A Rapid Identification of Salmonella enterica in Chicken Skin Using an invA Molecular Marker

Journal of Cell and Molecular Research

مقاله 4، دوره 15، شماره 1 - شماره پیاپی 29، دی 2023، صفحه 34-39   

نوع مقاله: Research Articles

شناسه دیجیتال (DOI): 10.22067/jcmr.2023.84013.1076

نویسندگان

Najmeh Sodagar¹؛ Ahmad Reza Bahrami^{* 1، 2}؛ Maryam Moghaddam Matin^{1، 3}

¹Department of Biology, Faculty of science, Ferdowsi University of Mashhad, Mashhad, Iran

²Industrial Biotechnology Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran

³Novel Diagnostics and Therapeutics Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran

چکیده

Salmonella is a gram-negative bacillus that lives in the intestinal tract of human and animals and causes diarrhea. Salmonella could be found in undercooked products of poultry with no impact on the taste, smell, or appearance. Since poultry eggs and meat might be sources of Salmonella and pose a hazard to public health, it is important to accurately detect Salmonella infection. In this regard, the present study aimed to develop a rapid and sensitive method for the diagnosis of Salmonella spp. in samples from the poultry industry. To do so, the sensitivity of S. enterica serotype Enteritidis detection was assessed with ten-fold serial dilutions in peptone water to give suspensions containing 10⁰ to 10⁵ CFU/mL. For artificial inoculation, skin samples were sequentially inoculated with the serial dilutions, while a control sample was included to ensure that the skin was not naturally contaminated with Salmonella. 53 commercial chicken skin samples were obtained from different local shops. Then, DNA was extracted from all samples, and the quality of extracted DNAs was checked by spectrophotometry and confirmed by agarose gel electrophoresis. For PCR, a pair of oligonucleotide primers, INVA, was designed to amplify the invA gene. Results revealed a band of 796 bp in samples artificially contaminated with S. Enteritidis. Likewise, the 796 bp band was detected in 38 samples (71%) with deferent intensities, which presented different amounts of contamination. Accordingly, the present study provided a valuable method for the detection and control of Salmonella infection in the poultry industry, since results would be available in less time than with the conventional cultural method.

کلیدواژه‌ها

Salmonella enterica؛ invA gene؛ PCR assay؛ public health

مراجع

Aabo S., Andersen J. and Olsen J. (1995) Research note: detection of Salmonella in minced meat by the polymerase chain reaction method. Letters in Applied Microbiology 21:180-182.
Araj G. and Chugh T. D. (1987) Detection of Salmonella spp. in clinical specimens by capture enzyme-linked immunosorbent assay. Journal of clinical microbiology 25:2150-2153.
Aspinall S., Hindle M. and Hutchinson D. (1992) Improved isolation of salmonellae from faeces using a semisolid Rappaport-Vassiliadis medium. European journal of clinical microbiology and infectious diseases 11:936-939.
Bennett A., Greenwood D., Tennant C., Banks J. and Betts R. (1998) Rapid and definitive detection of Salmonella in foods by PCR. Letters in Applied Microbiology 26:437-441.
Braun S. D. and Methner U. (2011) Comparison of DNA isolation methods and detection of Salmonella spp. from animal faeces and dust using invA real-time PCR. Berl Munch Tierarztl Wochenschr 124:177-185.
Callaway T. R., Edrington T., Anderson R. C., Byrd J. A. and Nisbet D. J. (2008) Gastrointestinal microbial ecology and the safety of our food supply as related to Salmonella. Journal of Animal Science 86:E163-E172.
Chen S., Yee A., Griffiths M., Wu K., Wang C. N., Rahn K. and De Grandis S. (1997) A rapid, sensitive and automated method for detection of Salmonella species in foods using AG‐9600 AmpliSensor Analyzer. Journal of Applied Microbiology 83:314-321.
Chiu C.-H. and Ou J. T. (1996) Rapid identification of Salmonella serovars in feces by specific detection of virulence genes, invA and spvC, by an enrichment broth culture-multiplex PCR combination assay. Journal of clinical microbiology 34:2619-2622.
Cohen N., Martin L., Simpson R., Wallis D. and Neibergs H. (1996) Comparison of polymerase chain reaction and microbiological culture for detection of salmonellae in equine feces and environmental samples. American journal of veterinary research 57:780-786.
de Freitas C. G., Santana Â. P., da Silva P. H. C., Gonçalves V. S. P., Barros M. d. A. F., Torres F. A. G., Murata L. S. and Perecmanis S. (2010) PCR multiplex for detection of Salmonella Enteritidis, Typhi and Typhimurium and occurrence in poultry meat. International Journal of Food Microbiology 139:15-22.
Fratamico P. and Strobaugh T. (1998) Simultaneous detection of Salmonella spp and Escherichia coli O157: H7 by multiplex PCR. Journal of Industrial Microbiology and Biotechnology 21:92-98.
Galán J. E. and Curtiss 3rd R. (1991) Distribution of the invA,-B,-C, and-D genes of Salmonella typhimurium among other Salmonella serovars: invA mutants of Salmonella typhi are deficient for entry into mammalian cells. Infection and immunity 59:2901-2908.
Gallegos‐Robles M., Morales‐Loredo A., Álvarez‐Ojeda G., Osuna‐Garcia J., Martinez I., Morales‐Ramos L. and Fratamico P. (2009) PCR detection and microbiological isolation of Salmonella spp. from fresh beef and cantaloupes. Journal of Food Science 74:M37-M40.
Germini A., Masola A., Carnevali P. and Marchelli R.(2009) Simultaneous detection of Escherichia coliO175: H7, Salmonella spp., and Listeriamonocytogenes by multiplex PCR. Food Control20:733-738.
Gouws P. A., Visser M. and Brözel V. S. (1998) A polymerase chain reaction procedure for the detection of Salmonella spp. within 24 hours. Journal of food protection 61:1039-1042.
Guiney D. G., Fang F. C., Krause M., Libby S., Buchmeier N. A., Fierer J., Fang F. C., Krause M., Libby S. and Buchmeier N. A. (1995) Biology and clinical significance of virulence plasmids in Salmonella serovars. Clinical infectious diseases 21:S146-S151.
Gulig P. A., Caldwell A. L. and Chiodo V. A. (1992) Identification, genetic analysis and DNA sequence of a 7.8‐kb virulence region of the Salmonella typhimurium virulence plasmid. Molecular microbiology 6:1395-1411.
Gulig P. A., Danbara H., Guiney D. G., Lax A. J., Norel F. and Rhen M. (1993) Molecular analysis of spv virulence genes of the Salmonella virulence plasmids. Molecular microbiology 7:825-830.
Khnouf R., Jaradat M. A. K., Karasneh D., Al-Shami F., Sawaqed L. and Albiss B. A. (2020) Simulation and optimization of a single heater convective PCR chip and its controller for fast salmonella Enteritidis detection. IEEE Sensors Journal 20:13186-13195.
Kimura B., Kawasaki S., Fujii T., Kusunoki J., Itoh T.and Flood S. J. (1999) Evaluation of TaqMan PCR assay for detecting Salmonella in raw meat and shrimp. Journal of food protection 62:329-335.
Klerks M., Van Bruggen A., Zijlstra C. and Donnikov M. (2006) Comparison of methods of extracting Salmonella enterica serovar Enteritidis DNA from environmental substrates and quantification of organisms by using a general internal procedural control. Applied and Environmental Microbiology 72:3879-3886.
Lin J.-S. and Tsen H.-Y. (1999) Development and use of polymerase chain reaction for the specific detection of Salmonella Typhimurium in stool and food samples. Journal of food protection 62:1103-1110.
Luk J. M., Kongmuang U., Tsang R. and Lindberg A. A. (1997) An enzyme-linked immunosorbent assay to detect PCR products of the rfbS gene from serogroup D salmonellae: a rapid screening prototype. Journal of clinical microbiology 35:714-718.
Maciorowski K., Pillai S., Jones F. and Ricke S. (2005) Polymerase chain reaction detection of foodborne Salmonella spp. in animal feeds. Critical reviews in microbiology 31:45-53.
Malorny B., Hoorfar J., Bunge C. and Helmuth R. (2003) Multicenter validation of the analytical accuracy of Salmonella PCR: towards an international standard. Applied and Environmental Microbiology 69:290-296.
Nazari Moghadam M., Rahimi E., Shakerian A. and Momtaz H. (2023) Prevalence of Salmonella Typhimurium and Salmonella Enteritidis isolated from poultry meat: virulence and antimicrobial-resistant genes. BMC microbiology 23:1-8.
Rijpens N., Herman L., Vereecken F., Jannes G., De Smedt J. and De Zutter L. (1999) Rapid detection of stressed Salmonella spp. in dairy and egg products using immunomagnetic separation and PCR. International Journal of Food Microbiology 46:37-44.
Silva D. S. P., Canato T., Magnani M., Alves J., Hirooka E. Y. and de Oliveira T. C. R. M. (2011) Multiplex PCR for the simultaneous detection of Salmonella spp. and Salmonella Enteritidis in food. International journal of food science & technology 46:1502-1507.
Soumet C., Ermel G., Rose V., Rose N., Drouin P., Salvat G. and Colin P. (1999) Identification by a multiplex PCR‐based assay of Salmonella Typhimurium and Salmonella Enteritidis strains from environmental swabs of poultry houses. Letters in Applied Microbiology 29:1-6.
Stone G. G., Oberst R. D., Hays M. P., Mcvey S. and Chengappa M. (1994) Detection of Salmonella serovars from clinical samples by enrichment broth cultivation-PCR procedure. Journal of clinical microbiology 32:1742-1749.
Stone G. G., Oberst R. D., Hays M. P., McVey S., Galland J. C., Curtiss 3rd R., Kelly S. M. and Chengappa M. (1995) Detection of Salmonella typhimurium from rectal swabs of experimentally infected beagles by short cultivation and PCR-hybridization. Journal of clinical microbiology 33:1292-1295.
Su L. and Chiu C. (2007) Salmonella: clinical importance and evolution of nomenclature. Chang Gung medical journal 30:210.
Whyte P., Mc Gill K., Collins J. and Gormley E. (2002) The prevalence and PCR detection of Salmonella contamination in raw poultry. Veterinary Microbiology 89:53-60.
Widjojoatmodjo M., Fluit A., Torensma R., Verdonk G.and Verhoef J. (1992) The magnetic immunopolymerase chain reaction assay for direct detectionof salmonellae in fecal samples. Journal of clinicalmicrobiology 30:3195-3199.