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Comparative Study of Three β Lactamase Test Methods in Staphylococcus aureus Isolated from Two Nepalese Hospitals

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DOI: 10.4236/ojcd.2014.41009    4,606 Downloads   6,173 Views   Citations

ABSTRACT

Background: β lactamase is a plasmid-encoded enzyme that hydrolyzes β lactam ring of β lactam antibiotics rendering them ineffective. These enzymes, produced by Staphylococcus aureus along with many other organisms, have hindered the use of many useful and once life-saving β lactam antibiotics from clinical practice. Methods: This study was aimed to compare three test methods-chromogenic, acidimetric and iodometric-for the detection of β lactamase enzyme produced by 404 nosocomial induced S. aureus isolated from two Nepali hospitals, Kathmandu based hospital (KBH) and Lalitpur based Hospital (LBH). The study was carried out following standard methodology during November 2007 to June 2009 in the Department of Microbiology, Institute of Medicine, Kathmandu, Nepal. Sensitivity, specificity, efficiency, positive predictive value, and negative predictive values of the tests were calculated taking penicillin resistance and sensitivity as the standard. Results: Chromogenic method was found to be the most sensitive (98.93%) and efficient (98.51%) test and had a high positive predictive value (99.46%). Sensitivity (98.4%) and efficiency (98.27%) of iodometric method was found to be comparable to chromogenic test; its specificity (96.55 %) and positive predictive value (99.73%) were the highest among the 3 tests. Acidimetric test was the least sensitive (97.33%) and efficient (96.78%). Of note, the sensitivity and specificity of these test methods have been compromised due to the negativity of few penicillin resistant isolates and positivity of some penicillin sensitive isolates, respectively. Conclusion: Chromogenic method was found comparatively to be the best test method for the detection of β lactamase production. However, in contrast to the other two test methods whose reagents can be locally and economically prepared, chromogenic test’s use has been impeded by its cost and unavailability in the local Nepali market.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

Bidya, S. and Suman, R. (2014) Comparative Study of Three β Lactamase Test Methods in Staphylococcus aureus Isolated from Two Nepalese Hospitals. Open Journal of Clinical Diagnostics, 4, 47-52. doi: 10.4236/ojcd.2014.41009.

References

[1] Daum, R.S. (1988) Community Acquired Methicillin Resistant Staphylococcal Infections. The Pediatric Infectious Disease Journal, 17, 745-746. http://dx.doi.org/10.1097/00006454-199808000-00015
[2] Barber, M. and Rozwadowska-Dowzenko, M. (1948) Infection by Penicillin Resistant Staphylococci. Lancet, 1, 641-644. http://dx.doi.org/10.1016/S0140-6736(48)92166-7
[3] Jassen, O., Rosendel, K., Bulow, P., Faber, V. and Erikson, K.R. (1969) Changing Staphylococci and Staphylococcal Infections: A Ten Year Study of Bacteria and Cases of Bacterimia. The New England Journal of Medicine, 281, 627-635. http://dx.doi.org/10.1056/NEJM196909182811201
[4] Jevons, M.P. (1961) “Celbenin”—Resistant Staphylococci. British Medical Journal, 1, 124-125. http://dx.doi.org/10.1136/bmj.1.5219.124-a http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1952888/
[5] Hartman, B.S. and Tomasz, A. (1984). A Low Affinity Penicillin Binding Protein Associated in Beta Lactam Resistance in Staphylococcus aureus. Journal of Bacteriology, 158, 513-516.
[6] Hackbarth, C.J. and Chambers, H.F. (1989) Methicillin Resistant Staphylococci: Detection Methods and Treatment of Infections. Antimicrob Agents and Chemother, 33, 995-999. http://dx.doi.org/10.1128/AAC.33.7.995
[7] Hugo, W.B. and Russell, A.D. (1986) Pharmaceutical Microbiology. 3rd Edition, Blackwell Scientific Publications, London.
[8] Song, M.D., Wachi, M., Doi, M., Ishino, F. and Matsuhashi, M. (1987) Evolution of an Inducible Penicillin—Target Protein in Methicillin Resistant Staphylococcus aureus by Gene Fusion. FEBS Letters, 221, 167-171. http://dx.doi.org/10.1016/0014-5793(87)80373-3
[9] Hiramatsu, K. (1995) Molecular Evolution of MRSA. Microbiology and Immunology, 39, 531-543. http://dx.doi.org/10.1111/j.1348-0421.1995.tb02239.x
[10] Joris, B., Hardt, K. and Ghuysen, G.M. (1994) Induction of β Lactamase and Low Affinity Penicillin Binding Protein 2′ Synthesis in Gram Positive Bacteria. In: Ghuysen, J.M. and Hakenbeck, R.M., Eds., Bacterial Cell Wall, Elsevier Science, New York, 505-515.
[11] Chambers, H.F. (1997) Methicillin Resistance in Staphylococci: Molecular and Biochemical Basis and Clinical Implications. Clinical Microbiology Reviews, 10, 781-791.
[12] Isenberg, H.D. (2004) Clinical Microbiology Procedure Handbook. Vol. 1, American Society for Microbiology, ASM Press.
[13] Clinical Laboratory Standards Institute (2007) Performance Standard for Antimicrobial Susceptibility Testing: Seventeenth Informational Supplement M100-S17, Vol. 27, No. 1, Clinical Laboratory Standards Institute, Wayne.
[14] Isenberg, H.D. (2004) Clinical Microbiology Procedure Handbook. Vol. 2, American Society for Microbiology, ASM Press.
[15] Package Insert. Nitrocefin disc, BBL, USA.
[16] Forbes, B.A., Sahm, D.F. and Weissfeld, A. (2007) Bailey and Scott’s Diagnostic Microbiology. 12th Edition, Mosby Elsevier, St Louis, Missouri.
[17] Shrestha, B. (2007) Study of β Lactamase Activity of Staphylococcus aureus Isolated from Healthy Nasal Carriers and Hospital Isolates. Nepal Medical College Journal, 9, 107-110.
[18] Richmond, M.H. (1979) Beta-Lactam Antibiotics and Beta-Lactamases: Two Sides of a Continuing Story. Clinical Infectious Diseases, 1, 30-38. http://dx.doi.org/10.1093/clinids/1.1.30
[19] Samant, S.A. and PAI, C. (2012) Comparative Evaluation of β Lactamase Detection Methods in Staphylococci. International Journal of Research in Pharmaceutical and Biomedical Sciences, 3, 1580-1588.
[20] Oxoid. Nitrocefin (Glaxo Research 87/312). http://www.oxoid.com/UK/blue/index.asp?c=UK&lang=EN
[21] Yu, C., Karr, T., Turner, D., Towns, V. and Sinha, J. (1999) Rapid Detection of Beta-Lactamase Production in Penicillin Sensitive Staphylococci by the Phoenix? Automated ID/AST System. The European Congress of Clinical Microbiology and Infectious Disease (ECCMID), March 1999, Berlin.
[22] Petersson, A.C., Eliasson, I., Kamme, C. and Miorner, H. (1989) Evaluation of Four Qualitative Methods for Detection of Beta Lactamase Production in Staphylococcus aureus and Micrococcus Species. European Journal of Clinical Microbiology and Infectious Diseases, 8, 962-967. http://dx.doi.org/10.1007/BF01967566
[23] Nitrocefin|CAS 41906-86-9|Calbiochem | 5MG. http://www.merckmillipore.com
[24] Montgomery, K., Raymundo, L. and Drew, W.L. (1979) Chromogenic Cephalosporin Spot Test to Detect Beta-Lactamase in Clinically Significant Bacteria. Journal of Clinical Microbiology, 9, 205-207.
[25] Plested, S.J., Simpson, I.N. and James, M. (1983) The Detection of Bacterial β Lactamase and Their Identification by Analytical Isoelectric Focusing. In: Russel, A.D. and Quesnel, L.B., Eds., Antibiotics: Assessment of Antimicrobial Activity and Resistance, Academic Press Inc., London.
[26] Kilic, E. and Cirak, Y. (2006) Comparison of Staphylococcal Beta Lactamase Detection Methods. Fabad Journal of Pharmaceutical Sciences, 31, 79-84.
[27] Devapriya, F., Ramesh, R., Khan, A.K.S. and Shanmugam, J. (2013) β-Lactamase Production of Staphylococcus Aureus: A Comparison Study of Different Iodometric Methods. Gulf Medical Journal, 2, 16-21.
[28] Sykes, R.B. and Matthew, M. (1976) The Beta-Lactamases of Gram-Negative Bacteria and Their Role in Resistance to Beta Lactam Antibiotics. Journal of Antimicrobial Chemotherapy, 2, 115-157. http://dx.doi.org/10.1093/jac/2.2.115
[29] Clinical and Laboratory Standards Institute (2006) Methods for Dilution Antimicrobial Susceptibility Test for Bacteria that Grow Aerobically. M7. Clinical and Laboratory Standards Institute, Wayne.

  
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