Abstract
A screening assay for phospholipase C using a chromogenic substrate incorporated into agar medium is described. The assay directly visualizes phospholipase C activity of mycoplasma lysates and membranes on agar plates, or the activity may be measured by spectrophotometry. The results from the assay confirm the presence in Ureaplasma urealyticum of phospholipase C, which is predominantly localized in the membrane fraction. The procedure has the potential to screen phospholipase C activity in other mycoplasmas and microorganisms in general.
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- Baine W. B., Rasheed J. K., Mackel D. C., Bopp C. A., Wells J. G., Kaufmann A. F. Exotoxin activity associated with the Legionnaires disease bacterium. J Clin Microbiol. 1979 Mar;9(3):453–456. doi: 10.1128/jcm.9.3.453-456.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bejar R., Curbelo V., Davis C., Gluck L. Premature labor. II. Bacterial sources of phospholipase. Obstet Gynecol. 1981 Apr;57(4):479–482. [PubMed] [Google Scholar]
- De Silva N. S., Quinn P. A. Endogenous activity of phospholipases A and C in Ureaplasma urealyticum. J Clin Microbiol. 1986 Feb;23(2):354–359. doi: 10.1128/jcm.23.2.354-359.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kurioka S., Matsuda M. Phospholipase C assay using p-nitrophenylphosphoryl-choline together with sorbitol and its application to studying the metal and detergent requirement of the enzyme. Anal Biochem. 1976 Sep;75(1):281–289. doi: 10.1016/0003-2697(76)90078-6. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Legakis N. J., Papavassiliou J. Thin-layer chromatographic technique for rapid detection of bacterial phospholipases. J Clin Microbiol. 1975 Nov;2(5):373–376. doi: 10.1128/jcm.2.5.373-376.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Owens J. J. The egg yolk reaction produced by several species of bacteria. J Appl Bacteriol. 1974 Mar;37(1):137–148. doi: 10.1111/j.1365-2672.1974.tb00424.x. [DOI] [PubMed] [Google Scholar]
- Wiley C. A., Quinn P. A. Enzyme-linked immunosorbent assay for detection of specific antibodies to Ureaplasma urealyticum serotypes. J Clin Microbiol. 1984 Mar;19(3):421–426. doi: 10.1128/jcm.19.3.421-426.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]