Rapid and simultaneous analysis of five foodborne pathogenic bacteria using multiplex PCR

Guan, Zhuo; Jiang, Yun; Gao, Feng; Lin, Zhang; Zhou, Guang Hong; Guan, Zheng Jun

doi:10.1007/s00217-013-2039-1

Cited by 39 publications

(13 citation statements)

References 33 publications

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“…In the last 10 years, many authors have proposed the use of PCR for the detection of foodborne pathogens to replace the time consuming culture based classical techniques. They are rapid, easy to handle, sensitive and specific and therefore constitute very valuable tools for routine applications.The result of optimization for validation of PCR on different gradient annealing temperatures from 56˚C to 63 ˚C on foodborne pathogens concerned in this study show great accordance with the results of (Chen et al, 2012); (Thapa et al, 2013)and (Kim et al, 2014) where they used gradient PCR on validation and detection of S. aureus, L. monocytogenes and E.coli but with different primers used in this study while (Guan et al, 2013) and (latha et al, 2014)used gradient PCR on validation and detection of S. aureus and L. monocytogenes with same primers used in this study.On the other hand, (Kupradit et al, 2013)used gradient PCR on validation and detection ofL. monocytogenes and E. coli with only the (uspA) gene of E. coli with the prfA gene of L. monocytogenes.…”

Section: Discussionsupporting

confidence: 82%

Detection of some foodborne pathogens in meat products by Polymerase Chain Reaction

Armany

Ahmed

Ibrahim

et al. 2016

Benha Veterinary Medical Journal

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A total of 100 random samples of meat products including raw minced meat, raw sausage, luncheon and basterma (25 samples of each) were collected from different markets in Cairo and Giza governorates to be examined bacteriologically for detection of Staphylococcus aureus, Listeria monocytogenes and Escherichia coli. These samples were examined for isolation of such pathogens by conventional bacteriological methods and by polymerase chain reaction (PCR). Concerning S.aureus bacteriological results revealed the prevalence in minced meat, Sausage, luncheon and basterma was (24%, 24%, 20%, 4%) respectively. While L. monocytogenes revealed the prevalence in minced meat, sausage, luncheon and basterma was (4%, 0%, 0%, 0%) respectively and E. coli revealed the prevalence in minced meat, sausage, luncheon and basterma was (20%, 20%, 24%, 20%) respectively. The results cleared that PCR is an ideal method for identification of foodborne pathogens, as it was effective, less labor, more sensitive, reduces effort and time after using gradient PCR in validation of each microbe.

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Section: Discussionsupporting

confidence: 82%

Detection of some foodborne pathogens in meat products by Polymerase Chain Reaction

Armany

Ahmed

Ibrahim

et al. 2016

Benha Veterinary Medical Journal

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“…As the hly gene is unique to L. monocytogenes , it is most commonly used for detection using PCR or qPCR (Guan et al . ). Supplemental in silico analysis of amplicon specificity was carried out by using nucleotide blast (http://blast.ncbi.nlm.nih.gov/).…”

Section: Methodsmentioning

confidence: 97%

“…The highly specific genomic regions chosen for selective identification of bacteria included Listeriolysin O encoding hly gene from the L. monocytogenes (EU372057) genome and the Shiga toxin 2 encoding stx gene from E. coli O157:H7 (AB048837) genome. As the hly gene is unique to L. monocytogenes, it is most commonly used for detection using PCR or qPCR (Guan et al 2013). Supplemental in silico analysis of amplicon specificity was carried out by using nucleotide BLAST (http://blast.ncbi.nlm.nih.gov/).…”

Section: Qpcr Primers and Probe Designmentioning

confidence: 99%

A duplex qPCR for the simultaneous detection of Escherichia coli O157:H7 and Listeria monocytogenes using LNA probes

Nitecki

Teape²,

Carney

et al. 2015

Lett Appl Microbiol

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The study highlights a novel duplex qPCR for Listeria monocytogenes and Escherichia coli O157:H7 that could be used as an alternative to plate-based ISO or singleplex PCR methods while minimizing the costs. The assay uses rapid DNA extraction methods and locked nucleic acid probes. Sensitivity and specificity are 100 and 98·95% respectively. The potential for quantitative rage of the assay is 10(8) -10(1) CFU ml(-1) .

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“…Proteinase K is to digest proteins including membrane proteins, Sodium acetate can be utilized to precipitate high molecular weight molecules including genomic DNA. The successive treatment with 70% ethanol allows an additional purification, or wash, of the nucleic acid from the remaining [13,2,21,25,26] generally result in high efficiency DNA extractions, this material it effect the efficiently of PCR, according to [12], he limit residual concentration it should be less than SDS 0.005%, Phenol 0.2%, Ethanol 1%, Isopropanol 1%, Sodium acetate 5 mM, Sodium chloride 25 mM, EDTA 0.5 mM. However, the phenolic/ethanol procedure is time consuming and relates to the use of harmful organic chemicals.…”

Section: Phenol/ Ethanol Methodsmentioning

confidence: 99%

“…Despite improvements in meat processing hygiene practices in recent years, the occurrence of foodborne pathogenic microorganisms is still commonplace. The absolute requirement for safe meat highlights the need for a rapid and accurate identification of these foodborne pathogens [25]. Electrophoreses and nanodrop showed that all the DNA extraction methods were successfully from artificial contamination, and different quantity depend on the initial portion used, it was measured by using nanodrop.…”

Section: Artificial Contamination Samplesmentioning

confidence: 99%

Comparative Study of Rapid DNA Extraction Methods of Pathogenic Bacteria

Abdelhai¹

2016

BIO

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Abstract:Detection of pathogenic bacteria in food is most important for food safety and quality control, and the critical step it chooses the rapid, sensitive and more economical method to extract DNA to produce high quality and decrease the timeconsuming of measuring. Extraction of nucleic acids is the first step in most molecular biology studies and in all recombinant DNA techniques, but the difficult access steps and critical of analysis. Here we report, describe and compare the simple and fast methods of extraction (physical, boiling, phenol/ethanol and commercial kit) methods, from pure culture and then from beef samples. The quantity and quality of extraction methods were confirmed by polymerase chain reaction, agarose gel electrophoresis, and spectrophotometer nanodrop. Results revealed that the efficiently for all three methods were significant compared with the commercial kit, however, in pure culture the boiling method sex tract its more efficient, convenient and cheaper method for template preparation and significant when it compare with other methods while in beef samples experimental results showed that the phenol/ethanol method extract its more significantly.

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Rapid and simultaneous analysis of five foodborne pathogenic bacteria using multiplex PCR

Cited by 39 publications

References 33 publications

Detection of some foodborne pathogens in meat products by Polymerase Chain Reaction

Detection of some foodborne pathogens in meat products by Polymerase Chain Reaction

A duplex qPCR for the simultaneous detection of Escherichia coli O157:H7 and Listeria monocytogenes using LNA probes

Comparative Study of Rapid DNA Extraction Methods of Pathogenic Bacteria

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