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The detection of hipO gene by real-time PCR in thermophilic Campylobacter spp. with very weak and negative reaction of hippurate hydrolysis

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Abstract

A total of 190 Campylobacter spp. isolates, of which 34 gave the result of very weak activity, and 156 gave the negative activity in the test for hippurate hydrolysis were characterized. The genomic DNA was isolated from a fresh culture of each isolate and the real-time PCR, targeting the hipO gene, was used to confirm the species distribution of Campylobacter isolates. The hipO gene was detected in 17 isolates (11%) within the total of 156 negative isolates for hippurate hydrolysis. Out of 34 isolates with very weak activity, 19 isolates (56%) were also found to be positive for hipO gene and characterized as C. jejuni. The real-time PCR assay used in this study could be employed for more accurate diagnosis of Campylobacter infections at species level after the biochemical characterization based on hippuricase activity of the isolates. This could also provide important data for the epidemiology of infections associated with these zoonotic pathogens.

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References

  • Blaser MJ, Taylor DN, Feldman RA (1983) Epidemiology of Campylobacter jejuni infections. Epidemiol Rev 5:157–176

    PubMed  CAS  Google Scholar 

  • Butzler JP (2004) Campylobacter from obscurity to celebrity. Clin Microbiol Infect 10:868–876. doi:10.1111/j.1469-0691.2004.00983.x

    Article  PubMed  Google Scholar 

  • Denis M, Soumet C, Rivaol K, Ermel G, Blivet D, Salvat G et al (1999) Development of a m-PCR assay for simultaneous identification of Campylobacter jejuni and C. coli. Lett Appl Microbiol 29:406–410. doi:10.1046/j.1472-765X.1999.00658.x

    Article  PubMed  CAS  Google Scholar 

  • El-Shibiny A, Connerton PL, Connerton IF (2005) Enumeration and diversity of campylobacters and bacteriophages isolated during the rearing cycles of free-range and organic chickens. Appl Environ Microbiol 71:1259–1266. doi:10.1128/AEM.71.3.1259-1266.2005

    Article  PubMed  CAS  Google Scholar 

  • Friedman CR, Hoekstra RM, Samuel M et al (2004) Risk factors for sporadic Campylobacter infection in the United States: a case–control study in FoodNet sites. Clin Infect Dis 38(Suppl 3):285–296. doi:10.1086/381598

    Article  Google Scholar 

  • Gorkiewicz G, Feierl G, Schober C, Dieber F, Kofer J, Zechner R et al (2003) Species-specific identification of campylobacters by partial 16S rRNA gene squencing. J Clin Microbiol 41:2537–2546. doi:10.1128/JCM.41.6.2537-2546.2003

    Article  PubMed  CAS  Google Scholar 

  • Hani EK, Chan VL (1995) Expression and characterization of Campylobacter jejuni benzoglycine aminohydrolase (hippuricase) gene in Escherichia coli. J Bacteriol 177:2396–2402

    PubMed  CAS  Google Scholar 

  • Hannu T, Mattila L, Rautelin H, Pelkonen P, Lahdenne P, Siitonen A et al (2002) Campylobacter-triggered reactive arthritis: a population-based study. Rheumatology 41:312–318. doi:10.1093/rheumatology/41.3.312

    Article  PubMed  CAS  Google Scholar 

  • Hughes RA, Rees JH (1997) Clinical and epidemiologic features of Guillain-Barre syndrome. J Infect Dis 176:92–98. doi:10.1086/513793

    Article  Google Scholar 

  • Hunt JM (1992) Campylobacter. In: Food and drug administration bacteriological analytical manual, 7th edn. Association of Official Analytical Chemists International, Arlington, VA, pp 77–94

  • Hwang M, Ederer GM (1975) Rapid hippurate hydrolysis method for the presumptive identification of group B streptococci. J Clin Microbiol 1:114–115

    PubMed  CAS  Google Scholar 

  • Kotsis IH, Adám M (1987) Paper-disc method for campylobacter hippurate-hydrolysis test. A note. Acta Microbiol Hung 34:173–177

    PubMed  CAS  Google Scholar 

  • Linton D, Lawson AJ, Owen RJ, Stanley J (1997) PCR detection, identification to species level, and fingerprinting of Campylobacter jejuni and Campylobacter coli direct from diarrheic samples. J Clin Microbiol 35:2568–2572

    PubMed  CAS  Google Scholar 

  • Mead PS, Slutsher L, Dietz V, McCaig LF, Bresse JS, Shapiro C et al (1999) Food-related illness and death in the United States. Emerg Infect Dis 5:840–842

    Article  Google Scholar 

  • Moore JE, Murphy PG (2000) Hippurate hydrolysis and speciation of thermophilic Campylobacter spp. Br J Biomed Sci 57:180–181

    PubMed  CAS  Google Scholar 

  • Morris GK, el Sherbeeny MR, Patton CM, Kodaka H, Lombard GL, Edmans P et al (1985) Comparison of four hippurate hydrolysis methods for identification of thermophilic Campylobacter spp. J Clin Microbiol 22:714–718

    PubMed  CAS  Google Scholar 

  • Nicholson MA, Patton CM (1995) Evaluation of disk method for hippurate hydrolysis by Campylobacter species. J Clin Microbiol 33:1341–1343

    PubMed  CAS  Google Scholar 

  • Nielsen EM, Engberg J, Madsen M (1997) Distribution of serotypes of Campylobacter jejuni and C. coli from Danish patients, poultry, cattle and swine. FEMS Immunol Med Microbiol 19:47–56

    PubMed  CAS  Google Scholar 

  • On SLW (1996) Identification methods for campylobacters, helicobacters, and related organisms. Clin Microbiol Rev 9:405–422

    PubMed  CAS  Google Scholar 

  • On SLW, Holmes B (1991) Effect of inoculum size on the phenotypic characterization of Campylobacter species. J Clin Microbiol 29:923–926

    PubMed  CAS  Google Scholar 

  • On SLW, Holmes B (1992) Assessment of enzyme detection tests useful in identification of campylobacteria. J Clin Microbiol 30:746–749

    PubMed  CAS  Google Scholar 

  • Rautelin H, Jusufovic J, Hänninen M-L (1999) Identification of hippurate-negative thermophilic campylobacters. Diagn Microbiol Infect 35:9–12. doi:10.1016/S0732-8893(99)00057-7

    Article  CAS  Google Scholar 

  • Rönner A-C, Engvall EO, Andersson L, Kaijser B (2004) Species identification by genotyping and determination of antibiotic resistance in Campylobacter jejuni and Campylobacter coli from humans and chickens in Sweden. Int J Food Microbiol 96:173–179. doi:10.1016/j.ijfoodmicro.2004.03.017

    Article  PubMed  CAS  Google Scholar 

  • Rudi K, Høidal HK, Katla T, Johansen BK, Nordal J, Jakobsen KS (2004) Direct real-time PCR quantification of Campylobacter jejuni in chicken fecal and cecal samples by integrated cell concentration and DNA purification. Appl Environ Microbiol 70:790–797. doi:10.1128/AEM.70.2.790-797.2004

    Article  PubMed  CAS  Google Scholar 

  • Schlundt J, Toyofuku H, Jansen J, Herbst SA (2004) Emerging food-borne zoonoses. Rev Sci Tech 23:513–533

    PubMed  CAS  Google Scholar 

  • Skirrow MB, Benjamin J (1980) Differentiation of enteropathogenic campylobacter. J Clin Pathol 33:1122. doi:10.1136/jcp.33.11.1122

    Article  PubMed  CAS  Google Scholar 

  • Steinhauserova I, Češkova J, Fojtikova K, Obrovska I (2001) Identification of thermophilic Campylobacter spp. by phenotypic and molecular methods. J Appl Microbiol 90:470–475. doi:10.1046/j.1365-2672.2001.01267.x

    Article  PubMed  CAS  Google Scholar 

  • Taylor LH, Latham SM, Woolhouse ME (2001) Risk factors for human disease emergence. Philos Trans R Soc B 29:983–989

    Google Scholar 

  • Totten PA, Patton CM, Tenover FC, Barrett TJ, Stamm WE, Steigerwalt AG et al (1987) Prevalance and characterization of hippurate-negative Campylobacter jejuni in King County, Washington. J Clin Microbiol 25:1747–1752

    PubMed  CAS  Google Scholar 

  • Wainø M, Bang DD, Lund M, Nordentoft S, Andersen JS, Pedersen K et al (2003) Identification of campylobacteria isolated from Danish broilers by phenotypic tests and species-specific PCR assays. J Appl Microbiol 95:649–655. doi:10.1046/j.1365-2672.2003.01996.x

    Article  PubMed  CAS  Google Scholar 

  • Yang C, Jiang Y, Huang K, Zhu C, Yin Y (2003) Application of real-time PCR for quantitative detection of Campylobacter jejuni in poultry, milk and environmental water. FEMS Immunol Med Microbiol 38:265–271. doi:10.1016/S0928-8244(03)00168-8

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

The authors wish to acknowledge to Can Akar for technical assistance. This study was supported by grant 104T242 from The Scientific and Technological Research Council of Turkey, TUBITAK.

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Correspondence to Vildan Caner.

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Caner, V., Cokal, Y., Cetin, C. et al. The detection of hipO gene by real-time PCR in thermophilic Campylobacter spp. with very weak and negative reaction of hippurate hydrolysis. Antonie van Leeuwenhoek 94, 527–532 (2008). https://doi.org/10.1007/s10482-008-9269-4

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