Advertisement

Archives of Microbiology

, Volume 200, Issue 7, pp 1009–1016 | Cite as

Rapid molecular identification and differentiation of common Salmonella serovars isolated from poultry, domestic animals and foodstuff using multiplex PCR assay

  • A. B. Alzwghaibi
  • R. Yahyaraeyat
  • B. Nayeri Fasaei
  • A. Ghalyanchi Langeroudi
  • T. Zahraei Salehi
Original Paper

Abstract

Salmonella is widely distributed throughout the world and can be found in poultry industry, animal breeding centers, food and feedstuffs of all geographical regions. This study was conducted to determine and identify Salmonella serovars isolated from poultry, calves and foodstuffs (poultry and animals products such as egg and meat). A total of one hundred isolates of Salmonella serovars including Salmonella Typhimurium, Salmonella Enteritidis, Salmonella Infantis, Salmonella Gallinarum and Salmonella Pullorum consecutively were subjected to the conventional culture, biochemical and serological assays. The utility of molecular multiplex PCR was investigated to identify and differentiate among five Salmonella serovars which were identified according to the presence of rfbJ, fljB, invA, and fliC genes in S. Typhimurium, sefA, invA and spv genes in Salmonella Enteritidis, fljB, fliC and invA genes in Salmonella Infantis, hut and slgC genes in both Salmonella Gallinarum and Salmonella Pullorum and speC gene specifically in Salmonella Gallinarum. Biochemical assays and serotyping are complicated to directly differentiate between Salmonella Gallinarum and Salmonella Pullorum because of their antigenic similarity. According to the results, Multiplex PCR can be considered as simple, rapid, accurate and useful test to identify and differentiate among Salmonella serovars.

Keywords

Salmonella serovars Differentiation Genotyping Salmonella genes Multiplex PCR 

Notes

Acknowledgements

This work was supported by the University of Tehran, Faculty of Veterinary Medicine, Grant No. 7504002. The authors would like to thanks Mr. Iradj Ashrafi Tamai for the cooperation in laboratory process.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Informed consent

This article does not contain any studies with human participants or animals performed by any of the authors.

References

  1. Akiba M, Kusumoto M, Iwata T (2011) Rapid identification of Salmonella enterica serovars, typhimurium, choleraesuis, infantis, hadar, enteritidis, dublin and gallinarum, by multiplex PCR. J Microbiol Meth 85:9–15CrossRefGoogle Scholar
  2. Badouei MA, Ghalejooghi BM, Madadgar O (2012) Study on Salmonella contamination of traditionally produced edible poultry eggs. Comp Clin Pathol 21:1093–1097CrossRefGoogle Scholar
  3. Bangtrakulnonth A et al. (2004) Salmonella serovars from humans and other sources in Thailand, 1993–2002. Emerg Infect Dis 10:131CrossRefPubMedPubMedCentralGoogle Scholar
  4. Barrow P, Neto OF (2011) Pullorum disease and fowl typhoid—new thoughts on old diseases: a review. Avian Pathol 40:1–13CrossRefPubMedGoogle Scholar
  5. Barrow P, Jones M, Smith A, Wigley P (2012) The long view: Salmonella—the last forty years. Avian Pathol 41:413–420CrossRefPubMedGoogle Scholar
  6. Batista DFA, de Freitas Neto OC, Lopes PD, de Almeida AM, Barrow PA, Berchieri A Jr (2013) Polymerase chain reaction assay based on ratA gene allows differentiation between Salmonella enterica subsp. enterica serovar Gallinarum biovars Gallinarum and Pullorum. J Vet Diag Invest 25:259–262CrossRefGoogle Scholar
  7. Borges KA, Furian TQ, Borsoi A, Moraes HL, Salle CT, Nascimento VP (2013) Detection of virulence-associated genes in Salmonella Enteritidis isolates from chicken in South of Brazil. Pesquisa Veterinaria Brasileira 33:1416–1422CrossRefGoogle Scholar
  8. Carter G, Wise D, Carter G (2004) Essentials of veterinary bacteriology and mycology, 6th edn. Iowa State University Press, AmesGoogle Scholar
  9. Dilmaghani M, Ahmadi M, Zahraei-Salehi T, Talebi A (2011) Detection of Salmonella enterica serovar Typhimurium from avians using multiplex-PCR. Vet Res Forum 2:157–165Google Scholar
  10. Dinarvand R, Sepehri N, Manoochehri S, Rouhani H, Atyabi F (2011) Polylactide-co-glycolide nanoparticles for controlled delivery of anticancer agents. Int J Nanomed 6:877CrossRefGoogle Scholar
  11. Dousandeh S, Asaadi Tehrani G, Amini B, Maziri P (2016) Simultaneous detection of invA, STM4497, and fliC183 genes in Salmonella typhimurium by multiplex PCR method in poultry meat samples in Zanjan, Iran. Infect Epidemiol Med 2:20–23CrossRefGoogle Scholar
  12. Emaddi Chashni S, Hassanzadeh M, Bozorgmehri Fard M, Mirzaie S (2009) Characterization of the Salmonella isolates from backyard chickens in north of Iran, by serotyping, multiplex PCR and antibiotic resistance analysis. Archives of Razi Institute, KarajGoogle Scholar
  13. Farkas J, Doyle M, Beuchat L, Montville T (2001) Food microbiology: fundamentals and frontiers. ASM Publication, Washington, DCGoogle Scholar
  14. Ghoddusi A, Fasaei BN, Karimi V, Tamai IA, Moulana Z, Salehi TZ (2015) Molecular identification of Salmonella Infantis isolated from backyard chickens and detection of their resistance genesby PCR. Iran J Vet Res 16:293PubMedPubMedCentralGoogle Scholar
  15. Hashemi A, Baghbani-Arani F (2015) The effective differentiation of Salmonella isolates using four PCR-based typing methods. J Appl Microbiol 118:1530–1540CrossRefPubMedGoogle Scholar
  16. Herikstad H, Motarjemi Y, Tauxe R (2002) Salmonella surveillance: a global survey of public health serotyping. Epidemiol Infect 129:1–8PubMedPubMedCentralGoogle Scholar
  17. Herrera-León S, Ramiro R, Arroyo M, Díez R, Usera MA, Echeita MA (2007) Blind comparison of traditional serotyping with three multiplex PCRs for the identification of Salmonella serotypes. Res Microbiol 158:122–127CrossRefPubMedGoogle Scholar
  18. Ibrahim H, Abd El-moaty DAM, Ahmed HA, El-Enbaawy MI (2016) Phenotypic and genotypic characterization of locally isolated Salmonella strains used in preparation of Salmonella antigens in Egypt. Vet World 9:1435–1439CrossRefPubMedPubMedCentralGoogle Scholar
  19. Issenhuth-Jeanjean S et al (2014) Supplement 2008–2010 (no. 48) to the White–Kauffmann–Le Minor scheme. Res Microbiol 165:526–530CrossRefPubMedGoogle Scholar
  20. Jalali M, Abedi D, Pourbakhsh SA, Ghoukasin K (2008) Prevalence of salmonella spp. in raw and cooked foods in Isfahan-Iran. J Food Safe 28:442–452CrossRefGoogle Scholar
  21. Kang MSKY., Jung BY, Kim A, Lee KM, An BK, Song EA, Kwon JH, Chung GS (2011) Differential dentification of Salmonella enterica subsp. enterica serovar Gallinarum biovars Gallinarum and Pullorum based on polymorphic regions of glgC and speC genes. Vet Microbiol 147:181–185CrossRefPubMedGoogle Scholar
  22. Kardos G, Farkas T, Antal M, Nogrady N, Kiss I (2007) Novel PCR assay for identification of Salmonella enterica serovar Infantis. Lett Appl Microbiol 45:421–425CrossRefPubMedGoogle Scholar
  23. Kwon H-J, Park K-Y, Yoo H-S, Park J-Y, Park YH, Kim S-J (2000) Differentiation of Salmonella enterica serotype Gallinarum biotype Pullorum from biotype Gallinarum by analysis of phase 1 flagellin C gene (fliC). J Microbiol Meth 40:33–38CrossRefGoogle Scholar
  24. Li X, Payne J, Santos F, Levine J, Anderson K, Sheldon B (2007) Salmonella populations and prevalence in layer feces from commercial high-rise houses and characterization of the Salmonella isolates by serotyping, antibiotic resistance analysis, and pulsed field gel electrophoresis. Poultry Sci 86:591–597CrossRefGoogle Scholar
  25. Li R, Wang Y, Shen J, Wu C (2014) Development of a novel hexa-plex PCR method for identification and serotyping of Salmonella species. Foodborne Pathogens dis 11:75–77CrossRefGoogle Scholar
  26. Lim Y-H et al (2003) Multiplex polymerase chain reaction assay for selective detection of Salmonella enterica serovar Typhimurium. Japan J Infect Dis 56:151–155Google Scholar
  27. Lin YK, YK.Hu, SC Hua, Z. Meng, G Xu, XC Juan, CS Rong, W Ming, CJ Xin (2014) Development-of a multiplex PCR for rapid identification of Salmonella Enteritidis, Salmonella Typhimurium, Salmonella Pullorum and Salmonella Gallinarum. Acta Veterinaria et Zootechnica Sinica 45:268–273Google Scholar
  28. McQuiston J, Parrenas R, Ortiz-Rivera M, Gheesling L, Brenner F, Fields P (2004) Sequencing and comparative analysis of flagellin genes fliC, fljB, and flpA from Salmonella. J Clin Microbiol 42:1923–1932CrossRefPubMedPubMedCentralGoogle Scholar
  29. Miller T, Prager R, Rabsch W, Fehlhaber K, Voss M (2010) Epidemiological relationship between Salmonella Infantis isolates of human and broiler origin. Lohmann Inf 45:27-31Google Scholar
  30. Moosavy M-H, Esmaeili S, Amiri FB, Mostafavi E, Salehi TZ (2015) Detection of Salmonella spp in commercial eggs in Iran. Iran J Microbiol 7:50PubMedPubMedCentralGoogle Scholar
  31. Nagappa K, Tamuly S, Saxena M, Singh S (2007) Isolation of Salmonella Typhimurium from poultry eggs and meat of Tarai region of Uttaranchal. Indian J Biotechnol 6:407-409Google Scholar
  32. Oliveira S, Santos L, Schuch D, Silva A, Salle C, Canal C (2002) Detection and identification of salmonellas from poultry-related samples by PCR. Vet Microbiol 87:25–35CrossRefPubMedGoogle Scholar
  33. Popoff MY, Bockemühl J, Gheesling LL (2004) Supplement 2002 (no. 46) to the Kauffmann–White scheme. Res Microbiol 155:568–570CrossRefPubMedGoogle Scholar
  34. Rahn K et al (1992) Amplification of an invA gene sequence of Salmonella typhimurium by polymerase chain reaction as a specific method of detection of Salmonella. Mol Cell Probes 6:271–279CrossRefPubMedGoogle Scholar
  35. Raut S (2014) Pathology of salmonella enterica challenge infection. In: Broilers and its amelioration by bacillus subtilis supplementation. MAFSUGoogle Scholar
  36. Ribeiro SAM, Paiva JBd, Zotesso F, Lemos MVF, Berchieri Júnior  (2009) Molecular differentiation between Salmonella enterica subsp enterica serovar Pullorum and Salmonella enterica subsp enterica serovar Gallinarum. Brazil J Microbiol 40:184–188CrossRefGoogle Scholar
  37. Salehi TZ, Mahzounieh M, Saeedzadeh A (2005) Detection of invA gene in isolated Salmonella from broilers by PCR method. Int J Poult Sci 4:557–559CrossRefGoogle Scholar
  38. Shivaprasad H (2000) Fowl typhoid and pullorum disease. Revue Sci et Tech (Int Off Epizootics) 19:405–424CrossRefGoogle Scholar
  39. Soumet C et al (1999) Evaluation of a multiplex PCR assay for simultaneous identification of Salmonella sp., Salmonella Enteritidis and Salmonella Typhimurium from environmental swabs of poultry houses. Lett Appl Microbiol 28:113–117CrossRefPubMedGoogle Scholar
  40. Swayne DE (1998) Laboratory manual for the isolation and identification of avian pathogens. American Association of Avian Pathologists. University of Pennsylvania, PhiladelphiaGoogle Scholar
  41. Woodward MJ, Kirwan S (1996) Detection of Salmonella enteritidis in eggs by the polymerase chain reaction. Vet Rec 138:411–413CrossRefPubMedGoogle Scholar
  42. Zahraei Salehi T, Tadjbakhsh H, Atashparvar N, Nadalian M, Mahzounieh M (2007) Detection and identification of Salmonella Typhimurium in bovine diarrhoeic fecal samples by immunomagnetic separation and multiplex PCR assay. Zoonoses Public Health 54:231–236CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • A. B. Alzwghaibi
    • 1
    • 2
  • R. Yahyaraeyat
    • 1
  • B. Nayeri Fasaei
    • 1
  • A. Ghalyanchi Langeroudi
    • 1
  • T. Zahraei Salehi
    • 1
  1. 1.Department of Microbiology and Immunology, Faculty of Veterinary MedicineUniversity of TehranTehranIran
  2. 2.Department of Animal Source, Faculty of AgricultureUniversity of Al-Qasim GreenAl-QasimIraq

Personalised recommendations