Identification of Methicillin resistant Staphylococcus aureus in yoghurts from Şanlıurfa by real-time PCR

Abstract

Methicillin resistant Staphylococcus aureus (MRSA) is one of the most important pathogenic microorganisms that display multiple drug 

resistance, causing severe infections in animals and humans. Contaminated or raw milk products, such as yoghurt, are one of the root causes

of such MRSA infections for the consumers. Recent studies on clonal investigation revealed the same clonal isolates of animals in human

infections. Thus, the epidemiology of these bacteria in food products and animals has gained importance. Therefore, the aim of this study was

to epidemiologically detect MRSA in yoghurt samples from Urfa via real-time PCR. To detect MRSA via real-time PCR, the mecA gene was

investigated on the DNA obtained from the yoghurt samples. Among 64 yoghurt samples, of which 55 were unpackaged and 9 were packaged,

the MRSA mecA gene was only detected in 2 (3.13 %) unpackaged samples via real-time PCR. These results showed that it was possible to

detect and epidemiologically investigate MRSA, an important pathogen for human and animal health, via real-time PCR from yoghurt samples

with direct DNA isolation. In addition, this pathogen can be found in the products sold in Şanlıurfa, which indicates that it may lead to severe

diseases in case of consumption, if not conformed to hygienic conditions.

Keywords

• MRSA,contamination,real-time PCR,yoghurt

References

1. [1] Köck R, Becker K, Cookson B, van Gemert-Pijnen JE, Harbarth S, Kluytmans J, Mielke M, Peters G, Skov RL, Struelens MJ, Tacconelli E, Navarro Torné A, Witte W, Friedrich AW.2010. Methicillin-resistant Staphylococcus aureus (MRSA): burden of disease and control challenges in Europe. Euro Surveill, 15(41).
2. [2] Graveland H1, Duim B, van Duijkeren E, Heederik D, Wagenaar JA. 2011. Livestock-associated methicillinresistant Staphylococcus aureus in animals and humans. Int J Med Microbiol, 301(8),630-4.
3. [3] Petinaki E, Spiliopoulou I. 2012. Methicillin-resistant Staphylococcus aureus among companion and foodchain animals: impact of human contacts. ClinMicrobiol Infect, 18(7),626-34.
4. [4] Iqbal Z, Seleem MN, Hussain HI, Huang L, Hao H, Yuan Z. 2016. Comparative virulence studies and transcriptome analysis of Staphylococcus aureus strains isolated from animals. Scientific Reports, 6,35442. J Dairy Sci, 99(10), 7872-7876. [5] Lozano C, Gharsa H, BenSlama K, Zarazaga M, Torres C . 2016. Staphylococcus aureus in Animals and Food: Methicillin Resistance, Prevalence and Population Structure. A Review in the African Continent. Poirel L, ed. Microorganisms, 4(1),12.
5. [6] Marshall BM, Levy SB .2011. Food animals and antimicrobials: impacts on human health. Clin. Microbiol. Rev, 24,718–733.
6. [7] Sasidharan S, Prema B, Yoga LL. 2011. Antimicrobial drug resistance of Staphylococcus aureus in dairy products. Asian Pac J Trop Biomed, 1(2):130-132.
7. [8] Ertaş, N, Serhat, AL, Karadal, F,Gönülalan, Z .2014. Kayseri ilinde satışa sunulan manda yoğurtlarının mikrobiyolojik kalitesi. İstanbul ÜniversitesiVeterinerFakültesi- Dergisi, 40(1), 83-89.
8. [9] Galanis A, Kourkoutas Y, Tassou CC, Chorianopoulos N. 2015. Detection and Identification of probiotic Lactobacillus plantarum strains by multiplex PCR using RAPD-Derived primers. Int J MolSci, 16(10), 25141-53.

9. [10] Shrestha NK, Tuohy MJ, Hall GS, Isada CM, Procop GW. 2002. Rapid Identification of Staphylococcus aureus and the mecA Gene from BacT/ALERT blood culture bottles by using the light cycler system. J. Clin. Microbiol, 40(7), 2659-2661.
10. [11] Rohrer S, Tschierske M, Zbinden R, Berger-Bächi B. 2001. Improved methods for detection of Methicillin-Resistant Staphylococcus aureus. Eur J ClinMicrobiol Infect Dis, 20 , 267–270.
11. [12] Grisold AJ, Kessler HH. 2016. Use of Hybridization Probes in a Real-Time PCR Assay on the LightCycler® for the Detection of Methicillin-Resistant Staphylococcus aureus Diagnostic Bacteriology Protocols in Volume 345 of the series Methods in Molecular Biology™ , 79-89. Humana Press.
12. [13] Wada M, Lkhagvadorj E, Bian L, Wang C, Chiba Y, Nagata S, Shimizu T, Yamashiro Y, Asahara T, Nomoto K. 2010. Quantitative reverse transcription-PCR assay for the rapid detection of methicillin-resistant Staphylococcus aureus. J ApplMicrobiol, 108 ,779–788.
13. [14] Zhang Z, Liu W, Xu H, Aguilar ZP, Shah NP, Wei H .2015. Propidiummonoazide combined with real-time PCR for selective detection of viable Staphylococcus aureus in milk powder and meat products. J Dairy Res, 1625–1633.
14. [15] Onen SP, Elmalı M. 2016. Determination of L.monocytogenes, and its Antibiotic Resistance of Local Produced Cheese Consuming in Hatay. Van Vet J, 27(1), 25-29.
15. [16] Al-Ashmawy MA, Sallam KI, Abd-Elghany SM, Elhadidy M, Tamura T. 2016. Prevalence, Molecular Characterization, and Antimicrobial Susceptibility of Methicillin-Resistant Staphylococcus aureus Isolated from Milk and Dairy Products.Foodborne Pathog Dis, 13(3), 156-162.
16. [17] Fadel HM, Ismail J. 2015. Occurrence and zoonotic importance of methicillin-resistant Staphylococcus aureus in raw milk and some dairy products at Ismailia City, Egypt,Hanaa,Zagazig Veterinary Journal (ZVT), 43(3), 95-104.
17. [18] Kamal RM, Bayoumi MA, Abd El Aal SFA. 2013. MRSA detection in raw milk, some dairy products and hands of dairy workers in Egypt, a mini-survey.Food Control, 33(1), 49–53.
18. [19] Rahimi E. 2013. Enterotoxigenicity of Staphylococcus aureus isolated from traditional and commercial dairy products marketed in Iran. Braz J Microbiol , 44 (2), 393-399.
19. [20] Herrera FC, García-López ML, Santos JA. 2016. Characterization of methicillin-resistant Staphylococcus aureus isolated from raw milk fresh cheese in Colombia.
20. [21] Zhang L, Li Y, Bao H, Wei R, Zhou Y, Zhang H, Wang R. 2016. Population structure and antimicrobial profile of Staphylococcus aureus strains associated with bovine mastitis in China. Microb Pathog,103-9.
21. [22] Laurent F, Chardon H, Haenni M, Bes M, Reverdy ME, Madec JY, Lagier E, Vandenesch F, Tristan A. 2012. MRSA harboring mecA variant gene mecC, France.Emerg Infect Dis, 8 (9), 1465.
22. [23] Gezgen C, Seker E. 2016. Investigation of methicillin resistance and Panton-Valentine leukocidin in Staphylococci isolated from bovine mastitis.ActaSci Vet, 44, 01-09.