Advances in nucleic acid-based detection methods.

Wolcott, M J

doi:10.1128/cmr.5.4.370-386.1992

Cited by 5 publications

(6 citation statements)

References 44 publications

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“…DNA-based assays are becoming increasingly popular alternatives to serological and conventional methods for the detection of salmonellas in foods, animal feeds, carcasses, processing waters and other specimens because of their speed and potential specificity (Izat el al. 1989;Curiale et al 1990;Knight et al 1990;Wilson et al 1990;Bailey et al 1991 ;Hasan et al 1991 ;Rose et al 1991 ;Tsen et al 1991;Wang and Green, 1991;Widjojoatmodgjo et al 1991;Wolcott 1991;Can0 et al 1992b;Rahn et al 1992;Wolcott 1992). For the most part, however, DNA-based assays for the detection of salmonellas and other food pathogens require that each sample be processed and evaluated individually (see Wolcott 1991).…”

Section: Introductionmentioning

confidence: 99%

Fluorescent detection‐polymerase chain reaction (FD‐PCR) assay on microwell plates as a screening test for salmonellas in foods

Cano

Rasmussen

Fraga

et al. 1993

Journal of Applied Bacteriology

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This study evaluates a polymerase chain reaction assay coupled with a fluorescent detection in microwell plates for salmonellas in food samples. Chelex 100-extracted cultures and bulk and processed food samples were used as templates for a PCR assay in microwell plates, with a primer pair that amplifies a 206 bp segment of IS200. The PCR products were then denatured by heat and transferred to CovaLink NH plates (Nunc) to which capture oligonucleotides were covalently bound. Hybridization was performed for 1 h at 55 degrees C, the microwells were washed and an alkaline phosphatase-labelled probe, complementary of an internal sequence of the PCR product, was added. After stringent washes, 100 microliters of 1 mmol 1(-1) AttoPhos (JBL Scientific) was then added to the wells and the fluorescence measurement system (Millipore). The level of detection of the assay was as low as 1-10 cfu. A total of 172 food samples were tested, both by culture and FD-PCR. Of these 53 were culture positive and 119 culture negative. The sensitivity of the FD-PCR assay was 100% and the specificity was 90.1%. Positive and negative predictive values were 82.8 and 100%, respectively. Based on the results obtained in this study it appears that the FD-PCR assay described here can be useful to screen a large number of food samples for contamination by salmonellas.

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Section: Introductionmentioning

confidence: 99%

Fluorescent detection‐polymerase chain reaction (FD‐PCR) assay on microwell plates as a screening test for salmonellas in foods

Cano

Rasmussen

Fraga

et al. 1993

Journal of Applied Bacteriology

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“…However, they still have some limitations. It is not the purpose of this review to examine the strengths and weaknesses of these methods, as there are many excellent and recent reviews (Steffan and Atlas 1991;Wolcott 1992;Toze 1999;Strauba and Chandlerb 2003;Bankowski and Anderson 2004;Bustin and Nolan 2004;Mothershed and Whitney 2006;Karlen et al 2007;Lazcka et al 2007;Petti 2007) but to illustrate some of the limitations of the application of nuclei acid amplification methods to the direct detection of micro-organisms in water samples. The highly variable composition of target water samples, the generally low numbers of target micro-organisms in such samples and the physiological conditions of the target micro-organisms in this matrix largely determine the practicability of these methods.…”

Section: Discussionmentioning

confidence: 99%

Feasibility of methods based on nucleic acid amplification techniques to fulfil the requirements for microbiological analysis of water quality

Jofre

Blanch

2010

Journal of Applied Microbiology

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Summary Molecular methods based on nucleic acid recognition and amplification are valuable tools to complement and support water management decisions. At present, these decisions are mostly supported by the principle of end‐point monitoring for indicators and a small number of selected measured by traditional methods. Nucleic acid methods show enormous potential for identifying isolates from conventional culture methods, providing data on cultivable and noncultivable micro‐organisms, informing on the presence of pathogens in waters, determining the causes of waterborne outbreaks, and, in some cases, detecting emerging pathogens. However, some features of water microbiology affect the performance of nucleic acid‐based molecular techniques and thus challenge their suitability for routine water quality control. These features include the variable composition of target water samples, the generally low numbers of target micro‐organisms, the variable water quality required for different uses and the physiological status or condition of such micro‐organisms. The standardization of these molecular techniques is also an important challenge for its routine use in terms of accuracy (trueness and precision) and robustness (reproducibility and reliability during normal usage). Most of national and international water regulations recommend the application of standard methods, and any new technique must be validated respect to established methods and procedures. Moreover, molecular methods show a high cost‐effectiveness value that limits its practicability on some microbial water analyses. However, new molecular techniques could contribute with new information or at least to supplement the limitation of traditional culture‐based methods. Undoubtedly, challenges for these nucleic acid‐based methods need to be identified and solved to improve their feasibility for routine microbial water monitoring.

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“…and at 37°C for 20-24 h for the other species. Bacterial Polymerase chain reaction amplification (see Wolcott 1992) numbers were enumerated on BHI agar plates. All strains has been described for the detection of Y. enterocolitica (Wren were stored at −70°C in BHI containing 20% glycerol.…”

Section: Methodsmentioning

confidence: 99%

“…All strains has been described for the detection of Y. enterocolitica (Wren were stored at −70°C in BHI containing 20% glycerol. and Tabaqchali 1990;Fenwick and Murray 1991;Ibrahim et The number of indigenous bacteria in ground pork meat al. 1992;Nakajima et al 1992).…”

Section: Methodsmentioning

confidence: 99%

Detection of Yersinia enterocolitica in food by PCR amplification

Nilsson

Lambertz

Stålhandske

et al. 1998

Lett Appl Microbiol

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A polymerase chain reaction (PCR) assay was developed for detection of pathogenic, virulent strains of Yersinia enterocolitica. By using both virulence loci virF and ail as markers for pathogenicity, detection of species with a virulence factor present was possible. DNA preparation in the presence of hexadecyl trimethy ammonium bromide (CTAB) was followed by two 44 cycle amplification reactions, one for each of the markers. As few as 102Y. enterocolitica cells were detected in ground pork in the presence of 105–106 bacteria of other species. The described PCR assay provides a sensitive robust assay for the detection of virulent Y. enterocolitica in food.

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Advances in nucleic acid-based detection methods.

Cited by 5 publications

References 44 publications

Fluorescent detection‐polymerase chain reaction (FD‐PCR) assay on microwell plates as a screening test for salmonellas in foods

Fluorescent detection‐polymerase chain reaction (FD‐PCR) assay on microwell plates as a screening test for salmonellas in foods

Feasibility of methods based on nucleic acid amplification techniques to fulfil the requirements for microbiological analysis of water quality

Detection of Yersinia enterocolitica in food by PCR amplification

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