Specific identification, grouping and differentiation of Campylobacter jejuni among thermophilic campylobacters using multiplex PCR

Korolik, Victoria; Friendship, D. T.; Peduru-Hewa, T.; Alfredson, David; Fry, Benjamin N.; Coloe, Peter J.

doi:10.1017/s0950268801005763

Cited by 12 publications

(9 citation statements)

References 19 publications

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“…Heat lysis techniques have been successfully applied for the isolation of template DNA from diagnostic specimens and thus offer considerable time and labor savings for the routine application of DNA-based diagnostics (29,36,40,45,61). Although crude sample processing does not remove all potential PCRinhibitory substances, 5Ј Taq nuclease assays appear to be more robust, enabling successful amplification of target material as demonstrated in this study.…”

Section: Discussionmentioning

confidence: 90%

Comparison of Culture and a Novel 5′ Taq Nuclease Assay for Direct Detection of Campylobacter fetus subsp. venerealis in Clinical Specimens from Cattle

McMillen¹,

Fordyce²,

Doogan³

et al. 2006

J Clin Microbiol

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A Campylobacter fetus subsp. venerealis-specific 5 Taq nuclease PCR assay using a 3 minor groove binder-DNA probe (TaqMan MGB) was developed based on a subspecies-specific fragment of unknown identity (S. Hum, K. Quinn, J. Brunner, and S. L. On, Aust. Vet. J. 75:827-831, 1997). The assay specifically detected four C. fetus subsp. venerealis strains with no observed cross-reaction with C. fetus subsp. fetus-related Campylobacter species or other bovine venereal microflora. The 5 Taq nuclease assay detected approximately one single cell compared to 100 and 10 cells in the conventional PCR assay and 2,500 and 25,000 cells from selective culture from inoculated smegma and mucus, respectively. The respective detection limits following the enrichments from smegma and mucus were 5,000 and 50 cells/inoculum for the conventional PCR compared to 500 and 50 cells/inoculum for the 5 Taq nuclease assay. Field sampling confirmed the sensitivity and the specificity of the 5 Taq nuclease assay by detecting an additional 40 bulls that were not detected by culture. Urine-inoculated samples demonstrated comparable detection of C. fetus subsp. venerealis by both culture and the 5 Taq nuclease assay; however, urine was found to be less effective than smegma for bull sampling. Three infected bulls were tested repetitively to compare sampling tools, and the bull rasper proved to be the most suitable, as evidenced by the improved ease of specimen collection and the consistent detection of higher levels of C. fetus subsp. venerealis. The 5 Taq nuclease assay demonstrates a statistically significant association with culture ( 2 ‫؍‬ 29.8; P < 0.001) and significant improvements for the detection of C. fetus subsp. venerealis-infected animals from crude clinical extracts following prolonged transport.

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Section: Discussionmentioning

confidence: 90%

Comparison of Culture and a Novel 5′ Taq Nuclease Assay for Direct Detection of Campylobacter fetus subsp. venerealis in Clinical Specimens from Cattle

McMillen¹,

Fordyce²,

Doogan³

et al. 2006

J Clin Microbiol

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“…The plates were incubated for 48–72 h at 42°C in a 90% N 2 /5% CO 2 /5% O 2 atmosphere. Colonies with Campylobacter ‐like morphology were Gram stained, and their identity was confirmed by PCR analysis as required (Korolik et al , 2001).…”

Section: Methodsmentioning

confidence: 99%

“…These models are also used to examine the behavior of isogenic mutants in vivo and cloned genes with potential pathogenicity characteristics (Nachamkin et al, 1993;Wassenaar et al, 1993;Purdy et al, 1999;Ziprin et al, 1999;Fry et al, 2000). There are essentially two types of chicken colonization model: one using very young chickens, 0-2-days-old, and one using 14-16-day-old chickens (Caw-thraw et al, 1996;Rice et al, 1997;Korolik et al, 1998Korolik et al, , 2001Chen & Stern, 2001;Ringoir & Korolik, 2003). In this study, we compare the two types of model in terms of infectious dose of the C. jejuni cells required to colonize the chickens of different ages, and the susceptibility of chickens of different ages to colonization by various C. jejuni strains.…”

Section: Introductionmentioning

confidence: 99%

Comparison of 2-day-old and 14-day-old chicken colonization models forCampylobacter jejuni

Ringoir

Szylo

Korolik

2007

FEMS Immunol Med Microbiol

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In this study, we compared two types of chicken infection models for Campylobacter jejuni in terms of infectious dose required to colonize the chickens and the susceptibility of chickens of different ages to persistent colonization by C. jejuni. In one model, chickens at day 2 posthatching were used, and in the other, 14-day-old chickens were used. The minimum C. jejuni cell number required to colonize 14-day-old chickens was 5 Â 10 4 cells, and that for 2-day-old chickens was 5 Â 10 3 . The ability of various C. jejuni strains to colonize the chicken gastrointestinal tract was the same in both models.

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“…Based on studies in experimentally infected chicken flocks, it has been proposed that some C. jejuni strains are capable of becoming dominant in flocks infected with different genotypes (Barrow and Page 2000; Korolik et al. 2001).…”

Section: Discussionmentioning

confidence: 99%

Genetic variability of Campylobacter jejuni isolated from fresh and frozen broiler carcasses

et al. 2006

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Aims: The aim of this study was to determine the genetic variability of Campylobacter jejuni isolates from poultry before and after freezing treatment in order to identify genotypes that would survive the treatment. Methods and Results: C. jejuni was isolated from both fresh and frozen halves of the same carcass after freezing for 2 or more than 20 days at −20°C. From 36 carcasses, representing five unrelated flocks in Norway, a total of 209 isolates were included in the study. Thirty‐two of the isolates were recovered with a qualitative method while the remaining 177 were isolated using a quantitative method. Isolates were genotyped with fluorescent amplified fragment length polymorphism using MfeI and BglII restriction enzymes. Nine different genotypes were identified, however, one genotype was shown to be dominant in three different flocks. This genotype and the dominant genotype of another flock were found among isolates from fresh and frozen broiler halves. They were also shown to be identical to genotypes frequently identified among strains isolated from humans, cattle and poultry flocks in previous years. Conclusions: Freezing treatment or isolation method appeared not to select for a particular genotype. Significance and Impact of the Study: The results of the present study indicate that the freezing tolerance of strains is not genotype dependent.

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Specific identification, grouping and differentiation of Campylobacter jejuni among thermophilic campylobacters using multiplex PCR

Cited by 12 publications

References 19 publications

Comparison of Culture and a Novel 5′ Taq Nuclease Assay for Direct Detection of Campylobacter fetus subsp. venerealis in Clinical Specimens from Cattle

Comparison of Culture and a Novel 5′ Taq Nuclease Assay for Direct Detection of Campylobacter fetus subsp. venerealis in Clinical Specimens from Cattle

Comparison of 2-day-old and 14-day-old chicken colonization models forCampylobacter jejuni

Genetic variability of Campylobacter jejuni isolated from fresh and frozen broiler carcasses

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