A survey on the incidence of Campylobacter spp. and the development of a surface adhesion polymerase chain reaction (SA-PCR) assay for the detection of Campylobacter jejuni in retail meat products

2001 ◽  
Vol 18 (3) ◽  
pp. 287-298 ◽  
Author(s):  
Orla M. Cloak ◽  
Geraldine Duffy ◽  
J.J. Sheridan ◽  
I.S. Blair ◽  
D.A. McDowell
Keyword(s):  
Polymerase Chain Reaction ◽  
Campylobacter Jejuni ◽  
Meat Products ◽  
Surface Adhesion ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Retail Meat ◽  
Polymerase Chain ◽  
Campylobacter Spp

scholarly journals Collaborative Trial for Validation of a Real-Time Reverse Transcriptase-Polymerase Chain Reaction Assay for Detection of Central Nervous System Tissues as Bovine Spongiform Encephalopathy RiskMaterial: Part 1

2006 ◽  
Vol 89 (5) ◽  
pp. 1335-1340
Author(s):  
Amir Abdulmawjood ◽  
Holger Schnenbrcher ◽  
Michael BÜlte
Keyword(s):  
Central Nervous System ◽  
Polymerase Chain Reaction ◽  
Real Time ◽  
Meat Products ◽  
Spongiform Encephalopathy ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Heat Treated ◽  
Polymerase Chain

Abstract A collaborative trial was conducted to evaluate a real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assay for detection of central nervous system (CNS) tissues in meat products (e.g., sausages). The method is based on the detection of ruminant glial fibrillary acidic protein (GFAP) mRNA by applying real-time RT-PCR. The assay was evaluated through a multicenter trial involving 12 participating laboratories that received coded cDNA obtained from 3 different types of sausages. The participants used 5 different real-time detection systems. The results obtained in this validation revealed that this real-time RT-PCR assay performed well in the different laboratories with a detection limit of at least 0.1% CNS in those test materials that contained strongly heat-treated samples (sausages cooked at 120C) and the medium heat-treated samples (sausages cooked at 80C). The detection limit of liver sausages was determined to be 0.2% of CNS. Neither the samples with no CNS additive nor the bovine DNA and the negative control containing 100% swine brain gave any positive signals. The presented results indicate that the real-time RT-PCR assay was just as reproducible between laboratories, as repeatable within a laboratory, could reliably be used for detection of bovine spongiform encephalopathy risk material in meat and meat products, and signify that it may be used with confidence in any laboratory.

scholarly journals Survey of chicken abattoir for the presence of Campylobacter jejuni and Campylobacter coli

2006 ◽  
Vol 48 (6) ◽  
pp. 307-310 ◽  
Author(s):  
Ana L.L. Cortez ◽  
Angela C.F.B. Carvalho ◽  
Eliana Scarcelli ◽  
Simone Miyashiro ◽  
Ana M.C. Vidal-Martins ◽  
...  
Keyword(s):  
Public Health ◽  
Polymerase Chain Reaction ◽  
Campylobacter Jejuni ◽  
Intestinal Tract ◽  
Campylobacter Coli ◽  
Improve Quality ◽  
Chain Reaction ◽  
Paulo State ◽  
Polymerase Chain ◽  
Campylobacter Spp

The genus Campylobacter is of great importance to public health because it includes several species that may cause diarrhea. These species may be found in water, food and in the intestinal tract of chickens. This study investigated the presence of Campylobacter jejuni and Campylobacter coli in chicken abattoirs in São Paulo State, Brazil. A total of 288 samples of feces, feathers, scald water, evisceration water, chiller water, and the rinse water of eviscerated, not eviscerated and chilled carcasses were collected in six chicken abattoirs. Polymerase Chain Reaction (PCR) was performed in Campylobacter spp.-positive isolates using the gene HIP, specific for hippuricase enzyme from Campylobacter jejuni and aspartokinase gene, specific to detect Campylobacter coli. The percentage of positive isolates of Campylobacter jejuni was 4.9% (14/288). Isolation was greater in feces samples (22%, 8/36). One sample was positive for the species C. coli. In conclusion, the results indicate that it is necessary to improve quality control for Campylobacter spp. in chicken abattoirs.

Detection of Thermophilic Campylobacter spp. in Blood-Free Enriched Samples of Inoculated Foods by the Polymerase Chain Reaction

2000 ◽  
Vol 63 (3) ◽  
pp. 299-303 ◽  
Author(s):  
RICHARD L. THUNBERG ◽  
TONY T. TRAN ◽  
MARK O. WALDERHAUG
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Extraction ◽  
Raw Milk ◽  
Extraction Techniques ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Enrichment Broth ◽  
Thermophilic Campylobacter ◽  
Polymerase Chain ◽  
Campylobacter Spp

The detection of thermophilic Campylobacter spp., as represented by Campylobacter jejuni, by the polymerase chain reaction (PCR) was investigated and compared with the selective agar isolation (SAI) method. Stationary-phase cultures of C. jejuni were inoculated into either blood-free enrichment broth (BFEB) or BFEB that contained 10% broccoli, crabmeat, mushroom, raw milk, and raw oyster rinses. Following a 48-h enrichment period, aliquots of food test portions were removed for simultaneous analysis by PCR and SAI. It was determined that the presence of charcoal and iron in the enrichment broth interfered with the PCR assay. Therefore, three DNA extraction techniques were developed and evaluated using a 16S rRNA primer pair in the PCR assay. The 50% end point (DL50) values (determined upon six initial C. jejuni spiking levels) were used to assess the frequency of isolation utilizing PCR versus SAI for the detection of this organism in the enrichment matrices. There were virtually no differences in detection of C. jejuni among enriched samples analyzed by PCR and SAI. Mean DL50 values (n = 3) for plain BFEB, broccoli, crabmeat, mushroom, raw milk, and raw oyster were, respectively, 0.02 (PCR) versus 0.01 (SAI), 0.01 versus 0.06, 0.07 versus 0.04, 0.03 versus 0.08, 0.01 versus 0.01, and 0.01 versus 0.01 CFU/5 g food. Significant variability in the detection limit of C. jejuni by PCR in the food enrichments was observed among DNA extraction techniques. Using 48-h enrichment cultures followed by PCR analysis could save 1 day of the time required for the presumptive identification of C. jejuni in suspected foods.

Polymerase Chain Reaction Detection of Listeria monocytogenes on Frankfurters Using Oligonucleotide Primers Targeting the Genes Encoding Internalin AB

2003 ◽  
Vol 66 (2) ◽  
pp. 237-241 ◽  
Author(s):  
YONG SOO JUNG ◽  
JOSEPH F. FRANK ◽  
ROBERT E. BRACKETT ◽  
JINRU CHEN
Keyword(s):  
Polymerase Chain Reaction ◽  
Listeria Monocytogenes ◽  
Meat Products ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Oligonucleotide Primers ◽  
Pcr Product ◽  
Genes Encoding ◽  
Pure Cell ◽  
Polymerase Chain

A polymerase chain reaction (PCR) assay targeting the genes encoding internalin AB (inlAB) was developed for detecting Listeria monocytogenes in pure cell cultures and on artificially contaminated frankfurters. Four sets of oligonucleotide primers were evaluated. The set targeting a 902-bp region of the inlAB gene was the most specific. This PCR product was detected in 51 L. monocytogenes strains belonging to four different serogroups (1/2a, 1/2b, 1/2c, and 4b). In contrast, the PCR product was not detected in other Listeria spp. (Listeria innocua, Listeria ivanovii, Listeria seeligeri, Listeria welshimeri, or Listeria grayi) or in gram-positive, non-Listeria bacteria, indicating that the primer set was highly specific for L. monocytogenes. The detection limit of the PCR assay was 105 CFU per ml of pure cell culture. However, the assay could detect as few as 101 CFU of L. monocytogenes in 25 g of frankfurter with 16 h of enrichment in modified Listeria enrichment broth at 30°C. The total assay time including enrichment was approximately 24 h. These results suggest that the PCR assay can be used to rapidly detect L. monocytogenes on frankfurters and possibly other types of ready-to-eat meat products.

Identification of Campylobacter jejuni Isolates from Cloacal and Carcass Swabs of Chickens in Thailand by a 5′Nuclease Fluorogenic Polymerase Chain Reaction Assay

2002 ◽  
Vol 65 (11) ◽  
pp. 1712-1716 ◽  
Author(s):  
PAWIN PADUNGTOD ◽  
ROBERT HANSON ◽  
DAVID L. WILSON ◽  
JULIA BELL ◽  
JOHN E. LINZ ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Campylobacter Jejuni ◽  
Rrna Genes ◽  
23S Rrna ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Gyra Gene ◽  
Test Kit ◽  
Polymerase Chain ◽  
Conventional Test

A rapid 5′ nuclease fluorogenic polymerase chain reaction (PCR) assay for identifying Campylobacter jejuni was applied to Campylobacter isolates from chicken cloacal and carcass swabs collected from three chicken farms and a slaughterhouse in Thailand. The primers and the probe were based on the sequence of the gyrA gene in C. jejuni. C. jejuni isolates were identified by fluorogenic PCR assay of bacterial cells directly from Campylobacter-selective agar medium. This assay allowed the identification of C. jejuni within 1 day after colonies appeared on selective media. The fluorogenic PCR assay yielded results comparable to those of the conventional test kit (kappa = 0.76) but required less time. When the two methods disagreed with regard to species identification, results were confirmed by PCR restriction fragment length polymorphism of 23S rRNA genes. In these instances, the fluorogenic PCR assay correctly identified more isolates of C. jejuni than did the conventional test kit (six of seven isolates were unidentifiable by the conventional test kit). The fluorogenic PCR assay is a rapid and specific method that outperforms the conventional test kit in the identification of C. jejuni from environmental samples.

Detection of Mink (Mustela vison) DNA in Meat Products using Polymerase Chain Reaction PCR Assay

2012 ◽  
Vol 7 (12) ◽  
pp. 1348-1355 ◽  
Author(s):  
Weili Sun ◽  
Guangyu Li ◽  
Hanlu Liu ◽  
Wei Zhong ◽  
Haihua Zhang ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Meat Products ◽  
Mustela Vison ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Polymerase Chain

Rapid Detection of Campylobacter jejuni in Chicken Rinse Water by Melting-Peak Analysis of Amplicons in Real-Time Polymerase Chain Reaction

2003 ◽  
Vol 66 (8) ◽  
pp. 1343-1352 ◽  
Author(s):  
ZHIHUI CHENG ◽  
MANSEL W. GRIFFITHS
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Dna Extraction ◽  
Campylobacter Jejuni ◽  
Real Time Pcr ◽  
Campylobacter Coli ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Polymerase Chain ◽  
Triton X

Five DNA extraction protocols for the detection of Campylobacter spp. by polymerase chain reaction (PCR) were compared. A method involving Triton X-100 produced template DNA of sufficient quality to allow the detection of Campylobacter jejuni at levels of 100 CFU/ml in pure culture. Primers were designed on the basis of the cadF gene sequence. With a SYBR Green I real-time PCR assay, these primers amplified only sequences present in C. jejuni to produce a product with a melting temperature of 81.5°C. None of the strains of Campylobacter coli, Campylobacter lari, or Campylobacter fetus tested produced this product during the PCR assay. Other noncampylobacter species tested were shown not to possess the cadF sequence. The real-time PCR combined with a rapid, simple Triton X-100 DNA extraction protocol made it possible to detect <10 CFU of C. jejuni per ml of chicken rinse within 14 h.

Polymerase chain reaction amplification and typing of rotavirus in environmental samples

1995 ◽  
Vol 31 (5-6) ◽  
pp. 371-374 ◽  
Author(s):  
R. Gajardo ◽  
R. M. Pintó ◽  
A. Bosch
Keyword(s):  
Polymerase Chain Reaction ◽  
Environmental Samples ◽  
Polymerase Chain Reaction Amplification ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Group A ◽  
Reaction Amplification ◽  
Stool Specimens ◽  
Polymerase Chain

A reverse transcription polymerase chain reaction (RT-PCR) assay is described that has been developed for the detection and serotyping of group A rotavirus in stool specimens and concentrated and non-concentrated sewage specimens.

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