Designing a rapid and accurate method for transportation and culture of the Campylobacter jejuni and Campylobacter coli-fastidious bacteria in the children with bacterial gastrointestinal symptoms

authors:

avatar Saeed Shams , avatar Bita Bakhshi , * , avatar Bahram Nikmanesh

Koomesh: Vol.18, issue 1; 71-78
published online: September 28, 2016
article type: issue_documents_importer
received: December 03, 2015
accepted: April 03, 2016

how to cite: Shams S, Bakhshi B, Nikmanesh B. Designing a rapid and accurate method for transportation and culture of the Campylobacter jejuni and Campylobacter coli-fastidious bacteria in the children with bacterial gastrointestinal symptoms. koomesh. 2016;18(1):e151147. 

Abstract

Introduction: Campylobacter spp. is the major cause of bacterial gastroenteritis, called campylobacteriosis, in the worldwide. Post-infectious complications of this infection are reactive arthritis and Guillain–Barré syndrome. Despite the importance of this infection, the isolation of fastidiousbacteria cannot be performed in most clinical laboratories. The aim this study was to design an alternative transport medium with mCCDA and evaluation the bacteria survival time into this medium, optimization of culture conditions of bacteria and then performance of direct duplex-PCR on colonies and stool samples. Finally, the results of the PCR and culture were compared. Materials and Methods: Fifty eight children suspected to campylobacteriosis were enrolled in this study. Fecal specimens were inoculated in depth inside the altered transport medium with mCCDA and then sent to laboratory. The specimens from transport media were cultured on two media of mCCDA;brucella agar daily and up to 7 days. Each plate was evaluated for bacterial growth up to 72 hours. The duplex-PCR on colonies and stool were carried out directly. Results: Total of isolated bacteria in this study was 9 cases (16%). The colonies were visible on the media after 48 to 72 h. The duplex-PCR assay on colonies detected 8 isolates of C. jejuni and 1 isolate C. coli. The results of the direct duplex-PCR on fecal specimens and cultures were the same. Conclusion: The results indicate that the presented method in this study with sensitivity equal to the PCR is useful for isolation of Campylobacter spp. It seems that using the changed transport medium and optimization of bacterial culture conditions will be isolated these fastidiousbacteria better than basic transport media.

References

  • 1.

    Eberle KN, Kiess AS. Phenotypic and genotypic methods for typing Campylobacter jejuni and Campylobacter coli in poultry. Poult Sci 2012; 91: 255-264.

  • 2.

    Cody AJ, McCarthy ND, Jansen van Rensburg M, Isinkaye T, Bentley SD, Parkhill J, et al. Real-time genomic epidemiological evaluation of human Campylobacter isolates by use of whole-genome multilocus sequence typing. J Clin Microb 2013; 51: 2526-2534.

  • 3.

    Korczak BM, Zurfluh M, Emler S, Kuhn-Oertli J, Kuhnert P. Multiplex strategy for multilocus sequence typing, fla typing, and genetic determination of antimicrobial resistance of Campylobacter jejuni and Campylobacter coli isolates collected in Switzerland. J Clin Microb 2009; 47: 1996-2007.

  • 4.

    Galanis E. Campylobacter and bacterial gastroenteritis. CMAJ 2007; 177: 570-571.

  • 5.

    Udayakumar D, Sanaullah M. Campylobacter cholecystitis. Int J Med Sci 2009; 6: 374-375.

  • 6.

    Smith JL. Campylobacter jejuni infection during pregnancy: long-term consequences of associated bacteremia, Guillain-Barre syndrome, and reactive arthritist. J Food Prot 2002; 65: 696-708.

  • 7.

    Waage AS, Vardund T, Lund V, Kapperud G. Detection of small numbers of Campylobacter jejuni and Campylobacter coli cells in environmental water, sewage, and food samples by a seminested PCR assay. Appl Environ Microbiol 1999; 65: 1636-1643.

  • 8.

    Miljkovic-Selimovic B, Lavrnic D, Moric O, Ng LK, Price L, Suturkova L, et al. Enteritis caused by Campylobacter jejuni followed by acute motor axonal neuropathy: a case report. J Med Case Rep 2010; 4: 101.

  • 9.

    Colmegna I, Cuchacovich R, Espinoza LR. HLA-B27-associated reactive arthritis: pathogenetic and clinical considerations. Clin Microbiol Rev 2004; 17: 348-369.

  • 10.

    (CDC) CfDCaP. Preliminary FoodNet Data on the incidence of infection with pathogens transmitted commonly through food--10 States, 2008. MMWR Morb Mortal Wkly Rep 2009; 58: 333-337.

  • 11.

    Eiland LS, Jenkins LS. Optimal treatment of campylobacter dysentery. J Pediatr Pharmacol Ther 2008; 13: 170-174.

  • 12.

    Lund M, Nordentoft S, Pedersen K, Madsen M. Detection of campylobacter spp. in chicken fecal samples by real-time PCR. J Clin Microb 2004; 42: 5125-5132.

  • 13.

    Poppert S, Haas M, Yildiz T, Alter T, Bartel E, Fricke U, Essig A. Identification of thermotolerant campylobacter species by fluorescence in situ hybridization. J Clin Microb 2008; 46: 2133-2136.

  • 14.

    Nachamkin I, Barbagallo S. Culture confirmation of Campylobacter spp. by latex agglutination. J Clin Microb 1990; 28: 817-818.

  • 15.

    Isenberg H. Clinical microbiology procedures handbook: American Society for Microbiology, Washington, DC; 2007.

  • 16.

    Shams S, Bakhshi B, Tohidi Moghadam T. In silico analysis of cadF gene and development of a duplex PCR for species-specific identification of Campylobacter jejuni and Campylobacter coli. Jundishapur J Microbiol 2016; 9: e29645.

  • 17.

    Yang JL, Wang MS, Cheng AC, Pan KC, Li CF, Deng SX. A simple and rapid method for extracting bacterial DNA from intestinal microflora for ERIC-PCR detection. World J Gastroenterol 2008; 14: 2872-2876.

  • 18.

    Singh H, Rathore RS, Singh S, Cheema PS. Comparative analysis of cultural isolation and PCR based assay for detection of campylobacter jejuni in food and faecal samples. Braz J Microbiol 2011; 42: 181-186.

  • 19.

    Masdor NA, Altintas Z, Tothill IE. Sensitive detection of Campylobacter jejuni using nanoparticles enhanced QCM sensor. Biosens Bioelectron 2016; 78: 328-336.

  • 20.

    Oh E, McMullen L, Jeon B. Impact of oxidative stress defense on bacterial survival and morphological change in Campylobacter jejuni under aerobic conditions. Front Microbiol 2015; 6: 295.

  • 21.

    Bolton FJ, Coates D, Hutchinson DN. The ability of campylobacter media supplements to neutralize photochemically induced toxicity and hydrogen peroxide. J Appl Bacteriol 1984; 56: 151-157.

  • 22.

    Huang H, Brooks BW, Lowman R, Carrillo CD. Campylobacter species in animal, food, and environmental sources, and relevant testing programs in Canada. Can J Microbiol 2015; 61: 701-721.

  • 23.

    Koneman EW. Koneman's Color Atlas and Textbook of Diagnostic Microbiology. 6th ed: Lippincott Williams Wilkins, Philadelphia, PA.; 2006.

  • 24.

    Salim SM, Mandal J, Parija SC. Isolation of Campylobacter from human stool samples. Indian J Med Microbiol 2014; 32: 35-38.

  • 25.

    Dallal MM, Khorramizadeh MR, MoezArdalan K. Occurrence of enteropathogenic bacteria in children under 5 years with diarrhoea in south Tehran. East Mediterr Health J 2006; 12: 792-797.

  • 26.

    Hassanzadeh P, Motamedifar M. Occurrence of Campylobacter jejuni in Shiraz, Southwest Iran. Med Princ Pract 2007; 16: 59-62.

  • 27.

    Ghorbanalizadgan M, Bakhshi B, Kazemnejad Lili A, Najar-Peerayeh S, Nikmanesh B. A molecular survey of Campylobacter jejuni and Campylobacter coli virulence and diversity. Iran Biomed J 2014; 18: 158-164.

  • 28.

    Platts-Mills JA, Liu J, Gratz J, Mduma E, Amour C, Swai N, et al. Detection of Campylobacter in stool and determination of significance by culture, enzyme immunoassay, and PCR in developing countries. J Clin Microb 2014; 52: 1074-1080.

  • 29.

    Al Amri A, Senok AC, Ismaeel AY, Al-Mahmeed AE, Botta GA. Multiplex PCR for direct identification of Campylobacter spp. in human and chicken stools. J Med Microbiol 2007; 56: 1350-1355##.