of Bacillus cereus, Listeria monocytogenes, and Staphylococcus aureus in Food

2016 ◽  
Vol 79 (5) ◽  
pp. 810-815 ◽  
Author(s):  
FEREIDOUN FORGHANI ◽  
SHUAI WEI ◽  
DEOG-HWAN OH
Keyword(s):  
Staphylococcus Aureus ◽  
Listeria Monocytogenes ◽  
High Resolution ◽  
Bacillus Cereus ◽  
Real Time ◽  
Real Time Pcr ◽  
Simultaneous Detection ◽  
Cost Savings ◽  
High Resolution Melt ◽  
And Staphylococcus Aureus

ABSTRACTThree important foodborne pathogens, Bacillus cereus, Listeria monocytogenes, and Staphylococcus aureus, are of great concern for food safety. They may also coexist in food matrices and, in the case of B. cereus and S. aureus, the resulting illnesses can resemble each other owing to similar symptoms. Therefore, their simultaneous detection may have advantages in terms of cost savings and rapidity. Given this context, a rapid multiplex real-time PCR high-resolution melt curve assay for the simultaneous detection of these three pathogens in food was developed. The assay successfully detected B. cereus (gyrB), L. monocytogenes (hly), and S. aureus (nuc) in a single reaction, and the average melting temperatures were 76.23, 80.19, and 74.01°C, respectively. The application of SYTO9 dye and a slow melt curve analysis ramp rate (0.1°C/s) enabled the production of sharp, high-resolution melt curve peaks that were easily distinguishable from each other. The detection limit in food (milk, rice, and lettuce) was 3.7 × 103 CFU/g without an enrichment step and 3.7 × 101 CFU/g following the 10-h enrichment. Hence, the assay developed here is specific and sensitive, providing an efficient tool for implementation in food for the simultaneous detection of B. cereus, L. monocytogenes, and S. aureus.

Donairs (Gyros) - Potential Hazards and Control

1986 ◽  
Vol 49 (5) ◽  
pp. 369-377 ◽  
Author(s):  
EWEN C. D. TODD ◽  
R. SZABO ◽  
F. SPIRING
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Heat Source ◽  
Bacillus Cereus ◽  
Clostridium Perfringens ◽  
Temperature Measurements ◽  
Microbiological Analysis ◽  
Short Period ◽  
And Control

Because of concerns that meat in donairs could allow growth of pathogens during cooking and overnight cooling of leftovers, 34 donairs from eleven establishments had temperatures taken and were examined microbiologically. Temperatures varied depending on depth of measurement and stage from the raw product to reheated leftovers. These were frequently >4 or <60°C and could be considered at temperatures favorable for growth of pathogens. Although aerobic colony counts were high (mean of 105 to 107 CFU/g), counts tended to decrease the longer the donair remained cooking on the spit. Staphylococcus aureus, Bacillus cereus, Clostridium perfringens and Escherichia coli were never more than 104/g despite some abusive practices, such as leaving donairs on the spit with the heat source turned off because the demand was low. Salmonella was found only in raw chicken slices to be used in donairs. It is recommended that good hygienic practices be encouraged at donair establishments and temperature measurements of donairs taken to verify these. Only if meat is <50°C at 1 cm below the surface during cooking or >5°C for the raw product or cooled leftovers, should samples be considered for microbiological analysis unless abusive practices have been observed. Because temperatures may vary over a short period of time during cooking, at least five measurements are recommended for each stage of the donair life (raw product, cooking donair, cooled leftovers and reheating donairs).

scholarly journals IDENTIFICATION OF FOOD PATHOGENS AND DETERMINATION OF THEIR DISTRIBUTION LEVEL IN UKRAINIAN FOOD PRODUCTS OF ANIMAL AND PLANT ORIGIN BY PCR METHOD

2019 ◽  
Vol 13 (4) ◽  
Author(s):  
O. Berhilevych ◽  
L. Pylypenko ◽  
V. Kasianchuk ◽  
A. Ilyeva ◽  
P. Shubin
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Bacillus Cereus ◽  
Clostridium Perfringens ◽  
Foodborne Pathogens ◽  
Shiga Toxin ◽  
Raw Materials ◽  
Cow Milk ◽  
Plant Origin ◽  
Pcr Method

The foodborne pathogens cause serious public health problems in each country. In this regard, microbiological investigation is included in food safety management of the food chain. Molecular methods and mostly polymerase chain reaction (PCR) are considered highly sensitive, specific and rapid methods for pathogens detection from raw material and food. This study describes the using of specially designed and highly specific primers for PCR to identify 5 common and especially dangerous causeve agents of food poisoning and disease and to determine their level of distribution in food of animal and plant origin. The studies included the identification of methicillin-resistant Staphylococcus aureus (MRSA) and Cronobacter spp. (E. sakazakii) from raw milk, Shiga toxin-producing strains of Escherichia coli (STEC) from beef and swine carcasses, Bacillus cereus and Clostridium perfringens from various types of plant and animal raw materials and products of its processing - fruits, vegetables, berries, dried and preserved products, food concentrates, half-canned food. A total of 397 food samples were investigated to detect these pathogens using classical bacteriological methods and PCR. It was found that the distribution of foodborne pathogens in the studied products of animal and plant origin was as follows: Staphylococcus aureus (MRSA) and Cronobacterspp. (E. sakazakii) in raw cow milk in 6.5% and 19.4% of cases, respectively; shiga-toxin-producing Escherichia coli (STEC) from beef and pork carcasses in 8.1% and 5.7%; Bacillus cereus and Clostridium perfringens from different types of plant and animal raw materials and their processing products averages 27.5 % and 7.7 %, respectively. The advantages of molecular biological methods to which the PCR method relates, include their speed, as well as the specificity of identification of microorganisms by the features of genetic regions of genes that carry information about their pathogenicity factors. It has been found that the rate of detection of these pathogens when using the PCR method in comparison with classical methods increases at least 5-9 times. This data will be useful for assessing microbiological risk and will help authorities develop strategies to reduce consumer health risks.

scholarly journals Association of Ct Values from Real-Time PCR with Culture in Microbiological Clearance Samples for Shiga Toxin-Producing Escherichia coli (STEC)

2020 ◽  
Vol 8 (11) ◽  
pp. 1801
Author(s):  
Michael Bording-Jorgensen ◽  
Brendon D. Parsons ◽  
Gillian A.M. Tarr ◽  
Binal Shah-Gandhi ◽  
Colin Lloyd ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Real Time ◽  
Shiga Toxin ◽  
Real Time Pcr ◽  
Gene Detection ◽  
Culture Positivity ◽  
Hands On ◽  
Culture Independent ◽  
Chromogenic Agar ◽  
Stool Samples

Shiga toxin-producing Escherichia coli (STEC) are associated with acute gastroenteritis worldwide, which induces a high economic burden on both healthcare and individuals. Culture-independent diagnostic tests (CIDT) in frontline microbiology laboratories have been implemented in Alberta since 2019. The objectives of this study were to determine the association between gene detection and culture positivity over time using STEC microbiological clearance samples and also to establish the frequency of specimen submission. Both stx genes’ amplification by real-time PCR was performed with DNA extracted from stool samples using the easyMAG system. Stools were inoculated onto chromogenic agar for culture. An association between gene detection and culture positivity was found to be independent of which stx gene was present. CIDT can provide rapid reporting with less hands-on time and technical expertise. However, culture is still important for surveillance and early cluster detection. In addition, stool submissions could be reduced from daily to every 3–5 days until a sample is negative by culture.

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