Characterization and Detection of mecA gene in Different Species of Staphylococcus, Streptococcus and Kocuria Which Isolated from Thalassemia Patients in Erbil City

Related Articles

Special Edition No2 2017 Cihan University-Erbil Scientific Journal

August 2, 2017

Cytogenetic Studies on Lymphocytes Isolated from Women with a History of Abortion Infected with Toxoplasma gondii

August 2, 2017

Investigation of Microsatellite Instability BAT 25 and BAT 26 in Breast Cancer Patients by Conventional PCR

August 2, 2017

Author: Zheen A. Abdulla
Department of Pharmacognosy, College of Pharmacy, University of Hawler medical

Author: Khadija Kh. M. Barzani
Department of Biology, College of Education, University of Salahaddin -Erbil,

DOI: http://dx.doi.org/10.24086/cuesj.si.2017.n2a8

Abstract

Out of 96specimens; 52 oral cavity swabs and 44 urine samples were collected from thalassemic patients in Erbil city . The results showed that 32 isolates of Gram positive bacteria were identified depending on VITEK 2 compact system . On the other hand, the results  showed that 3 genera and 14species of  Gram positive bacteria were isolated and among the isolates the predominant obtained bacteria was streptococci which 13(40.625%) isolates belong to 6 species, followed by Staphylococci which 10 (31.25%) isolates belong to 5 species and  9 (9.375%) isolates of kocuria belonged to 3 species .  Some virulence factors such as oxidase, catalase, coagulase, DNase, protease, lecithinase, lipase, gelatinase, urease, hemolysin and β-lactamase were studied and the results showed that the isolated bacteria were differ in their ability to produce these above mentioned enzymes. All isolated bacteria were tested for their ability for biofilm formation using microtiter plate assay and the results revealed that all isolates were biofilm producer. The susceptibility of bacterial isolates against 10 antibiotics was tested and it was found that the most of the isolated species were highly resistant methicillin with percentage 71.19% , followed by  cloxacillin  and tetracycline  with percentage 52.74% and 52.5% respectively, and also it was showed variable resistance to other antibiotics. Another part of the study concerned with using polymerase chain reaction technique for detection of mecA gene .The results indicated that all isolated Staphylococci speceis were harboring mecA gene with the product size 533 bp except S. vitulinus. . Also was  found mecA gene in one isolate of K.rosea with same product size of above mentioned mecA, whereas no genes were obtained from other isolated bacteria and the detected gene of mecA in S.aureus and K.rosea was sequenced to confirm the results of mecA gene.

Keywords:  Staphylococci, Streptococci, Kocuria, enzymes, biofilm, antibiotics and mecA gene

References

• Saud, AM. ; Al-Azzawie, HF.; & Al-Kazaz, AA. . Molecular Study on β-Thalassemia Patients in Iraq. Current Research in Microbiology and Biotechnology. 1(4) ,2013, 160-165.
• Madhok, S. & Madhok, S. .Dental considerations in Thalassemia patients. Journal of Dental and Medical Sciences. 13(6) ,2014, p.57-62.
• Teawtrakul, N.; Jetsrisuparb, A.; Sirijerachai, C.; Chanung, K. & Wanitpongpun, C. . Severe bacterial infections in patients with non-transfusion- dependent thalassemia: prevalence and clinical risk factors. International Journal of Infectious Diseases. 39,2015, P. 53–56.
• Vento, S. & Cainelli. Infections and thalassaemia. The Lancet Infectious Diseases. 6 (4) ,2016, p. 226–233.
• Cappellini, MD.; Cohen, A.; Porter, J.; Taher, A. & Viprakasit, V. Guidelines for the management of transfusion dependent thalassaemia (tdt). 3^rd Team up Creations Ltd. Nicosia, Cyprus, 2014.
• Soto, S. M. Role of efux pumps in the antibiotic resistance of bacteria embedded in a bioflm. Landes Bioscience. 4 (3),2013, .p. 1-7.
• Adwan, G.; Adwan, K.; Jarrar, N. & Amleh, A. Molecular detection of nine antibiotic resistance genes in methicillin resistant staphylococcus aureus Romanian Archives of Microbiology and Immunology. 73(1-2), 2014, p.9-18.
• Abbadi, S.; Youssef, H.; Nemenqani, D. & Abdel- Moneim, A.S. Rapid identification of methicillin resistant Staphylococcus aureus using real time PCR. Advances in Infectious Diseases. 3, 2013, 44-49.
• Khan, S. AK.; Shetty, P.J.; Sarayu, L.Y.; Chidambaram, A. & RanganathanN, R. Detection of mecA genes of Methicillin-Resistant Staphylococcus aureus by Polymerase Chain Reaction. International Journal of Health and Rehabilitation Sciences. 1(2), 2012, p. 64-68.
• Pincus, D.H. Microbial Identification using the biomerieux VITEK2 system. American Society for Microbiology, Washington, D.C. USA ,2010.
• Sharma, K . Manual of MICROBIOLOGY tools & Techniques. 2^nd Ane books pvt. Ltd. Newdelhi, India , 2007.
• Cappuccino, J.G. & Sherman, NMicrobiology: A laboratory Manual. 8^th Peason Benjamin Cummings. San Francisco, U.S. ,2008.
• Jarjees, K.K. . spa gene and AntibioGram Typing of Staphylococcus aureus isolated from different clinical sources in Erbil city. PhD. Thesis. College of Education- University of Salahaddin-Erbil- Iraq, 2014.
• Stratev, D.; Daskalov, H. & Vashin, I . Characterisation and determination of antimicrobial resistance of b-haemolytic Aeromonas Isolated from common carp (Cyprinus carpio L.). Revue de médecine vétérinaire. 166 (1-2) , 2015, p. 54-61.
• Mathur, T.; Singhal, S.; Khan, S.; Upadhyay, D.J.; Fatma, T. & Rattan, A. Detection of Biofilm Formation Among the Clinical Isolates of Staphylococci: an Evaluation of Three Different Screening Methods. Indian Journal of Medical Microbiology. 24(1), 2006,  25-29.
• Clinical and Laboratory Standard Institute. Performance Standards for Antimicrobial Susceptibility Testing; Twenty-First Informational Supplement Clinical and Laboratory Standards Institute antimicrobial susceptibility testing standards, CLSI 31- M100-S16. ,2011.
• Kareem, N.H. Detection and Evaluation of Methicillin-Resistant Staphylococcus aureus by Duplex PCR. Journal of Al-Nahrain University Science. 16 (2) , 2013, p.157-162.
• Igeltjorn, L. Molecular and biochemical characterization of naturally occurring hyper expressed and mutated extended spectrum AmpC β-lactamases in norwegian clinical isolates of Escherichia coli. MSc. Thesis. University of Tromso, 2009)
• Mishra, V.; Nag, V.L.; Tendon, R. & Awsthi, SResponse surface methodology- Based optimization of agarose gel electrophoresis for screening and electropherotyping of Rotavirus. Applied Biochemistry and Biotechnology. 160(8) , 2010, 2322-2331.
• Stipcevic, T.; Piljac, T. & Isseroff, R. R. Di-rhamnolipid from Pseudomonas aeruginosa displays differential effects on human keratinocyte and fibroblast cultures. Journal of Dermatological Science. 40 (2) , 2005, p. 141-143.
• Ryan, K. J. & Ray, C. G. SHERRIS MEDICAL MICROBIOLOGY (AN INTRODUCTION TO INFECTIOUS DISEASES). 4^th MCGRAW-HILL Companies, Inc. USA, 2004.
• Savini, V.; Catavitello, C.; Masciarelli, G.; Astolfi, D.; Balainot, A.; Bianco, A.; FEBBO, F.; Damario, C. & Dantonio, D. Drug sensitivity and clinical impact of members of the genus Kocuria. Journal of Medical Microbiology. 59, 2010,  .p. 1395–1402.
• Deka, N. Comparison of Tissue Culture plate method, Tube Method and Congo Red Agar Method for the detection of biofilm formation by Coagulase Negative Staphylococcus isolated from Non-clinical Isolates. International Journal of Current Microbiology and Applied Sciences. 3(10) ,2014 .p. 810-815.
• Stepanovic, S.; Vukovic, D.; Dakic, I.; Savic, B. & Svabic- Vlahovic, M. A modified microtiter-plate test for quantification of staphylococcal biofilm formation. Journal of Microbiological Methods. 40 , 2000, p. 175-179.
• Lemaire, S.; Bambeke, F. V.; Mingeot-Leclerco, MP.; Glupczynski, Y. & Tulkens, P. M. Role of acidic pH in the susceptibility of intraphagocytic methicillin-resistant Staphylococcus aureus strains to meropenem and cloxacillin. Atimicrobial agents and Chemotherapy. 51(5) , 2007, p. 1627–1632.
• Duran, N.; Ozer, B.; Duran, G.G.; Onlen, Y. & Demir, C. Antibiotic resistance genes & susceptibility patterns in Staphylococci. Indian Journal of Medical Research. 135, 2012, 389-396.
• Parashar, S. Staphylococcus Haemolyticus; A nosocomial pathogen showing higher antimicrobial resistance. International journal of scientific research. 3 (1) , 2014, 381-382.
• AlL-Marzoqi, A. H.; Shemmran, A. R.; Al-Hindi, Z. SH.; Al-Taee, M. & Al-Nafee, M. K . Bacterial and Viral Infections associated with Thalassemia Patients in Hillah City. Journal of Al-Qadisiyah for pure Science. 14 (3), 2009.
• Morera, J.S.; Riccetto, A.G.; DailvaA, M.T. & Vilela, M.M. Endocarditis by Kocuria rosea in an immunocompetent child. The Brazilian Journal of infectious diseases. 9 (1) , 2015,  82–84.
• Onwubiko, N. E. & Sadiq, N. M. Antibiotic sensitivity pattern of Staphylococcus aureus from clinical isolates in a tertiary health institution in Kano, Northwestern Nigeria. Pan African Medical Journal. 8 (4) , 2011, p. 1-7.
• Huang, ZG.; Zheng, XZ.; Guan, J.; Xiao, SN. & Zhuo, C. Direct Detection of Methicillin-Resistant Staphylococcus aureus in Sputum Specimens from Patients with hospital-associated pneumonia using a Novel Multilocus PCR Assay. Pathogens. 4 , 2015, p. 199-209.
• Aziz, H.W.; Al-Dulaimi, T.H.; Al-Marzoqi, A.H. & Ahmed, N.K. Phenotypic detection of resistance in Staphylococcus aureus isolates: Detection of (mecA and femA) gene in methicillin resistant Staphylococcus aureus (MRSA) by Polymerase Chain Journal of Natural Sciences Research. 4(1), 2014, p.112-118.
• S.; Gwarzo, Y. & Uba, A . One step PCR for detection of Staphylococcus aureus specific sequence gene and mecA Gene. International Journal of Advanced Materials Research. 1(3) , 2015, p. 73-79.