JBM  Vol.6 No.11 , November 2018
vicK Gene as Potential Identification Marker for Staphylococcus aureus
Staphylococcus aureus is an important human pathogen frequently detected in hospital community and has emerged as an important health concern in human medicine. Identification of S. aureus from clinical specimens by phenotypic methods may produce variable characteristics leading to ambiguity. Hence, a rapid and reliable method for identification of S. aureus is required which could expedite appropriate antibiotic therapy. This study aimed to evaluate the specificity of polymerase chain reaction (PCR) targeting a signal transduction gene, vicK, among S. aureus isolates of Hospital Sultanah Nur Zahirah, Kuala Terengganu, Malaysia. A total of 118 bacterial isolates were screened, which consisted of one hundred S. aureus isolates, ten Staphylococcus spp. and eight non-Staphylococci. Results indicated that PCR targeting vicK was able to identify 98% of S. aureus isolates with high sensitivity and specificity, while the remaining isolates of Staphylococcus spp. and non-Staphylococci did not yield any amplification of the gene. vicK thus, is highly specific within interspecies and intraspecies, which is potential to be used as a molecular identification marker for S. aureus.
Cite this paper: Morad, N. , Misbah, S. (2018) vicK Gene as Potential Identification Marker for Staphylococcus aureus. Journal of Biosciences and Medicines, 6, 98-110. doi: 10.4236/jbm.2018.611010.

[1]   Köck, R., Werner, P., Friedrich, A.W., Fegeler, C., Becker, K., Bindewald, O., von Auenmüller, L., et al. (2016) Persistence of Nasal Colonization with Human Pathogenic Bacteria and Associated Antimicrobial Resistance in the German General Population. New Microbes and New Infections, 9, 24-34.

[2]   Becker, K. (2018) Pathogenesis of Staphylococcus aureus. In: Staphylococcus aureus, Elsevier, 13-38.

[3]   Tong, S.Y.C., Davis, J.S., Eichenberger, E., Holland, T.L. and Fowler Jr., V.G. (2015) Staphylococcus aureus Infections: Epidemiology, Pathophysiology, Clinical Manifestations, and Management. Clinical Microbiology Reviews, 28, 603-661.

[4]   Becker, K., Heilmann, C. and Peters, G. (2014) Coagulase-Negative Staphylococci. Clinical Microbiology Reviews, 27, 870-926.

[5]   Lakhundi, S. and Zhang, K. (2018) Methicillin-Resistant Staphylococcus aureus: Molecular Characterization, Evolution, and Epidemiology. Clinical Microbiology Reviews, 31, Article ID: e00020-18.

[6]   Marra, A.R., Camargo, L.F.A., Pignatari, A.C.C., Sukiennik, T., Behar, P.R.P., Medeiros, E.A.S., et al. (2011) Nosocomial Bloodstream Infections in Brazilian Hospitals: Analysis of 2,563 Cases from a Prospective Nationwide Surveillance Study. Journal of Clinical Microbiology, 49, 1866-1871.

[7]   Köck, R., Becker, K., Cookson, B., van Gemert-Pijnen, J.E., Harbarth, S., Kluytmans, J., Friedrich, A.W., et al. (2010) Methicillin-Resistant Staphylococcus aureus (MRSA): Burden of Disease and Control Challenges in Europe. Eurosurveillance, 15, Article ID: 19688.

[8]   Sahibzada, S., Abraham, S., Coombs, G.W., Pang, S., Hernández-Jover, M., Jordan, D. and Heller, J. (2017) Transmission of Highly Virulent Community-Associated MRSA ST93 and Livestock-Associated MRSA ST398 between Humans and Pigs in Australia. Scientific Reports, 7, 5273.

[9]   Cuny, C., Wieler, L.H. and Witte, W. (2015) Livestock-Associated MRSA: The Impact on Humans. Antibiotics (Basel, Switzerland), 4, 521-543.

[10]   DeLeo, F.R., Otto, M., Kreiswirth, B.N. and Chambers, H.F. (2010) Community-Associated Meticillin-Resistant Staphylococcus aureus. Lancet (London, England), 375, 1557-1568.

[11]   de LourdesRibeiro de Souza da Cunha, M. (2018) Methods for the Identification, Characterization, and Tracking the Spread of Staphylococcus aureus. In: Staphylococcus aureus, Elsevier, 105-125.

[12]   Brown, D.F.J., Edwards, D.I., Hawkey, P.M., Morrison, D., Ridgway, G.L., Towner, K.J. and Wren, M.W.D. (2005) Guidelines for the Laboratory Diagnosis and Susceptibility Testing of Methicillin-Resistant Staphylococcus aureus (MRSA). Journal of Antimicrobial Chemotherapy, 56, 1000-1018.

[13]   Shittu, A., Lin, J., Morrison, D. and Kolawole, D. (2006) Identification and Molecular Characterization of Mannitol Salt Positive, Coagulase-Negative Staphylococci from Nasal Samples of Medical Personnel and Students. Journal of Medical Microbiology, 55, 317-324.

[14]   Kawamura, Y., Hou, X.G., Sultana, F., Hirose, K., Miyake, M., Shu, S.E. and Ezaki, T. (1998) Distribution of Staphylococcus Species among Human Clinical Specimens and Emended Description of Staphylococcus caprae. Journal of Clinical Microbiology, 36, 2038-2042.

[15]   Kateete, D.P., Kimani, C.N., Katabazi, F.A., Okeng, A., Okee, M.S., Nanteza, A., Najjuka, F.C., et al. (2010) Identification of Staphylococcus aureus: DNase and Mannitol Salt Agar Improve the Efficiency of the Tube Coagulase Test. Annals of Clinical Microbiology and Antimicrobials, 9, 23.

[16]   Qian, Q., Eichelberger, K. and Kirby, J.E. (2007) Rapid Identification of Staphylococcus aureus in Blood Cultures by Use of the Direct Tube Coagulase Test. Journal of Clinical Microbiology, 45, 2267-2269.

[17]   Saraiva, M.M., De Leon, C.M., Santos, S.C., Stipp, D.T., Souza, M.M., Santos Filho, L., Oliveira, C.J., et al. (2018) Accuracy of PCR Targeting Different Markers for Staphylococcus aureus Identification: A Comparative Study Using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry as the Gold Standard. Journal of Veteri-nary Diagnostic Investigation, 30, 252-255.

[18]   Ghodousi, A., Nomanpour, B., Davoudi, S., Maleknejad, P., Omrani, M., Kashef, N., Feizabadi, M.M., et al. (2012) Application of fnbA Gene as New Target for the Species-Specific and Quantitative Detection of Staphylococcus aureus Directly from Lower Respiratory Tract Specimens by Real Time PCR. Indian Journal of Pathology & Microbiology, 55, 490-495.

[19]   Wu, Q., Li, Y., Hu, H., Wang, M., Wu, Z. and Xu, W. (2012) Rapid Identification of Staphylococcus aureus: FISH versus PCR Methods. Laboratory Medicine, 43, 276-280.

[20]   Yang, Y.-C., Yu, H., Xiao, D.-W., Liu, H., Hu, Q., Huang, B., Huang, W.-F., et al. (2009). Rapid Identification of Staphylococcus aureus by Surface Enhanced Laser Desorption and Ionization Time of Flight Mass Spectrometry. Journal of Microbiological Methods, 77, 202-206.

[21]   Spence, R.P., Wright, V., Ala-Aldeen, D.A.A., Turner, D.P., Wooldridge, K.G. and James, R. (2008) Validation of Virulence and Epidemiology DNA Microarray for Identification and Characterization of Staphylococcus aureus Isolates. Journal of Clinical Microbiology, 46, 1620-1627.

[22]   Misbah, S., Hassan, H., Yusof, M.Y., Hanifah, Y.A. and Abubakar, S. (2005) Genomic Species Identification of Acinetobacter of Clinical Isolates by 16S rDNA Sequencing Genomic Species Identification of Acinetobacter of Clinical Isolates by 16S rDNA Sequencing. Singapore Medical Journal, 46, 461-464.

[23]   Riyaz-Ul-Hassan, S., Verma, V. and Qazi, G.N. (2008) Evaluation of Three Different Molecular Markers for the Detection of Staphylococcus aureus by Polymerase Chain Reaction. Food Microbiology, 25, 452-459.

[24]   Tiwari, H.K., Sapkota, D. and Sen, M.R. (2008) Evaluation of Different Tests for Detection of Staphylococcus aureus Using Coagulase (coa) Gene PCR as the Gold Standard. Nepal Medical College Journal: NMCJ, 10, 129-131.

[25]   Poyart, C., Quesne, G., Boumaila, C. and Trieu-Cuot, P. (2001) Rapid and Accurate Species-Level Identification of Coagulase-Negative Staphylococci by Using the sodA Gene as a Target. Journal of Clinical Microbiology, 39, 4296-4301.

[26]   Brakstad, O.G., Aasbakk, K. and Maeland, J.A. (1992) Detection of Staphylococcus aureus by Polymerase Chain Reaction Amplification of the nuc Gene. Journal of Clinical Microbiology, 30, 1654-1660.

[27]   Cremonesi, P., Perez, G., Pisoni, G., Moroni, P., Morandi, S., Luzzana, M., Castiglioni, B., et al. (2007) Detection of Enterotoxigenic Staphylococcus aureus Isolates in Raw Milk Cheese. Letters in Applied Microbiology, 45, 586-591.

[28]   Zhang, K., Sparling, J., Chow, B.L., Elsayed, S., Hussain, Z., Church, D.L., Conly, J.M., et al. (2004) New Quadriplex PCR Assay for Detection of Methicillin and Mupirocin Resistance and Simultaneous Discrimination of Staphylococcus aureus from Coagulase-Negative Staphylococci. Journal of Clinical Microbiology, 42, 4947-4955.

[29]   Chikkala, R., Oommen George, N., Ratnakar, K.S., Natarajan Iyer, R. and Sritharan, V. (2012) Heterogeneity in femA in the Indian Isolates of Staphylococcus aureus Limits Its Usefulness as a Species Specific Mark. Advances in Infectious Diseases, 2, 82-88.

[30]   Schwarzkopf, A. and Karch, H. (1994) Genetic Variation in Staphylococcus aureus Coagulase Genes: Potential and Limits for Use as Epidemiological Marker. Journal of Clinical Microbiology, 32, 2407-2412.

[31]   Liu, Z., Zhou, Z., Huang, Y. and Chen, R. (2017) Molecular Identification of Staphylococcus aureus Isolated from Clinical Samples by Specific PCR Assay Targeting the Signal Transduction Gene. Journal of Pharmacy and Pharmaceutics, 4, 166-171.

[32]   Liu, Z., Shi, X. and Pan, F. (2007) Species-Specific Diagnostic Marker for Rapid Identification of Staphylococcus aureus. Diagnostic Microbiology and Infectious Disease, 59, 379-382.

[33]   Mattos-Graner, R.O. and Duncan, M.J. (2017) Two-Component Signal Transduction Systems in Oral Bacteria. Journal of Oral Microbiology, 9, Article ID: 1400858.

[34]   Dashti, A., Dashti, H. and Jadaon, M. (2009) Heat Treatment of Bacteria: A Simple Method of DNA Extraction for Molecular Techniques. Journal of the Kuwait Medical Association.

[35]   Gitau, W., Masika, M., Musyoki, M., Museve, B. and Mutwiri, T. (2018) Antimicrobial Susceptibility Pattern of Staphylococcus aureus Isolates from Clinical Specimens at Kenyatta National Hospital. BMC Research Notes, 11, 226.

[36]   Wankhade, A.B., Panda, S., Hathiwala, R. and Keche, Y. (2017) Study of Antibiotic Resistance Profiling of Staphylococcus aureus Isolated from Clinical Specimens of the Patients Attending a Tertiary Teaching Hospital from Chhattisgarh. International Journal of Research in Medical Sciences, 5, 4808.

[37]   Bindu, D., Saikumar, C. and Vinith, M. (2017) Prevalence and Antibiotic Susceptibility of Methicillin Resistant Staphylococcus aureus Isolates in a Tertiary Care Centre. Journal of Pharmaceutical Sciences and Research, 9, 2329-2331.

[38]   Kobayashi, S.D., Malachowa, N. and DeLeo, F.R. (2015) Pathogenesis of Staphylococcus aureus Abscesses. The American Journal of Pathology, 185, 1518.

[39]   Foster, T. (1996) Staphylococcus. In: Medical Microbiology, University of Texas Medical Branch at Galveston.

[40]   Shuping, L.L., Kuonza, L., Musekiwa, A., Iyaloo, S. and Perovic, O. (2017) Hospital-Associated Methicillin-Resistant Staphylococcus aureus: A Cross-Sectional Analysis of Risk Factors in South African Tertiary Public Hospitals. PLOS ONE, 12, e0188216.

[41]   Carnicer-Pont, D., Bailey, K.A., Mason, B.W., Walker, A.M., Evans, M.R. and Salmon, R.L. (2006) Risk Factors for Hospital-Acquired Methicillin-Resistant Staphylococcus aureus Bacteraemia: A Case-Control Study. Epidemiology and Infection, 134, 1167.

[42]   Muenks, C.E., Hogan, P.G., Wang, J.W., Eisenstein, K.A., Burnham, C.-A.D. and Fritz, S.A. (2016) Diversity of Staphylococcus aureus Strains Colonizing Various Niches of the Human Body. The Journal of Infection, 72, 698-705.

[43]   Paterson, G.K., Harrison, E.M., Murray, G.G.R., Welch, J.J., Warland, J.H., Holden, M.T.G., Holmes, M.A., et al. (2015) Capturing the Cloud of Diversity Reveals Complexity and Heterogeneity of MRSA Carriage, Infection and Transmission. Nature Communications, 6, 6560.

[44]   Lalonde, L.F. and Gajadhar, A.A. (2008) Highly Sensitive and Specific PCR Assay for Reliable Detection of Cyclospora cayetanensis Oocysts. Applied and Environmental Microbiology, 74, 4354-4358.

[45]   Siqueira, J.F. and Rôças, I.N. (2003) PCR Methodology as a Valuable Tool for Identification of Endodontic Pathogens. Journal of Dentistry, 31, 333-339.

[46]   Liu, Y., Zhang, J. and Ji, Y. (2016) PCR-Based Approaches for the Detection of Clinical Methicillin-Resistant Staphylococcus aureus. The Open Microbiology Journal, 10, 45-56.

[47]   Abolmaaty, A., El-Shemy, M.G., Khallaf, M.F. and Levin, R.E. (1998) Effect of Lysing Methods and Their Variables on the Yield of Escherichia coli O157: H7 DNA and Its PCR Amplification. Journal of Microbiological Methods, 34, 133-141.

[48]   Heikens, E., Fleer, A., Paauw, A., Florijn, A. and Fluit, A.C. (2005) Comparison of Genotypic and Phenotypic Methods for Species-Level Identification of Clinical Isolates of Coagulase-Negative Staphylococci. Journal of Clinical Microbiology, 43, 2286-2290.

[49]   Ayeni, F.A. and Odumosu, B.T. (2016) False Identification of Other Microorganisms as Staphylococcus aureus in Southern Nigeria. Tropical Journal of Pharmaceutical Research, 15, 1941.

[50]   Ahmed, M.O., Abuzweda, A.R., Alghazali, M.H., Elramalli, A.K., Amri, S.G., Aghila, E.S. and Abouzeed, Y.M. (2010) Misidentification of Methicillin-Resistant Staphylococcus aureus (MRSA) in Hospitals in Tripoli, Libya. The Libyan Journal of Medicine, 5.

[51]   Leung, M.J., Nuttall, N., Mazur, M., Taddei, T.L., McComish, M. and Pearman, J.W. (1999) Case of Staphylococcus schleiferi Endocarditis and a Simple Scheme to Identify Clumping Factor-Positive Staphylococci. Journal of Clinical Microbiology, 37, 3353-3356.

[52]   Freney, J., Brun, Y., Bes, M., Meugnier, H., Grimont, F., Grimont, P.A.D., Fleurette, J., et al. (1988) Staphylococcus lugdunensis sp. nov. and Staphylococcus schleiferi sp. nov., Two Species from Human Clinical Specimens. International Journal of Systematic Bacteriology, 38, 168-172.