scholarly journals Enterotoxin-encoding genes in Staphylococcus aureus from buffalo milk

2019 ◽  
Vol 39 (8) ◽  
pp. 587-591
Author(s):  
Emmanuella O. Moura ◽  
Adriano H.N. Rangel ◽  
Cláudia S. Macêdo ◽  
Stela A. Urbano ◽  
Luciano P. Novaes ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Polymerase Chain Reaction ◽  
Dairy Products ◽  
Laboratory Tests ◽  
Animal Health ◽  
Food Poisoning ◽  
Buffalo Milk ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Encoding Genes

ABSTRACT: This paper investigated the occurrence of Staphylococcus aureus and the detection of enterotoxin-encoding genes of these strains in milk collected from 30 Murrah buffaloes used to produce dairy products in Brazil. A total of 68 strains of Staphylococcus aureus were found as identified by conventional laboratory tests, and thus screened for sea, seb, sec, sed, see, seg, seh and sei enterotoxin-encoding genes by polymerase chain reaction (PCR). Twelve strains containing enterotoxin-amplified genes were found, with higher expression for the sei and seh genes. These results can be attributed to animal health and inadequate cleaning of the equipment, indicating the need for better quality control in animal production and health lines. The results of this study with the presence of pathogens and their enterotoxigenic potential indicate a source of food poisoning, as well as being a pioneering study in the detection of new enterotoxins for buffalo milk.

Development of a Multiplex Polymerase Chain Reaction Assay for Detection and Differentiation of Staphylococcus aureus in Dairy Products†

2001 ◽  
Vol 64 (5) ◽  
pp. 664-668 ◽  
Author(s):  
SUDHIR TAMARAPU ◽  
JOHN L. McKILLIP ◽  
MARYANNE DRAKE
Keyword(s):  
Staphylococcus Aureus ◽  
Polymerase Chain Reaction ◽  
Multiplex Pcr ◽  
Dairy Products ◽  
Multiplex Polymerase Chain Reaction ◽  
Skim Milk ◽  
Cheddar Cheese ◽  
Polymorphism Analysis ◽  
Chain Reaction ◽  
Polymerase Chain

A multiplex polymerase chain reaction (PCR) assay was developed for the detection and differentiation of enterotoxigenic Staphylococcus aureus in dairy products. A solvent extraction procedure was successfully modified for extraction of S. aureus DNA from 10 ml of artificially contaminated skim milk or 20 g cheddar cheese. Primers targeting the enterotoxin C gene (entC) and thermostable nuclease gene (nuc) were used in the multiplex PCR. PCR products were confirmed using restriction fragment length polymorphism analysis. DNA was consistently quantified and amplified by uniplex PCR from 10 CFU/ml of S. aureus in skim milk or 10 CFU/20 g cheddar cheese. The sensitivity of the multiplex PCR was 100 CFU/ml of skim milk or 100 CFU/20 g cheddar cheese. The developed methodology allows presumptive identification and differentiation of enterotoxigenic S. aureus in less than 6 h.

scholarly journals Improved Multiplex Polymerase Chain Reaction for Rapid Staphylococcus Aureus Detection in Meat and Milk Matrices

2016 ◽  
Vol 15 (1) ◽  
pp. 65-76
Author(s):  
Zuzana Šramková ◽  
Barbora Vidová ◽  
Andrej Godány
Keyword(s):  
Staphylococcus Aureus ◽  
Polymerase Chain Reaction ◽  
Food Poisoning ◽  
Detection Sensitivity ◽  
23S Rrna ◽  
Rrna Gene ◽  
Chain Reaction ◽  
23S Rrna Gene ◽  
Polymerase Chain ◽  
Toxic Shock Syndrome Toxin

Abstract Staphylococcal food poisoning represents one of the most frequently occurring intoxications, caused by staphylococcal enterotoxins (SE-s) and staphylococcal enterotoxin-like proteins (SEl-s). Therefore, there is a need for rapid, sensitive and specific detection method for this human pathogen and its toxin genes in food matrices. The present work is focused on Staphylococcus aureus detection by a nonaplex polymerase chain reaction, which targets the 23S rRNA gene for identification of S. aureus at the species level, genes for classical SE-s (SEA, SEC, SED), new SE-s (SEH, SEI), SEl-s (SEK, SEL) and tsst-1 gene (toxic shock syndrome toxin). Primers were properly designed to avoid undesirable interactions and to create a reliably identifiable profile of amplicons when visualized in agarose gel. According to obtained results, this approach is able to reach the detection sensitivity of 12 colony forming units from milk and meat matrices without prior culturing and DNA extraction.

scholarly journals Molecular screening of Staphylococcus aureus enterotoxins associated with samples of meat / Iraq

2020 ◽  
Vol 11 (4) ◽  
pp. 6685-6691
Author(s):  
Sura I. A. Jabuk ◽  
Eman M. Jarallah
Keyword(s):  
Staphylococcus Aureus ◽  
Polymerase Chain Reaction ◽  
Molecular Screening ◽  
Chain Reaction ◽  
Local Markets ◽  
Fresh Frozen ◽  
Polymerase Chain ◽  
Staphylococcus Aureus Enterotoxins ◽  
Meat Samples ◽  
Encoding Genes

Staphylococcus aureus secreted many types of toxins accompanying Intestinal poisoning resulting from eating food contaminated with bacteria or their toxins. Five hundred meat samples were collected from local markets, including fresh, frozen, canned, sausage and hamburger to investigate their contamination with S.aureus and then determined their ability of these isolates to secrete enterotoxins by using polymerase chain reaction. The results showed that the ratio of isolated S.aureus is 30 (6%) and the percentage of encoding genes for toxins is 30(100%), 0(0%), 3(10), 0(0%),0(0%),3(10), 2(6.7), 1(3.3), 0(0%) and 3(10) to sea, seb, sec, sed, see, seg, see, sei, sej and sel respectively.The result shows the S.aureus isolated from contamination meat able to produce different type to enterotoxins sea, sec, seg, see, sei, and sel and present the sea toxin is the most prevalence type of staphylococcus enterotoxins. 

The Detection of Enterotoxins and Toxic Shock Syndrome Toxin Genes in Staphylococcus aureus by Polymerase Chain Reaction

2000 ◽  
Vol 63 (4) ◽  
pp. 479-488 ◽  
Author(s):  
J. McLAUCHLIN ◽  
G. L. NARAYANAN ◽  
V. MITHANI ◽  
G. O'NEILL
Keyword(s):  
Staphylococcus Aureus ◽  
Polymerase Chain Reaction ◽  
Toxic Shock Syndrome ◽  
Food Poisoning ◽  
Food Samples ◽  
Toxic Shock ◽  
Chain Reaction ◽  
Group I ◽  
Polymerase Chain ◽  
Toxic Shock Syndrome Toxin

A simple polymerase chain reaction (PCR)-based procedure was developed for the detection of fragments of staphylococcal enterotoxins (SEs) SEA, SEB, SEC, SED, SEE, SEG, SEH, and SEI together with the toxic shock syndrome toxin (TSST-1) genes of Staphylococcus aureus. One hundred and twenty-nine cultures of S. aureus were selected, 39 of which were recovered from 38 suspected staphylococcal food-poisoning incidents. The method was reproducible, and 32 different toxin genotypes were recognized. The presence of SE genes was associated with S. aureus strains reacting with phages in group III, and the TSST-1 gene with phages in group I. There was a 96% agreement between the PCR results for detection of SEA–D and TSST-1 as compared with a commercial reverse passive latex agglutination assay for the detection of SEs from cultures grown in vitro. Enterotoxin gene fragments were detected in S. aureus cultures recovered from 32 of the 38 suspected staphylococcal food poisoning incidents, and of these, 17 were associated with SEE, SEG, SEH, and SEI in the absence of SEA–D. Simple PCR procedures were also developed for the detection of SE directly in spiked food samples, and this was most successfully achieved in mushroom soup and ham. Detection was less successful in three types of cheese and in cream. SEA or SEB were detected by enzyme-linked immunosorbent assay in three food samples (two of which were associated with food poisoning incidents) naturally heavily contaminated with S. aureus: the appropriate SEA or SEB gene fragments were detected directly in these three foods by PCR.

Detection of Staphylococcus aureus in Dairy Products by Polymerase Chain Reaction Assay

2007 ◽  
Vol 6 (7) ◽  
pp. 857-862 ◽  
Author(s):  
Yang YANG ◽  
Xu-dong SU ◽  
Yao-wu YUAN ◽  
Chun-yu KANG ◽  
Ying-jun LI ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Assay ◽  
Dairy Products ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Molecular typing of Clostridium perfringens isolated from swine in slaughterhouses from São Paulo State, Brazil

2012 ◽  
Vol 42 (8) ◽  
pp. 1450-1456 ◽  
Author(s):  
Thais Sebastiana Porfida Ferreira ◽  
Andrea Micke Moreno ◽  
Renata Rodrigues de Almeida ◽  
Cleise Ribeiro Gomes ◽  
Debora Dirani Sena de Gobbi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Clostridium Perfringens ◽  
Food Poisoning ◽  
Positive Bacterium ◽  
Chain Reaction ◽  
High Prevalence ◽  
Fecal Isolate ◽  
Finishing Pig ◽  
Type C ◽  
Polymerase Chain

Clostridium perfringens is an anaerobic Gram-positive bacterium known as common pathogen for humans, for domestic and wildlife animals. Although infections caused by C. perfringens type C and A in swine are well studied, just a few reports describe the genetic relationship among strains in the epidemiological chain of swine clostridioses, as well as the presence of the microorganism in the slaughterhouses. The aim of the present study was to isolate C. perfringens from feces and carcasses from swine slaughterhouses, characterize the strains in relation to the presence of enterotoxin, alpha, beta, epsilon, iota and beta-2 toxins genes, using polymerase chain reaction (PCR) and comparing strains by means of Pulsed field gel electrophoresis (PFGE). Clostridium perfringens isolation frequencies in carcasses and finishing pig intestines were of 58.8% in both types of samples. According to the polymerase chain reaction assay, only alfa toxin was detected, being all isolates also negative to enterotoxin and beta2 toxin. Through PFGE technique, the strains were characterized in 35 pulsotypes. In only one pulsotype, the isolate from carcass sample was grouped with fecal isolate of the same animal, suggesting that the risk of cross-contamination was low. Despite the high prevalence of C. perfringens in swine carcasses from the slaughterhouses assessed, the risk of food poisoning to Brazilian pork consumers is low, since all strains were negative to cpe-gene, codifying enterotoxin.

scholarly journals Pertussis in north-central and northwestern regions of Algeria

2016 ◽  
Vol 10 (11) ◽  
pp. 1191-1199 ◽  
Author(s):  
Nabila Benamrouche ◽  
Hassiba Tali Maamar ◽  
Malika Lazri ◽  
Sonia Hasnaoui ◽  
Abdelkarim Radoui ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Vaccination Coverage ◽  
Laboratory Tests ◽  
Chain Reaction ◽  
North Central ◽  
Household Contacts ◽  
Source Of Infection ◽  
Antimicrobial Sensitivity ◽  
Adolescents And Adults ◽  
Polymerase Chain

Introduction: Pertussis outbreaks continue to occur in many countries despite high vaccination coverage. Under-diagnosed cases in adolescents and adults may result in increased transmission to infants, who are at risk of severe pertussis. Additional measures to protect both groups should be considered. Methodology: Nasopharyngeal samples and sera were collected from patients and household contacts with clinically suspected pertussis. Diagnoses were confirmed by culture, real-time polymerase chain reaction (PCR), and serology. Bordetella pertussis isolates were characterized by antimicrobial sensitivity and fimbrial serotyping. Results: Of 392 participants, 134/248 patients (54%) and 66/144 contacts (45.8%) had confirmed pertussis infections. B. parapertussis was not detected. All B. pertussis isolates were sensitive to the antibiotics tested, and all expressed the Fim3, not the Fim2, fimbrial serotype. Most patients (81.2%) were <6 months (51.8% of whom were <3 months) of age; 77.6% were unvaccinated, and most positive contacts were mothers 20–40 years of age. Conclusions: Despite high vaccination coverage, pertussis is circulating in Algeria. Most infections occur in unvaccinated infants <6 months of age, with mothers as the main source of infection. An adolescent/adult booster should be considered. Adoption of sensitive and specific laboratory tests would improve pertussis diagnosis and surveillance.

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