Specificity of the BAX Polymerase Chain Reaction System for Detection of the Foodborne Pathogen Listeria monocytogenes

Stewart, Diana; Gendel, Steven M.

doi:10.1093/jaoac/81.4.817

Cited by 29 publications

(11 citation statements)

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“…1994). Stewart and Gendel (1998) reported that the detection sensitivity obtained with the BAX" protease was slightly less than that obtained with the treatment with lysozyme and proteinase K. In the present study, higher concentrations of iron in the undiluted broth may have inhibited the PCR reactions by enhancing protease degradation of the Taq polymerase. It was subsequently determined in the present study that prior cell lysis was not required for successful PCR and the BAX" protease actually inhibited PCR.…”

Section: Discussioncontrasting

confidence: 53%

SIMPLE oPSU BROTH‐BAX® PCR‐PICOGREEN® SYSTEM FOR RAPID DETECTION OF LISTERIA MONOCYTOGENES IN PASTEURIZED MILK AND HOT DOGS

Parameswaran¹,

Guyer

Knabel

2003

Journal of Food Safety

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Factors that significantly affect BAX® PCR detection of Listeria monocytogenes from optimized Penn State University (oPSU) broth were identified and optimized. Concentration of PCR product was significantly reduced by BAX™ protease and significantly increased by eliminating the lysis step and directly diluting oPSU broth prior to PCR. A simple oPSU broth‐BAX® PCR‐PicoGreen® (PSU‐BAX‐PicoGreen) system was developed and compared with current methods for detecting low levels of L. monocytogenes in commercial milk and hot dogs. All 30 milk samples inoculated with 10–20 CFU per mL L. monocytogenes were positive by FDA, BAX® and PSU‐BAX‐PicoGreen methods and all 42 uninoculated milk samples were negative by all of the above methods. All 30 hot dog samples inoculated with 10‐20 CFU/g L. monocytogenes were positive by the USDA and PSU‐BAX‐PicoGreen methods, however, 2 hot dog samples gave indeterminate results with the standard BAX® method. All 42 uninoculated hot dog samples were negative by USDA, 9 were indeterminate by BAX® and 2 were positive by PSU‐BAX‐PicoGreen. The PSU‐BAX‐PicoGreen system may provide a simple and accurate method for rapidly screening pasteurized foods for both injured and noninjured L. monocytogenes.

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Section: Discussioncontrasting

confidence: 53%

SIMPLE oPSU BROTH‐BAX® PCR‐PICOGREEN® SYSTEM FOR RAPID DETECTION OF LISTERIA MONOCYTOGENES IN PASTEURIZED MILK AND HOT DOGS

Parameswaran¹,

Guyer

Knabel

2003

Journal of Food Safety

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“…The use of an internal standard as a positive control for the PCR assay for bacterial detection has been described by several authors (Bansal 1996;Monteiro et al 1997;Bailey 1998;Steward and Gendel 1998;Fach et al 1999;Rijpens et al 1999;Straub et al 1999). This internal control (IC) was useful for detecting false-negative results caused by the presence of compounds in the DNA extract which inhibit DNA ampli®cation and, consequently, for determining the bacterial contamination of a sample.…”

Section: Discussionmentioning

confidence: 99%

Campylobacter contamination in French chicken production from farm to consumers. Use of a PCR assay for detection and identification of Campylobacter jejuni and Camp. coli

et al. 2001

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Aims: Campylobacter contamination in French chicken production from the farm to the consumer was determined using a PCR assay for bacteria detection and identi®cation. Methods and Results: Samples were bird droppings from poultry houses, neck skins, livers, hearts, gizzards, wings, legs and escalopes from slaughterhouses and gizzards, legs, drumstick, breast and escalopes from a supermarket. Bacterial DNA extraction was performed after an enrichment step in a broth and was followed by PCR. An internal control (IC) was used for both DNA extraction and PCR. Campylobacter were detected in 79á2% of poultry houses. Of the 303 samples, 201 were Campylobacter-positive (i.e. 66á3%) including 43á2% faecal samples, 5á6% slaughterhouse samples and 17á5% supermarket samples. There was no signi®cant difference between the molecular method and the conventional culture technique for Campylobacter detection whatever the samples. The sensitivity was 5 UFC g)1 of samples and 1á5´10 3 UFC ml )1 of enrichment broth. The use of IC revealed PCR inhibition in 13 samples and problems in the DNA extraction in ®ve samples. Conclusions: Signi®cant Campylobacter contamination affects all stages of French chicken production. Signi®cance and Impact of the Study: The understanding of Campylobacter contamination at different levels of chicken production and the determination of the best place(s) for intervention are important for signi®cantly decreasing Campylobacteriosis. Our technique is rapid and can be used on different chicken samples for Campylobacter detection and identi®cation.

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“…It is used to increase sensitivity and specificity and has been used for the identification of L. monocytogenes in clinical samples [57,[107][108][109][110], environmental samples [78], and milk samples [97,111,112]. Commercially available PCR tests are the BAXÒ PCR system (Qualicon, Wilmington, DE, USA) and the ProbeliaÒ assay (Sanofi Diagnostic Pasteur, Marne la Coquette, France), which have been trialed in the field on a variety of different sample types [113][114][115][116][117][118][119][120][121][122].…”

Section: Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

Methods for the isolation and identification ofListeriaspp. andListeria monocytogenes: a review

Hughes²,

2005

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Listeria monocytogenes is an important food-borne pathogen and is widely tested for in food, environmental and clinical samples. Identification traditionally involved culture methods based on selective enrichment and plating followed by the characterization of Listeria spp. based on colony morphology, sugar fermentation and haemolytic properties. These methods are the gold standard; but they are lengthy and may not be suitable for testing of foods with short shelf lives. As a result more rapid tests were developed based on antibodies (ELISA) or molecular techniques (PCR or DNA hybridization). While these tests possess equal sensitivity, they are rapid and allow testing to be completed within 48 h. More recently, molecular methods were developed that target RNA rather than DNA, such as RT-PCR, real time PCR or nucleic acid based sequence amplification (NASBA). These tests not only provide a measure of cell viability but they can also be used for quantitative analysis. In addition, a variety of tests are available for sub-species characterization, which are particularly useful in epidemiological investigations. Early typing methods differentiated isolates based on phenotypic markers, such as multilocus enzyme electrophoresis, phage typing and serotyping. These phenotypic typing methods are being replaced by molecular tests, which reflect genetic relationships between isolates and are more accurate. These new methods are currently mainly used in research but their considerable potential for routine testing in the future cannot be overlooked.

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Specificity of the BAX Polymerase Chain Reaction System for Detection of the Foodborne Pathogen Listeria monocytogenes

Cited by 29 publications

References 0 publications

SIMPLE oPSU BROTH‐BAX® PCR‐PICOGREEN® SYSTEM FOR RAPID DETECTION OF LISTERIA MONOCYTOGENES IN PASTEURIZED MILK AND HOT DOGS

SIMPLE oPSU BROTH‐BAX® PCR‐PICOGREEN® SYSTEM FOR RAPID DETECTION OF LISTERIA MONOCYTOGENES IN PASTEURIZED MILK AND HOT DOGS

Campylobacter contamination in French chicken production from farm to consumers. Use of a PCR assay for detection and identification of Campylobacter jejuni and Camp. coli

Methods for the isolation and identification ofListeriaspp. andListeria monocytogenes: a review

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