Performance of five molecular methods for monitoring Arcobacter spp

BMC Microbiology

María José Figueras¹  Arturo Levican¹ 

[1] Unitat de Microbiologia, Departament de Ciències Mediques Bàsiques, Facultat de Medicina i Ciències de la Salut. IISPV, Universitat Rovira i Virgili, Reus, Spain

关键词: gyrA;    23S rRNA;    m-PCR;    16S rRNA-RFLP;    Molecular methods;    Comparison;    Identification;    Arcobacter;   

Others  :  1143000
DOI  :  10.1186/1471-2180-13-220

 received in 2013-02-07, accepted in 2013-09-27,  发布年份 2013

【 摘 要 】

Background

Bacteria belonging to the Arcobacter genus are emerging enteropathogens and potential zoonotic agents. Their taxonomy has evolved very rapidly, and there are presently 18 recorded species. The prevalence of species belonging to Arcobacter is underestimated because of the limitations of currently available methods for species identification.

The aim of this study was to compare the performance of five PCR based methods that target regions of 16S rRNA, 23S rRNA or gyrA genes to identify Arcobacter species, and to review previous results reported in the literature using these methods.

Results

The five tested methods were found not to be reliable. They misidentified between 16.8% and 67.4% of the studied strains; this was dependent upon the target regions of the tested genes. The worst results obtained were for the identification of Arcobacter cryaerophilus and Arcobacter butzleri when the 23S rRNA gene was used as the target. These species were confused with many non-targeted species.

Conclusion

Our results suggest that the known diversity of Arcobacter spp. in different environments could be expanded if reliable identification methods are applied in future studies.

【 授权许可】

   
2013 Levican and Figueras; licensee BioMed Central Ltd.

【 参考文献 】

• [1]Collado L, Figueras MJ: Taxonomy, epidemiology and clinical relevance of the genus Arcobacter. Clin Microbiol Rev 2011, 24:174-192.
• [2]Collado L, Inza I, Guarro J, Figueras MJ: Presence of Arcobacter spp. in environmental waters correlates with high levels of fecal pollution. Environ Microbiol 2008, 10:1635-1640.
• [3]Collado L, Kasimir G, Perez U, Bosch A, Pinto R, Saucedo G, Huguet JM, Figueras MJ: Occurrence and diversity of Arcobacter spp. along the Llobregat river catchment, at sewage effluents and in a drinking water treatment plant. Water Res 2010, 44:3696-3702.
• [4]Vandamme P, Falsen E, Rossau R, Hoste B, Segers P, Tytgat R, De Ley J: Revision of Campylobacter, Helicobacter, and Wolinella taxonomy: emendation of generic descriptions and proposal of Arcobacter gen. nov. Int J Syst Bacteriol 1991, 41:88-103.
• [5]Figueras MJ, Levican A, Collado L, Inza MI, Yustes C: Arcobacter ellisii sp. nov., isolated from mussels. Syst Appl Microbiol 2011, 34:414-418.
• [6]Levican A, Collado L, Aguilar C, Yustes C, Diéguez AL, Romalde JL, Figueras MJ: Arcobacter bivalviorum sp. nov. and Arcobacter venerupis sp. nov., new species isolated from shellfish. Syst Appl Microbiol 2012, 35:133-138.
• [7]Levican A, Collado L, Figueras MJ: Arcobacter cloacae sp. nov. and Arcobacter suis sp. nov., new species isolated from food and sewage. Syst Appl Microbiol 2013, 36:22-27.
• [8]Sasi Jyothsna TS, Rahul K, Ramaprasad EV, Sasikala C, Ramana CV: Arcobacter anaerophilus sp. nov., isolated from an estuarine sediment and emended description of the genus Arcobacter. Int J Syst Evol Microbioldoi:10.1099/ijs.0.054155-0. In press
• [9]Douidah L, De Zutter L, Vandamme P, Houf K: Identification of five human and mammal associated Arcobacter species by a novel multiplex-PCR assay. J Microbiol Methods 2010, 80:281-286.
• [10]Bastyns K, Cartuyvelsi D, Chapelle S, Vandamme P, Goosens H, De Watcher R: A variable 23S rDNA region is a useful discriminating target for genus-specific and species-specific PCR amplification in Arcobacter species. Syst Appl Microbiol 1995, 18:353-356.
• [11]Moreno Y, Botella S, Alonso JL, Ferrus MA, Hernandez M, Hernandez J: Specific detection of Arcobacter and Campylobacter strains in water and sewage by PCR and fluorescent in situ hybridization. Appl Env Microbiol 2004, 69:1181-1186.
• [12]Abdelbaqi K, Buissonniere A, Prouzet-Mauleon V, Gresser J, Wesley I, Mégraud F, Ménard A: Development of a real-time fluorescence resonance energy transfer PCR to detect Arcobacter species. J Clin Microbiol 2007, 45:3015-3021.
• [13]González A, Moreno Y, Gonzalez R, Hernández J, Ferrus MA: Development of a simple and rapid method based on polymerase chain reaction-based restriction fragment length polymorphism analysis to differentiate Helicobacter, Campylobacter, and Arcobacter species. Curr Microbiol 2006, 53:416-421.
• [14]Houf K, Tutenel A, De Zutter L, Van Hoof J, Vandamme P: Development of a multiplex PCR assay for the simultaneous detection and identification of Arcobacter butzleri, Arcobacter cryaerophilus and Arcobacter skirrowii. FEMS Microbiol Lett 2000, 193:89-94.
• [15]Kabeya H, Kobayashi Y, Maruyama S, Mikami T: Distribution of Arcobacter species among livestock in Japan. Vet Microbiol 2003, 93:153-158.
• [16]Pentimalli D, Pegels N, Garcia T, Martin R, González I: Specific PCR detection of Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter skirrowii, and Arcobacter cibarius in chicken meat. J Food Prot 2009, 72:1491-1495.
• [17]De Smet S, Vandamme P, De Zutter L, On S, Douidah L, Houf K: Arcobacter trophiarum sp. nov. isolated from fattening pigs. Int J Syst Evol Microbiol 2011, 63:356-361.
• [18]Figueras MJ, Collado L, Guarro J: A new 16S rDNA-RFLP method for the discrimination of the accepted species of Arcobacter. Diagn Microbiol Infect Dis 2008, 62:11-15.
• [19]Figueras MJ, Levican A, Collado L: Updated 16S rRNA-RFLP method for the identification of all currently characterized Arcobacter spp. BMC Microbiol 2012, 12:292. BioMed Central Full Text
• [20]Liberati A, Altman DG, Tetzlaff J, Mulrow C, Gøtzsche PC, Loannidis JPA, Clarke M, Devereaux PJ, Kleijnen J, Moher D: The PRISMA statement for reporting systematic reviews and meta-analyses of studies that evaluate health care interventions: explanation and elaboration. PLoS Med 2009, 6(7):e1000100.
• [21]Debruyne L, Houf K, Douidah L, De Smet S, Vandamme P: Reassessment of the taxonomy of Arcobacter cryaerophilus. Syst Appl Microbiol 2010, 33:7-14.
• [22]Atabay HI, Waino M, Madsen M: Detection and diversity of various 754 Arcobacter species in Danish poultry. Int J Food Microbiol 2006, 109:139-145.
• [23]Collado L, Guarro J, Figueras MJ: Prevalence of Arcobacter in meat and shellfish. J Food Prot 2009, 72:1102-1106.
• [24]Collado L, Cleenwerck I, Van Trappen S, De Vos P, Figueras MJ: Arcobacter mytili sp. nov., an indoxyl acetate-hydrolysis-negative bacterium isolated from mussels. Int J Syst Evol Microbiol 2009, 59:1391-1396.
• [25]Figueras MJ, Collado L, Levican A, Perez J, Solsona MJ, Yustes C: Arcobacter molluscorum sp. nov., new species isolated from shellfish. Syst Appl Microbiol 2011, 34:105-109.
• [26]Collado L, Levican A, Perez J, Figueras MJ: Arcobacter defluvii sp. nov., isolated from sewage. Int J Syst Evol Microbiol 2011, 61:1895-1901.
• [27]De Smet S, De Zutter L, Debruyne L, Vangroenweghe F, Vandamme P, Houf K: Arcobacter population dynamics in pigs on farrow-to-finish farms. Appl Environ Microbiol 2011, 77:1732-1738.
• [28]De Smet S, De Zutter L, Houf K: Small ruminants as carriers of the emerging foodborne pathogen Arcobacter on small and medium farms. Small Ruminants Res 2011, 97:124-129.