Development and evaluation of aptamer magnetic capture assay in conjunction with real-time PCR for detection of Campylobacter jejuni

LWT ◽  
2014 ◽  
Vol 56 (2) ◽  
pp. 256-260 ◽  
Author(s):  
Soo Hwan Suh ◽  
Hari P. Dwivedi ◽  
Lee-Ann Jaykus
Keyword(s):  
Real Time ◽  
Campylobacter Jejuni ◽  
Real Time Pcr ◽  
Capture Assay ◽  
Magnetic Capture

Schnellnachweis von Campylobacter jejuni und Campylobacter coli aus Lebensmittelvoranreicherungen mittels Real-Time-PCR sowie immunomagnetischer Separation

2005 ◽  
Vol 67 (03) ◽  
Author(s):  
A Mayr ◽  
H Söllner ◽  
R Zucker ◽  
A Wieland ◽  
A Notzon ◽  
...  
Keyword(s):  
Real Time ◽  
Campylobacter Jejuni ◽  
Real Time Pcr ◽  
Campylobacter Coli

Quantitative Detection of Campylobacter jejuni on Fresh Chicken Carcasses by Real-Time PCR

2007 ◽  
Vol 70 (6) ◽  
pp. 1373-1378 ◽  
Author(s):  
ANNA-CLARA RÖNNER ◽  
HANS LINDMARK
Keyword(s):  
Detection Limit ◽  
Real Time ◽  
Dna Extraction ◽  
Campylobacter Jejuni ◽  
Real Time Pcr ◽  
Close Correlation ◽  
Quantitative Detection ◽  
Quantitative Real Time Pcr ◽  
Direct Plating ◽  
Pcr Method

Campylobacter jejuni infection is a significant cause of foodborne gastroenteritis worldwide. Consumption and handling of poultry products is believed to be the primary risk factor for campylobacteriosis. Risk assessments require quantitative data, and C. jejuni is enumerated usually by direct plating, which sometimes allows growth of non-Campylobacter bacteria. The objective of the present study was to develop a quantitative real-time PCR method (q-PCR) for enumerating C. jejuni in chicken rinse without a culturing step. The procedure to obtain the template for the PCR assay involved (i) filtration of 10 ml of chicken rinse, (ii) centrifugation of the sample, and (iii) total DNA extraction from the pellet obtained using a commercial DNA extraction kit. The detection limit of the method was comparable to that for plating 100 μl of chicken rinse on modified charcoal cefoperazone deoxycholate agar, and the detection limit could be further improved 10-fold by concentrating the DNA eluate by ethanol precipitation. A close correlation for spiked chicken rinse was obtained for the results of the quantitative real-time PCR method and direct plating (r = 0.99). The coefficient of correlation for the methods was 0.87 when samples from chicken carcasses on the slaughter line were analyzed, whereas a lower correlation (r = 0.76) was obtained when samples from retail carcasses were analyzed. Greater variation in the proportion of dead and/or viable but not culturable Campylobacter types in the retail samples may explain the decreased correlation between the methods. Overall, the new method is simple and fast and the results obtained are closely correlated with those for direct plating for samples containing a low proportion of dead Campylobacter cells.

scholarly journals Detection and quantification of Campylobacter in foods: New analytic approaches to detect and quantify Campylobacter spp. in food samples

2020 ◽  
Vol 9 (2) ◽  
Author(s):  
Maria Francesca Peruzy ◽  
Yolande Thérèse Rose Proroga ◽  
Federico Capuano ◽  
Federica Corrado ◽  
Serena Santonicola ◽  
...  
Keyword(s):  
Real Time ◽  
Campylobacter Jejuni ◽  
Internal Control ◽  
Real Time Pcr ◽  
Quantitative Methods ◽  
Bacterial Load ◽  
Digital Pcr ◽  
Food Samples ◽  
Meat Samples ◽  
Campylobacter Spp

The aim of the present study was to develop rapid qualitative and quantitative methods based on the use of Real-Time PCR and Droplet Digital PCR (ddPCR), in order to have reliable techniques to detect and quantify Campylobacter spp. in food samples. The gene 16S-rRNA was used as specific target for Campylobacter spp. Real- Time PCR evaluation assay and a not competitive internal control was ushered in it. To investigate the selectivity of the method, 26 Campylobacter strains and 40 non-Campylobacter strains were tested and in order to verify the application of Real-Time PCR method, 5 pork meat samples were experimentally inoculated with a Campylobacter jejuni strain. Subsequently, dilutions with a bacterial load of Campylobacter jejuni within 10-106 CFU/mL were chosen for the optimization of the ddPCR assay. Lastly, a total of 54 naturally contaminated foods samples were analyzed through molecular (Real-Time PCR and ddPCR) and traditional methods. The Real-Time PCR protocol demonstrated to amplify only the Campylobacter spp. strains and when Campylobacter jejuni was experimentally inoculated in meat samples the pathogen was always detected. The ddPCRs assay allowed to quantify a level of contamination of 10 CFU/mL, but it was unable to quantify levels of 105 – 106 CFU/mL. Lastly, Campylobacter spp. was never detected in the 54 samples tested. In conclusion, the novel analytic approach proposed, based on an initial screening of the samples with Real-Time PCR and then on quantification of Campylobacter spp. with a ddPCR on those positive, represents a quick monitoring tool and, if used correctly, it would allow the implementation of food safety.

A more sensitive, efficient and ISO 17025 validated Magnetic Capture real time PCR method for the detection of archetypal Toxoplasma gondii strains in meat

2017 ◽  
Vol 47 (13) ◽  
pp. 875-884 ◽  
Author(s):  
Ignacio Gisbert Algaba ◽  
Manon Geerts ◽  
Malgorzata Jennes ◽  
Wim Coucke ◽  
Marieke Opsteegh ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Real Time ◽  
Real Time Pcr ◽  
Iso 17025 ◽  
Magnetic Capture ◽  
Pcr Method

scholarly journals A semi-automated magnetic capture probe based DNA extraction and real-time PCR method applied in the Swedish surveillance of Echinococcus multilocularis in red fox (Vulpes vulpes) faecal samples

2014 ◽  
Vol 7 (1) ◽  
Author(s):  
Mats Isaksson ◽  
Åsa Hagström ◽  
Maria Teresa Armua-Fernandez ◽  
Helene Wahlström ◽  
Erik Olof Ågren ◽  
...  
Keyword(s):  
Real Time ◽  
Dna Extraction ◽  
Real Time Pcr ◽  
Echinococcus Multilocularis ◽  
Vulpes Vulpes ◽  
Red Fox ◽  
Capture Probe ◽  
Magnetic Capture ◽  
Faecal Samples ◽  
Pcr Method

Brain is the predilection site of Toxoplasma gondii in experimentally inoculated pigs as revealed by magnetic capture and real-time PCR

2014 ◽  
Vol 38 ◽  
pp. 167-170 ◽  
Author(s):  
Jana Juránková ◽  
Walter Basso ◽  
Helena Neumayerová ◽  
Vojtech Baláž ◽  
Eva Jánová ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Real Time ◽  
Real Time Pcr ◽  
Magnetic Capture ◽  
Predilection Site

Real-time PCR assay for rapid detection and quantification of Campylobacter jejuni on chicken rinses from poultry processing plant

2007 ◽  
Vol 21 (3) ◽  
pp. 177-181 ◽  
Author(s):  
Aradom Debretsion ◽  
Tsegaye Habtemariam ◽  
Saul Wilson ◽  
David Nganwa ◽  
Teshome Yehualaeshet
Keyword(s):  
Real Time ◽  
Campylobacter Jejuni ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
Processing Plant ◽  
Pcr Assay ◽  
Poultry Processing ◽  
Detection And Quantification

Rapid detection and differentiation of pathogenic Campylobacter jejuni and Campylobacter coli by real-time PCR

2005 ◽  
Vol 156 (1) ◽  
pp. 107-114 ◽  
Author(s):  
Marwan Abu-Halaweh ◽  
J. Bates ◽  
Bharat K.C. Patel
Keyword(s):  
Real Time ◽  
Campylobacter Jejuni ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
Campylobacter Coli
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