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http://dx.doi.org/10.17555/jvc.2015.04.32.2.154

Development of a Panel of Multiplex Real-Time Polymerase Chain Reaction Assays for Simultaneous Detection of Canine Enteric Bacterial Pathogens  

Jang, Hye-Jin (Veterinary Laboratory Medicine, College of Veterinary Medicine, Chungbuk National University)
Han, Jae-Ik (Veterinary Internal Medicine, College of Veterinary Medicine, Konkuk University)
Kang, Hyo-Min (Veterinary Laboratory Medicine, College of Veterinary Medicine, Chungbuk National University)
Na, Ki-Jeong (Veterinary Laboratory Medicine, College of Veterinary Medicine, Chungbuk National University)
Publication Information
Journal of Veterinary Clinics / v.32, no.2, 2015 , pp. 154-157 More about this Journal
Abstract
A major cause of diarrhea in a dog is an infection with bacteria which include Salmonella spp., Campylobacter (C.) spp., and Clostridium (Cl.) spp.. It is fastidious to identify these bacteria by the culture. The purpose of this experiment is to devise the method for detecting Cl. perfringens, C. jejuni, C. coli, and Salmonella spp. with rapid and high sensitivity. The fecal samples collected from 71 normal and 66 diarrheic dog feces were used to compare the prevalence of the enteric pathogens and to develop a multiplex real-time polymerase chain reaction (PCR) assay for clinical use. Detection of Cl. perfringens, C. coli, and C. jejuni in diarrhea feces was higher than normal feces. A developed multiplex real-time PCR is useful for determining the presence and quantity of pathogen-specific or other unique sequences with in a fecal sample.
Keywords
PCR; Clostridium perfringens; Campylobacter jejuni; Campylobacter coli; Salmonella spp.;
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1 Amani J, Mirhosseini SA, lmani Fooladi AA. A review approaches to identify enteric bacterial pathogens. Jundishapur J Microbiol 2015; 8: e17473.
2 Belanger SD, Boissinot M, Clairoux N, Picard FJ, Bergeron MG. Rapid detection of Clostridium difficile in Feces by real-time PCR. J Clin Microbiol 2003; 41: 730-734.   DOI
3 Bochelmann U, Domes HH, Ayuso-Gabella MN, Salgot de Marcay M, Tandoi V, Levantesi C, Masciopinto C, Van Houtte E, Szewzyk U, Wintqens T, Grohmann E. Quantitative PCR monitoring of antibiotic resistance genes and bacterial pathogens in three European artificial groundwater recharge systems. Appl Environ Microbiol 2009; 75: 154-163.   DOI
4 Russello G, Russo A, Sisto F, Scaltrito MM, Farina C. Laboratory diagnosis of Clostridium difficile associated diarrhoea and molecular characterization of clinical isolates. New Microbiol 2012; 35: 307-316.
5 Skanseng B, Kaldhusdal M, Rudi K. Comparison of chicken gut colonisation by the pathogens Campylobacter jejuni and Clostridium perfringens by real-time quantitative PCR. Mol Cell Probes 2006; 20: 269-279.   DOI
6 Volkmann H, Schwartz T, Bischoff P, Kirchen S, Obst U. Detection of clinically relevant antibiotic-resistance genes in municipal wastewater using real-time PCR (TaqMan). J Microbiol Methods 2004; 56: 277-286.   DOI
7 Liu J, Gratz J, Amour C, Kibiki G, Becker S, Janaki L, Verweij JJ, Taniuchi M, Sobuz SU, Haque R, Haverstick DM, Houpt ER. A laboratory developed Taqman Array Card for simultaneous detection of nineteen enteropathogens. J Clin Microbiol 2013; 5: 472-480.
8 Chaban B, Ngeleka M, Hill JE. Detection and quantification of 14 Campylobacter species in pet dogs reveals and increase in species richness in feces of diarrheic animals. BMC Microbiol 2010; 10: 1-7.   DOI
9 Jihong L, Vicki Adams, Trudi L. Bannam, Kazuaki M, Jorge P, Francisco A, Julian I, Bruce A. Toxin Plasmids of Clostridium perfringens. Microbiol Mol Biol Rev 2015; 79: 193-224.   DOI
10 Jokinen CC, Koot JM, Carrillo CD, Gannon VP, Jardine CM, Mutschall SK, Topp E, Taboada EN. An enhanced technique combining pre-enrichment and passive filtration increases the isolation efficiency of Campylobacter jejuni and Campylobacter coli from water and animal fecal samples. J Microbiol Methods 2012; 91: 506-513   DOI
11 Maddocks S, Olma T, Chen S. Comparison of CHROMagar Salmonella medium and xylose-lysine-desoxycholate and Salmonella-Shigella agars for isolation of Salmonella strains from stool samples. J Clin Microbiol 2002; 40: 2999-3003.   DOI
12 Marks SL, Rankin SC, Byrne BA, Weese JS. Enteropathogenic Bacteria in Dogs and Cats: Diagnosis, Epidemiology, Treatment, and Control. J Vet Intern Med 2011; 25: 1195-1208.   DOI
13 Meer RR, Songer JG. Multiplex polymerase chain reaction assay for genotyping Clostridium perfringens. Am J Vet Res 1997; 58: 702-5.
14 Mokhtari W, Nsaibia S, Gharbi A, Aouni M. Real-time PCR using SYBR Green for the detection of Shigella spp. in food and stool samples. Mol Cell Probes 2013; 27: 53-59.   DOI
15 Rinttila T, Lyra A, Kroqius-Kurikka L, Palva A. Real-time PCR analysis of enteric pathogens from fecal samples of irritable bowel syndrome subjects. Gut Pathog 2011; 3: 6-14.   DOI
16 Daniel Paredes-Sabja, Sarker MR. Germination response of spores of the pathogenic bacterium Clostridiun perfringens and Clostridium difficile to cultured human epithelial cells. Anaerobe 2011; 17: 78-84.   DOI