Kanatlı Salmonella izolatlarının air thermal cycler amplifikasyonu ve kapiller jel elektroforezi ile hızlı identifikasyonu

The possible use of a genus-specific polymerase chain reaction (PCR) for the identification of Salmonella colonies was tested on 14 reference strains and 38 clinical Salmonella strains isolated from chickens by the standard method of the National Poultry Improvement Plan and Auxiliary Provisions (NPIP), USDA, U.S.A. All the PCRs using primer pairs InvA1 and InvA2 (5) gave 281 bp amplification product specific for the Salmonella genus and produced results identical (100%) to those obtained with biochemical and serological methods. Thus, the genus-specific capillary PCR for the identification of a colony of Salmonellae from a selective agar plate was performed within approximately 2 h and the total time required for definitive diagnosis of infection was 2 days using primary enrichment (PE) and 7 days using delayed secondary enrichment (DSE).

Rabid identification of avian Salmonella isolates by air thermal cycler amplification and capillary gel electrophoresis

The possible use of a genus-specific polymerase chain reaction (PCR) for the identification of Salmonella colonies was tested on 14 reference strains and 38 clinical Salmonella strains isolated from chickens by the standard method of the National Poultry Improvement Plan and Auxiliary Provisions (NPIP), USDA, U.S.A. All the PCRs using primer pairs InvA1 and InvA2 (5) gave 281 bp amplification product specific for the Salmonella genus and produced results identical (100%) to those obtained with biochemical and serological methods. Thus, the genus-specific capillary PCR for the identification of a colony of Salmonellae from a selective agar plate was performed within approximately 2 h and the total time required for definitive diagnosis of infection was 2 days using primary enrichment (PE) and 7 days using delayed secondary enrichment (DSE).

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  • 1.repka, M .J., Archer, J. R., Altekruse, S. F., Proctor, M. E, andDavis, J. P.: An increase in sporadic and outbreak-associatedSalmonella enteritidisinfections in Wisconsin: The Role of eggs., J.Infect. Dis., 1999,180: 1214-1219.
  • 2.Mishu, B., Koehler J., Lee, L. A., Rodrigue, D., Brenner. F. H.,Blake, P., and Tauxe, R. V.: Outbreaks of Salmonella enteritidisinfections in the United States, 1985-1991, J. Infect. Dis.,1994,169: 547-552.
  • 3.United States Department of Agriculture, National PoultryImprovement Plan and Auxiliary Provisions. U.S. Department ofAgriculture, Animal and Plant Health Inspection Service,Hyattsville, MD., 1994.
  • 4.Stefanovicova, A., Rehakova, H., Skarkova, A., Rijpens, N. andKuchta, T.: Confirmation of presumptive Salmonellacolonies bypolymerase chain reaction. J. Food Protect., 1998, 61: 1381-1383.
  • 5.Rahn, K., De Grandis, S. A., Clarke, R. C., McEwen, S. A., Galan,J. E., Ginocchio, C., Curtis III, R., and Gyles, C. L.: Amplification ofan invA gene sequence of Salmonella typhimuriumby polymerasechain reaction as a specific method of detection of Salmonella.Mol. Cell. Probes., 1992, 6: 271-279.
  • 6.Çarl›, K. T., Unal, C. B., Caner V., Eyigör A.: Detection ofSalmonellae in chicken feces by a combination of tetrathionatebroth enrichment, capillary PCR and capillary gel electrophoresis.J. Clin. Microbiol., (submitted for publication).
  • 7.Wittwer, C. T., Fillmore, G. C., and Hillyard, D. R.: Automatedpolymerase chain reaction in capillary tubes with hot air. NucleicAcid Res., 1989, 17: 4353-4357
Turkish Journal of Veterinary and Animal Sciences-Cover
  • ISSN: 1300-0128
  • Yayın Aralığı: Yılda 6 Sayı
  • Yayıncı: TÜBİTAK
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