Volume 3, Issue 4 (December 2016)                   J. Food Qual. Hazards Control 2016, 3(4): 128-133 | Back to browse issues page

XML Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Eslami G, Manafi L, Peletto S. DNA Extraction from Beef Harboring Sarcocystis spp.: Comparison of Three Different Analytical Methods. J. Food Qual. Hazards Control 2016; 3 (4) :128-133
URL: http://jfqhc.ssu.ac.ir/article-1-287-en.html
DNA Extraction from Beef Harboring Sarcocystis spp.: Comparison of Three Different Analytical Methods
G. Eslami , L. Manafi , S. Peletto *
Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d'Aosta, Turin, Italy , simone.peletto@izsto.it
Abstract:   (5811 Views)

Background: DNA extraction is one the most important steps for molecular analysis of food-borne pathogens. In this research, three methods of DNA extractions from beef harboring Sarcocystis spp. were compared for the quality, quantity, safety, as well as cost-effectiveness.

Methods: About 100 mg intersostal and diaphragm were collected from 10 slaughtered cattle. After ensuring their contamination with Sarcocyst using Polymerase Chain Reaction (PCR) with the specific primer pair, three methods of salting out, Phenol-Chloroform-Isoamylalcohol (PCI), and commercial kit were performed. Quantification, qualification, and amplification analysis of the extracted DNA was done using spectrophotometer, agarose gel electrophoresis, and PCR, respectively. Statistical analysis was performed using ANOVA test, by SPSS, Inc, Chicago, IL software (v.16.0).

Results: Qualification in all methods was appropriate but the ones related to salting out and PCI methods were the best in comparison with the ones from commercial kit. Quantification analysis indicated the mean concentration of 249.3±3.94, 67.8±5.1, and 31±2.7 ng/μl for PCI, salting out, and commercial kit, respectively. The purification analysis represented the mean ratios of A (260)/(280), 1.7±0.3, 1.63±0.2, and 1.81±0.6 for PCI, salting out, and commercial kit, respectively. No significant difference (p>0.05) was found between the yielded concentration and purification among three methods.

Conclusion: The commercial kit is expensive, but salting out and PCI methods are cost effectiveness, however the last is considered as a toxic method. Because, amplification in all methods was appropriate, we introduced salting out for molecular detection of Sarcocystis in beef.

Keywords: Sarcocystis, Meat, Molecular Diagnostic Techniques
Full-Text [PDF 370 kb]   (2131 Downloads)    
Type of Study: Original article | Subject: Special
Received: 16/07/12 | Accepted: 16/09/07 | Published: 16/12/14
References
1. Asadzadeh N., Javanmard A., Nassiry M.R. (2010). Comparison of rapid DNA extraction techniques for conventional PCR-RFLP analysis from mammalian whole blood cells. Journal of Molecular Genetics. 2: 32-35.
2. Bhaganna P., Volkers R.J., Bell A.N., Kluge K., Timson D.J., McGrath J.W., Ruijssenaars H.J., Hallsworth J.E. (2010). Hydrophobic substances induce water stress in microbial cells. Microbial Biotechnology. 3: 701-716.
3. Boom R.C.J.A., Sol C.J., Salimans M.M., Jansen C.L., Wertheimvan Dillen P.M., Van Der Noordaa J.P.M.E. (1990). Rapid and simple method for purification of nucleic acid. Journal of Clinical Microbiology. 28: 495-503.
4. Boughattas S., Salehi R. (2014). Molecular approaches for detection and identification of foodborne pathogens. Journal of Food Quality and Hazards Control. 1: 1-6.
5. Davoudi A., Tarang A., Aleyasin S.A., Salehi A., Seighalani R., Tahmoressi F. (2012). Evaluation of two DNA extraction methods from maternal plasma for using in non-invasive bovine fetus gender determination. Iranian Journal of Reproductive Medicine. 10: 523-530.
6. Eaglestone S.S., Ruddock L.W., Cox B.S., Tuite M.F. (2000). Guanidine hydrochloride blocks a critical step in the propagation of the prion-like determinant [PSI+] of Saccharomyces cerevisiae. Iranian Journal of Reproductive Medicine. 97: 240-244.
7. Eslami G., Frikha F., Salehi R., Khamesipour A., Hejazi H., Nilforoushzadeh M.A. (2011). Cloning, expression and dynamic simulation of TRYP6 from Leishmania major (MRHO/IR/75/ER). Molecular Biology Reports. 38: 3765- 3776.
8. Fayer R., Esposito D.H., Dubey J.P. (2015). Human infections with Sarcocystis species. Clinical Microbiology Reviews. 28: 295- 311.
9. Freschi C.R., de Oliveira e Silva Carvalho L.F., de Oliveira C.J.B. (2005). Comparison of DNA-extraction methods and selective enrichment broths on the detection of Salmonella typhimurium in swine feces by polymerase chain reaction (PCR). Brazilian Journal of Microbioliogy. 36: 363-367.
10. Ghisleni G., Robba S., Germani O., Scanziani E. (2006). Identification and prevalence of Sarcocystis spp. cysts in bovine canned meat. Food Control. 17: 691-694.
11. Hajimohammadi B., Eslami G., Oryan A., Zohtabar A., Pourmirzaei Tafti H., Moghaddam Ahmadi M. (2014). Molecular identification of Sarcocystis hominis in native cattle of central Iran: a case report. Tropical Biomedicine. 31: 183-186.
12. Javadi A., Shamaei M., Ziazi L.M., Pourabdollah M., Dorudinia A., Seyedmehdi S.M., Karimi S. (2014). Qualification study of two genomic DNA extraction methods in different clinical samples. Tanaffos. 13: 41-47.
13. Kim Y.C., Morrison S.L. (2009). A rapid and economic in-house DNA purification method using glass syringe filters. PLoS One. 4: e7750.
14. Lee N., Kwon K.Y., Oh S.K., Chang H.J., Chun H.S., Choi S.W. (2014). A multiplex PCR assay for simultaneous detection of Escherichia coli O157:H7, Bacillus cereus, Vibrio parahaemolyticus, Salmonella spp., Listeria monocytogenes, and Staphylococcus aureus in Korea ready-to-eat food. Foodborne Pathogens and Disease. 11: 574-580.
15. Maurya R., Kumar B., Sundar S. (2013). Evaluation of salt-out method for the isolation of DNA from whole blood: a pathological approach of DNA based diagnosis. International Journal of Life Sciences Biotechnology and Pharma Research. 2:53-57.
16. Mirmomeni M.H., Sajjadi Majd S., Sisakhtnezhad S., Doranegard F. (2010). Comparison of the three methods for DNA extraction from paraffin-embedded tissues. Journal of Biological Sciences. 10: 261-266.
17. Oliver S.P., Jayarao B.M., Almeida R.A. (2005). Foodborne pathogens in milk and the dairy farm environment: food safety and public health implications. Foodborne Pathogens and Disease. 2: 115-129.
18. Omurtag I., Paulsen P., Hilbert F., Smulders F.J.M. (2013). The risk of transfer of foodborne bacterial hazards in Turkey through the consumption of meat; risk ranking of muscle foods with the potential to transfer Campylobacter spp. Food Security. 5: 117-127.
19. Rosec J.P., Causse V., Cruz B., Rauzier J., Carnat L. (2012). The international standard ISO/TS 21872-1 to study the occurrence of total and pathogenic Vibrio parahaemolyticus and Vibrio cholerae in seafood: ITS improvement by use of a chromogenic medium and PCR. International Journal of Food Microbiology. 157: 189-194.
20. Sambrook J., Russell D.W. (2006a). Purification of Nucleic Acids by Extraction with Phenol: Chloroform. Cold Spring Harbor Protocols. doi: 10.1101/pdb. prot4455.
21. Sambrook J., Russell D.W. (2006b). Transcriptional Run-on Assays. Cold Spring Harbor Protocols. doi: 10.1101/pdb. prot3956.
22. Tan S.C., Yiap B.C. (2009). DNA, RNA, and protein extraction: the past and the present. BioMed Research International. pp. 1-10.
23. Van de Venter T. (2000). Emerging food-borne diseases: a global responsibility. Food Nutrition and Agriculture. 26: 4- 13.
24. Vangee L., Houf K., Chiers K., Vercruysse J., D'Herde K., Ducatelle R. (2007). Molecular-based identification of Sarcocystis hominis in Belgian minced beef. Journal of Food Protection. 70: 1523-1526.
25. Wingstrand A., Neimann J., Engberg J., Nielsen E.M., Gerner- Smidt P., Wegener H.C., Molbak K. (2006). Fresh chicken as main risk factor for campylobacteriosis, Denmark. Emerging Infectious Diseases. 12: 280-284.
26. Zhao X., Lin C.W., Wang J., Oh D.H. (2014). Advances in rapid detection methods for foodborne pathogens. Journal of Microbiology and Biotechnology. 24: 297-312.
Add your comments about this article : Your username or Email:
CAPTCHA
Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

© 2024 CC BY-NC 4.0 | Journal of food quality and hazards control

Designed & Developed by : Yektaweb