Development of a Single-Reaction Multiplex PCR Toxin Typing Assay for
            <i>Staphylococcus aureus</i>
            Strains

Sharma, Naresh Kumar; Rees, Catherine; Dodd, C.E.R.

doi:10.1128/aem.66.4.1347-1353.2000

Cited by 123 publications

(90 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The surveillance of staphylococcal contamination within foods has been applied globally as critical tactic in public health. Sharma et al (35) reported that 41% of isolated S. aureus from milk bulk tank were capable of producing enterotoxin. In our investigation, only about 10% of staphylococcal isolates were positive for SEA.…”

Section: Discussionmentioning

confidence: 99%

Enterotoxin A Gene Barrier Staphylococcus aureus Within Traditionally Dairy Products of Tehran

Hassani¹,

Doust²,

Mobarez³

2014

Int J Enteric Pathog

View full text Add to dashboard Cite

Background: Staphylococcus aureusis a serious agent that often colonize dairy products all over the world. Staphylococcal enterotoxins are the essential causes of food poisoningin human societies. Enterotoxin type A is an importantstaphylococcal exotoxin. Objectives: The aim of present study was to detect the enterotoxin producing Staphylococcus.aureus within different dairy products collected from Tehran, Iran. Materials and Methods: Two hundreds twenty dairy products samples were collected from local dealers across the city. The samples were first screened for S. aureus contaminations. All isolated strains of S. aureus were then investigated for enterotoxin a gene, usind spesific primer sets. Results: Staphylococcus aureus was isolated from 43% of dairy samples: 22% from milk and 18% from cheese samples. The SEA genes were detected in 10 isolates (22%) originated from raw milk and in two isolates (25%) from domestic cheese. Conclusions: Since, the staphylococcal enterotoxins are heat stable, heat had no effect on the toxicity of the enterotoxins within positive samples. Our primer stets confirmed previous studies that introduced PCR as rapid, sensitive, and specific method for dairy products screening system. Our data showed that routine screening and surveillance is vital for different food materials including dairy products.

show abstract

Section: Discussionmentioning

confidence: 99%

Enterotoxin A Gene Barrier Staphylococcus aureus Within Traditionally Dairy Products of Tehran

Hassani¹,

Doust²,

Mobarez³

2014

Int J Enteric Pathog

View full text Add to dashboard Cite

show abstract

“…More recently, methods for S. aureus toxin typing by multiplex PCR have been reported (6,26,29). These PCR methods are based on combinations of ent gene-specific primers or a combination of universal forward primers and specific reverse primers (36).…”

Section: Discussionmentioning

confidence: 99%

“…Other techniques have been used to identify toxin genotypes, including DNA-DNA hybridization and PCR, but these protocols were designed to detect only one or a few toxin genes (21,35). Multiplex PCR for detection of several ent genes has been reported (6,26,29,30,36), but additional restriction endonuclease assays or other steps are required to ensure unambiguous identification of ent-specific amplicons. Therefore, there is still a need for a rapid and specific method for simultaneous detection and identification of SEs for diagnostic and epidemiological purposes.…”

mentioning

confidence: 99%

Simultaneous Analysis of Multiple Staphylococcal Enterotoxin Genes by an Oligonucleotide Microarray Assay

et al. 2004

View full text Add to dashboard Cite

Staphylococcal enterotoxins (SEs) are a family of 17 major serological types of heat-stable enterotoxins that are one of the leading causes of gastroenteritis resulting from consumption of contaminated food. SEs are considered potential bioweapons. Many Staphylococcus aureus isolates contain multiple SEs. Because of the large number of SEs, serological typing and PCR typing are laborious and time-consuming. Furthermore, serological typing may not always be practical because of antigenic similarities among enterotoxins. We report on a microarray-based one-tube assay for the simultaneous detection and identification (genetic typing) of multiple enterotoxin (ent) genes. The proposed typing method is based on PCR amplification of the target region of the ent genes with degenerate primers, followed by characterization of the PCR products by microchip hybridization with oligonucleotide probes specific for each ent gene. We verified the performance of this method by using several other techniques, including PCR amplification with gene-specific primers, followed by gel electrophoresis or microarray hybridization, and sequencing of the enterotoxin genes. The assay was evaluated by analysis of previously characterized staphylococcal isolates containing 16 ent genes. The microarray assay revealed that some of these isolates contained additional previously undetected ent genes. The use of degenerate primers allows the simultaneous amplification and identification of as many as nine different ent genes in one S. aureus strain. The results of this study demonstrate the usefulness of the oligonucleotide microarray assay for the analysis of multitoxigenic strains, which are common among S. aureus strains, and for the analysis of microbial pathogens in general.

show abstract

“…Para la identificación de las cepas de S. aureus por PCR, se amplificó una secuencia del gen de la nucleasa termoestable (nuc), con los cebadores descritos por primera vez por Brakstad, et al (9) (cuadro 1). Los oligonucleótidos cebadores empleados para identificar los genes de enterotoxinas (sea-see) fueron los propuestos por Sharma, et al (10). Este diseño permite, en una sola reacción de amplificación, la detección de genes que codifican estas enterotoxinas.…”

Section: Materiales Y Métodosunclassified

Estrategia de PCR múltiple para la caracterización molecular simultánea de Staphylococcus aureus y enterotoxinas estafilocócicas en aislamientos de brotes de origen alimentario

Brizzio¹,

Tedeschi

Zalazar

2012

biomedica

View full text Add to dashboard Cite

Development of a Single-Reaction Multiplex PCR Toxin Typing Assay for Staphylococcus aureus Strains

Cited by 123 publications

References 49 publications

Enterotoxin A Gene Barrier Staphylococcus aureus Within Traditionally Dairy Products of Tehran

Enterotoxin A Gene Barrier Staphylococcus aureus Within Traditionally Dairy Products of Tehran

Simultaneous Analysis of Multiple Staphylococcal Enterotoxin Genes by an Oligonucleotide Microarray Assay

Estrategia de PCR múltiple para la caracterización molecular simultánea de Staphylococcus aureus y enterotoxinas estafilocócicas en aislamientos de brotes de origen alimentario

Contact Info

Product

Resources

About