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. 1995 Sep;33(9):2498–2500. doi: 10.1128/jcm.33.9.2498-2500.1995

Immunological detection of penicillin-binding protein 2' of methicillin-resistant staphylococci by using monoclonal antibodies prepared from synthetic peptides.

M Saito 1, K Sekiguchi 1, R Yajima 1, M Hina 1, R C Doss 1, H Kanno 1
PMCID: PMC228456  PMID: 7494059

Abstract

Two synthetic peptides 31 and 32 amino acids in length were prepared as deduced from a known amino acid sequence of penicillin-binding protein 2' (PBP2') of methicillin-resistant Staphylococcus aureus. Two monoclonal antibodies were generated from fused cells of myeloma cells and splenic cells of mice immunized with the synthetic peptides. Western blot (immunoblot) analysis demonstrated specific binding of the antibodies to PBP2' of a methicillin-resistant S. aureus strain. An immunoradiometric assay was developed by using these antibodies for simple detection of PBP2'.

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Selected References

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  1. Bolton A. E., Hunter W. M. The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent. Biochem J. 1973 Jul;133(3):529–539. doi: 10.1042/bj1330529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Gerberding J. L., Miick C., Liu H. H., Chambers H. F. Comparison of conventional susceptibility tests with direct detection of penicillin-binding protein 2a in borderline oxacillin-resistant strains of Staphylococcus aureus. Antimicrob Agents Chemother. 1991 Dec;35(12):2574–2579. doi: 10.1128/aac.35.12.2574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Kanda K., Suzuki E., Hiramatsu K., Oguri T., Miura H., Ezaki T., Yokota T. Identification of a methicillin-resistant strain of Staphylococcus caprae from a human clinical specimen. Antimicrob Agents Chemother. 1991 Jan;35(1):174–176. doi: 10.1128/aac.35.1.174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  5. Ligozzi M., Rossolini G. M., Tonin E. A., Fontana R. Nonradioactive DNA probe for detection of gene for methicillin resistance in Staphylococcus aureus. Antimicrob Agents Chemother. 1991 Mar;35(3):575–578. doi: 10.1128/aac.35.3.575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Murakami K., Minamide W., Wada K., Nakamura E., Teraoka H., Watanabe S. Identification of methicillin-resistant strains of staphylococci by polymerase chain reaction. J Clin Microbiol. 1991 Oct;29(10):2240–2244. doi: 10.1128/jcm.29.10.2240-2244.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Pierre J., Williamson R., Bornet M., Gutmann L. Presence of an additional penicillin-binding protein in methicillin-resistant Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus hominis, and Staphylococcus simulans with a low affinity for methicillin, cephalothin, and cefamandole. Antimicrob Agents Chemother. 1990 Sep;34(9):1691–1694. doi: 10.1128/aac.34.9.1691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ryffel C., Tesch W., Birch-Machin I., Reynolds P. E., Barberis-Maino L., Kayser F. H., Berger-Bächi B. Sequence comparison of mecA genes isolated from methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis. Gene. 1990 Sep 28;94(1):137–138. doi: 10.1016/0378-1119(90)90481-6. [DOI] [PubMed] [Google Scholar]
  9. Skulnick M., Simor A. E., Gregson D., Patel M., Small G. W., Kreiswirth B., Hathoway D., Low D. E. Evaluation of commercial and standard methodology for determination of oxacillin susceptibility in Staphylococcus aureus. J Clin Microbiol. 1992 Aug;30(8):1985–1988. doi: 10.1128/jcm.30.8.1985-1988.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Song M. D., Wachi M., Doi M., Ishino F., Matsuhashi M. Evolution of an inducible penicillin-target protein in methicillin-resistant Staphylococcus aureus by gene fusion. FEBS Lett. 1987 Aug 31;221(1):167–171. doi: 10.1016/0014-5793(87)80373-3. [DOI] [PubMed] [Google Scholar]
  11. Tesch W., Strässle A., Berger-Bächi B., O'Hara D., Reynolds P., Kayser F. H. Cloning and expression of methicillin resistance from Staphylococcus epidermidis in Staphylococcus carnosus. Antimicrob Agents Chemother. 1988 Oct;32(10):1494–1499. doi: 10.1128/aac.32.10.1494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Tokue Y., Shoji S., Satoh K., Watanabe A., Motomiya M. Comparison of a polymerase chain reaction assay and a conventional microbiologic method for detection of methicillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother. 1992 Jan;36(1):6–9. doi: 10.1128/aac.36.1.6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Townsend D. E., Ashdown N., Bolton S., Bradley J., Duckworth G., Moorhouse E. C., Grubb W. B. The international spread of methicillin-resistant Staphylococcus aureus. J Hosp Infect. 1987 Jan;9(1):60–71. doi: 10.1016/0195-6701(87)90097-1. [DOI] [PubMed] [Google Scholar]
  14. Ubukata K., Nakagami S., Nitta A., Yamane A., Kawakami S., Sugiura M., Konno M. Rapid detection of the mecA gene in methicillin-resistant staphylococci by enzymatic detection of polymerase chain reaction products. J Clin Microbiol. 1992 Jul;30(7):1728–1733. doi: 10.1128/jcm.30.7.1728-1733.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ubukata K., Nonoguchi R., Song M. D., Matsuhashi M., Konno M. Homology of mecA gene in methicillin-resistant Staphylococcus haemolyticus and Staphylococcus simulans to that of Staphylococcus aureus. Antimicrob Agents Chemother. 1990 Jan;34(1):170–172. doi: 10.1128/aac.34.1.170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ubukata K., Yamashita N., Konno M. Occurrence of a beta-lactam-inducible penicillin-binding protein in methicillin-resistant staphylococci. Antimicrob Agents Chemother. 1985 May;27(5):851–857. doi: 10.1128/aac.27.5.851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Unal S., Hoskins J., Flokowitsch J. E., Wu C. Y., Preston D. A., Skatrud P. L. Detection of methicillin-resistant staphylococci by using the polymerase chain reaction. J Clin Microbiol. 1992 Jul;30(7):1685–1691. doi: 10.1128/jcm.30.7.1685-1691.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Utsui Y., Yokota T. Role of an altered penicillin-binding protein in methicillin- and cephem-resistant Staphylococcus aureus. Antimicrob Agents Chemother. 1985 Sep;28(3):397–403. doi: 10.1128/aac.28.3.397. [DOI] [PMC free article] [PubMed] [Google Scholar]

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