Evaluation of a Cytolethal Distending Toxin (cdt) Gene‐Based Species‐Specific Multiplex PCR Assay for the Identification of Campylobacter Strains Isolated from Poultry in Thailand

Samosornsuk, Worada; Asakura, Masahiro; Yoshida, Emi; Taguchi, Takashi; Nishimura, Kazuhiko; Eampokalap, Boonchuay; Phongsisay, Vongsavanh; Chaicumpa, Wanpen; Yamasaki, Shinji

doi:10.1111/j.1348-0421.2007.tb03974.x

Cited by 36 publications

(45 citation statements)

References 35 publications

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“…Briefly, heat-denatured genomic DNA (300 ng) was applied to a GeneScreen Plus (DuPont, Boston, MA, USA). The membranes were hybridized with cdt gene probes under highly stringent conditions as described previously (18). The cdtA, cdtB, and cdtC gene probes for C. jejuni, C. coli, and C. fetus were amplified by PCR using the primer sets described above.…”

Section: Methodsmentioning

confidence: 99%

“…PCR for hipO gene detection and 16S rRNA gene amplification was performed as described previously (17,18). The PCR primers and conditions used in this study are summarized in Table 1.…”

Section: Methodsmentioning

confidence: 99%

“…Images were captured on a Bio-Rad Gel Doc system (Bio-Rad Laboratories, Inc.). DNA preparation: Bacterial chromosomal DNA was purified and evaluated with respect to quantity and quality as described previously (18).…”

Section: Methodsmentioning

confidence: 99%

“…Sequencing of 16S rRNA gene: 16S rRNA gene sequence analysis was performed as described previously (18).…”

Section: Methodsmentioning

confidence: 99%

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Isolation and Characterization of Campylobacter Strains from Diarrheal Patients in Central and Suburban Bangkok, Thailand

et al. 2015

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SUMMARY: Campylobacter-induced diarrhea is increasingly recognized worldwide. However, little information is available regarding the Campylobacter strains associated with diarrheal patients in Thailand. In this study, we attempted to isolate Campylobacter strains from diarrheal patients in Thailand and to characterize the species using a cytolethal distending toxin (cdt) gene-based C. jejuni, C. coli, and C. fetus-specific multiplex PCR assay. Campylobacter species were also confirmed using 16S rRNA gene sequencing and hipO gene detection. From 2,500 diarrheal stool specimens, 76 Campylobacter-like organisms were isolated and identified via conventional culture methods. Among these 76 organisms, 73 were identified as Campylobacter species (43 C. jejuni, 29 C. coli, and 1 C. fetus) via multiplex PCR, whereas 3 remained unidentified. Two Campylobacter-like organisms yielded 2 amplicons corresponding to cdt genes from C. jejuni and C. coli. Subsequently, C. jejuni and C. coli were reisolated from each sample. The third isolate was identified as C. hyointestinalis via 16S rRNA gene sequencing. To our knowledge, this is the first report on the isolation of C. hyointestinalis from a diarrheal patient in Thailand. These data indicate that C. jejuni (58z) and C. coli (40z) are prevalent among diarrheal patients in Thailand.

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Section: Methodsmentioning

confidence: 99%

“…PCR for hipO gene detection and 16S rRNA gene amplification was performed as described previously (17,18). The PCR primers and conditions used in this study are summarized in Table 1.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Sequencing of 16S rRNA gene: 16S rRNA gene sequence analysis was performed as described previously (18).…”

Section: Methodsmentioning

confidence: 99%

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Isolation and Characterization of Campylobacter Strains from Diarrheal Patients in Central and Suburban Bangkok, Thailand

et al. 2015

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“…Asakura et al [2] observed that cdt genes were universally distributed in C. jejuni, C. coli and C. fetus in a species-specific manner. Based on this finding, a cdt genebased species-specific multiplex PCR targeting either cdtA, cdtB, or cdtC gene has been developed [3] and evaluated for the accuracy of the species identification with a large number of isolates of C. jejuni, C. coli and C. fetus of clinical or animal origin [14,30].…”

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confidence: 99%

A Cytolethal Distending Toxin Gene-Based Multiplex PCR Assay for Detection of Campylobacter spp. in Stool Specimens and Comparison with Culture Method

Shiramaru

Asakura

Inoue

et al. 2012

The Journal of Veterinary Medical Science

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ABSTRACT. In this study, we evaluated the applicability of cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR for the direct detection and identification of Campylobacter jejuni, C. coli and C. fetus from stool specimens of patients with gastroenteritis in comparison to culture methods. A total of 711 stool specimens were examined for the isolation or detection of campylobacters by using Skirrow's selective agar culture plates, a filtration method and the multiplex PCR assay. Forty-one and 36 C. jejuni strains were isolated by culture and filtration methods, respectively. In addition, 2 and 3 C. coli strains were isolated by Skirrow and the filtration methods, respectively. However, when the multiplex PCR was employed, the cdtB genes of C. jejuni and C. coli were detected in 45 and 4 stool samples, respectively, and 9 C. jejuni PCR-positive samples by multiplex PCR were negative by culture method. Sequence analysis of the PCR products obtained from 8 stool specimens from which campylobacters were not isolated by culture method but the sequences exactly matched with that of the cdtB gene of C. jejuni strain 81-176. None of the remaining stool samples which were culture negative for campylobacters produced any amplicon. Stool samples were defined as Campylobacterpositive if detected by any method. The sensitivity of the multiplex PCR was 83%, which was higher than Skirrow (74%) and filtration method (66%). These data indicate that cdtB gene-based multiplex PCR is a rapid and more sensitive method to identify the most important species of Campylobacter for human diseases. (248)

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Investigation of Campylobacter fetus in breeding bulls of private farms in Bangladesh

Hoque

Islam

Saddam

et al. 2022

Veterinary Medicine & Sci

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Background Bovine genital campylobacteriosis (BGC) is a venereal disease caused by Campylobacter fetus that has a negative impact on animal reproduction. The bull is considered to be a symptomless carrier that spreads the disease agent to breeding cows, causing infertility and sporadic abortion. Aim The study aims to estimate the prevalence, identify risk factors of Campylobacter fetus ( C. fetus ) infection and antimicrobial resistance pattern of the C. fetus isolates. Method A cross‐sectional survey was conducted in Mymensingh district of Bangladesh. Bull smegma samples (single sample from each bull) were collected from 300 bulls from four farms and tested via culture, biochemical identification and finally 16S rRNA and cdtA gene‐based molecular assays (PCR) for herd and animal‐level prevalence estimation. Herd‐ and animal‐level data on risk factors were collected from the farmers using a pretested questionnaire and analysed by univariable and multivariable logistic regression models with a p value of <0.05 was taken statistically significant for both analyses. Results Among the surveyed farms, 75% (95% CI: 19.4%–99.4%) were confirmed to have bulls infected with Campylobacter fetus at herd level. However, animal‐level occurrence of C. fetus was estimated to be 8.7% (26/300) (95% CI: 5.7%–12.4%). Natural service increases the odds of campylobacteriosis 38.18 times (95% CI: 13.89–104.94) in comparison to artificial insemination for C . fetus infection in bulls. Significantly, half of the isolates (50%, n = 13) were identified to be multidrug resistant (MDR) for three to five antimicrobial agents. Conclusion This study highlights the need to develop official guidelines for C. fetus control and prevention in Bangladesh including mandatory artificial insemination in reproductive cows and heifers, routine screening of breeding bulls for C. fetus free status.

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Evaluation of a Cytolethal Distending Toxin (cdt) Gene‐Based Species‐Specific Multiplex PCR Assay for the Identification of Campylobacter Strains Isolated from Poultry in Thailand

Cited by 36 publications

References 35 publications

Isolation and Characterization of <i>Campylobacter</i> Strains from Diarrheal Patients in Central and Suburban Bangkok, Thailand

Isolation and Characterization of <i>Campylobacter</i> Strains from Diarrheal Patients in Central and Suburban Bangkok, Thailand

A Cytolethal Distending Toxin Gene-Based Multiplex PCR Assay for Detection of <i>Campylobacter</i> spp. in Stool Specimens and Comparison with Culture Method

Investigation of Campylobacter fetus in breeding bulls of private farms in Bangladesh

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