An alternative method for Staphylococcus aureus DNA isolation

Chapaval, L.; Moon, David Henry; Gomes, José Elias; Duarte, Fernanda Oliveira; Tsai, Siu Mui

doi:10.1590/s0102-09352008000200004

Cited by 23 publications

(15 citation statements)

References 16 publications

(11 reference statements)

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“…Since S.aureus does not compete with indigenous microorganisms within raw milk and dairy products, staphylococcal contamination is usually associated with improper handling of food materials, followed by storage under poor environmental conditions (12). Staphylococcus aureus is also present in animals; dairy cattle (10), sheep, and goats, particularly if they are affected by subclinical mastitis, are likely the source of milk contamination (11).…”

Section: Introductionmentioning

confidence: 99%

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

Hassani¹,

Doust²,

Mobarez³

2014

Int J Enteric Pathog

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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.

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Section: Introductionmentioning

confidence: 99%

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

Hassani¹,

Doust²,

Mobarez³

2014

Int J Enteric Pathog

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“…They designed primers specific for the nuc gene (NUC1-NUC2 for S. aureus, NUC3-NUC4 for S. hyicus and NUC5-NUC6 for S. intermedius) and detected three target species by MPCR, directly from bovine whole milk, with adequate specificity and acceptable detection limit for identification of coagulase positive Staphylococcus (CPS) in foods with the detection limit of 10 3 CFU/ml which is in contrast to our study as we could not multiplex One plausible reason might be the cell wall of gram positive bacteria, that contains vide variety of molecules that serve to provide rigid exoskeleton for protection against both mechanical and osmotic lysis. It has been observed that Staphylococcus aureus detection from milk by molecular techniques was reported when lysostaphin a peptidase produced by Streptococcus simulans that break the cellular membrane of Staphylococcus aureus was used in DNA extraction protocol (Chapaval et al, 2008). We too attribute it due to the difference between extraction of DNA from Gram positive and Gram negative or it may be possible that template DNA of Staphylococcus required more sophisticated approach which we did not apply because we wanted to use multiplex directly on field samples.…”

Section: Evaluation Of Mpcr On Field Samplesmentioning

confidence: 98%

Development of a multiplex PCR for identification of mastitis causing organisms

Singh

Chandra

Kaur

et al. 2019

IJDS

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In the present study, a de novo multiplex polymerase chain reaction using genus specific primers for four organisms' viz., E. coli, Klebsiella pneumoniae, Staphylococcus aureus and Streptococcus agalactiae were designed to identify and differentiate all the four organisms in a single reaction. Further, DNA was extracted from mastitic milk using three different methods and was subjected to multiplex polymerase chain reaction to compare and identify the best method out of these three methods based on the amplification results of multiplex polymerase chain reaction. It was observed from the study that multiplex PCR was able to amplify three organisms (Streptococcus agalactiae, Klebsiella pneumoniae and E. coli) successfully whereas was unable to amplify Staphylococcus aureus in the multiplex PCR. Mutiplex PCR when evaluated on the field samples using mastitic milk revealed SDS Triton DNA extraction method to be superior to Power Food Microbial DNA isolation Kit and Milk Bacterial DNA isolation kit method in isolating the bacterial DNA from the mastitic milk as indicated by amplification in multiplex PCR reaction.

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“…Various protocols have been described for the extraction of DNA from specific groups of microorganisms; however, the efficiency of these protocols varies among different groups (Baratto and Megiolaro, 2012;Chapaval et al, 2008;Ligozzi and Fontana, 2003;Rivera et al, 2003;Wilson, 1997). This variation is mainly due to the inherent characteristics of the different bacterial groups and the structure of their cell walls, which reflects the efficiency of lysis.…”

Section: Choice Of a Single Protocol For Gram Positive And Negative Bmentioning

confidence: 99%

Choice of DNA extraction protocols from Gram negative and positive bacteria and directly from the soil

Tiago¹,

Naiara²,

Shana³

et al. 2015

Afr. J. Microbiol. Res.

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DNA extraction is a fundamentally important step for the implementation of genotypic techniques in microbial identification, and the use of such techniques has become essential for the analysis of soil microbial diversity. Considering culture independent methodologies, it is still necessary to ensure that DNA is extracted in appropriate amounts and that extracted DNA is inhibitor-free. This study aimed at selecting a single protocol suitable for the extraction of total DNA from Gram positive and negative bacteria isolated from different sources, as well as a protocol for the direct extraction of DNA from soil. Four experimental protocols and a commercial kit were tested for the extraction of total DNA from isolated bacteria. Among the protocols, the detergent + salt + thermal incubation method (based on Harju et al., 2004) was considered the most promising because it produced satisfactory yields of DNA, with adequate quality for all isolates studied, especially Staphylococcus aureus, without the need to use enzymes and glass beads which can make the extraction process more expensive. Three experimental protocols and the commercial kit were tested for the direct extraction of DNA from soil. Regarding PCR amplification, the amount of total DNA extracted is less limiting than its quality. Thus, commercial kit PowerMax™ Soil DNA Isolation (MoBio) offered more promising results, because although this provided low yields of DNA, it was sufficient for polymerase chain reaction (PCR) amplification.

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An alternative method for Staphylococcus aureus DNA isolation

Cited by 23 publications

References 16 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

Development of a multiplex PCR for identification of mastitis causing organisms

Choice of DNA extraction protocols from Gram negative and positive bacteria and directly from the soil

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