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. 1989 Jan;27(1):182–184. doi: 10.1128/jcm.27.1.182-184.1989

Detection of sialidase (neuraminidase) activity in Actinomyces species by using 2'-(4-methylumbelliferyl)alpha-D-N-acetylneuraminic acid in a filter paper spot test.

B J Moncla 1, P Braham 1
PMCID: PMC267258  PMID: 2643620

Abstract

A rapid method for the detection of acetylneuraminyl hydrolase, EC 3.2.1.18 (sialidase or neuraminidase), was developed by using 2'-(4-methylumbelliferyl)alpha-D-N-acetylneuraminic acid as substrate in a filter paper spot test. The method was compared to conventional assays that use 2'-(4-methylumbelliferyl)alpha-D-N-acetylneuraminic acid and bovine submaxillary mucin and was found to be in excellent agreement. Organisms with greater than 10 U of enzyme activity (in nanomoles per minute per milligram of cell protein) gave positive reactions, while those with 2.7 to 9.0 U gave only weak reactions. Isolates with less than 2.7 U of activity were detected upon prolonged incubation. Sialidase activity was detected in 79% of 71 clinical isolates representing five species of Actinomyces. The percentage of sialidase-producing isolates of each species varied considerably: Actinomyces israelii, 63%; A. meyeri, 73%; A. naeslundii, 85%; A. odontolyticus, 73%; and A. viscosus, 100%.

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Selected References

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  1. Brown J. G., Straus D. C. Characterization of neuraminidases produced by various serotypes of group B streptococci. Infect Immun. 1987 Jan;55(1):1–6. doi: 10.1128/iai.55.1.1-6.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Costello A. H., Cisar J. O., Kolenbrander P. E., Gabriel O. Neuraminidase-dependent hamagglutination of human erythrocytes by human strains of Actinomyces viscosus and Actinomyces naeslundii. Infect Immun. 1979 Nov;26(2):563–572. doi: 10.1128/iai.26.2.563-572.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Drzeniek R. Viral and bacterial neuraminidases. Curr Top Microbiol Immunol. 1972;59:35–74. doi: 10.1007/978-3-642-65444-2_2. [DOI] [PubMed] [Google Scholar]
  4. Hill G. B., Ayers O. M., Kohan A. P. Characteristics and sites of infection of Eubacterium nodatum, Eubacterium timidum, Eubacterium brachy, and other asaccharolytic eubacteria. J Clin Microbiol. 1987 Aug;25(8):1540–1545. doi: 10.1128/jcm.25.8.1540-1545.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Jones A. H., Lee C. C., Moncla B. J., Robinovitch M. R., Birdsell D. C. Surface localization of sialic acid on Actinomyces viscosus. J Gen Microbiol. 1986 Dec;132(12):3381–3391. doi: 10.1099/00221287-132-12-3381. [DOI] [PubMed] [Google Scholar]
  6. Kelly R. T., Farmer S., Greiff D. Neuraminidase activities of clinical isolates of Diplococcus pneumoniae. J Bacteriol. 1967 Jul;94(1):272–273. doi: 10.1128/jb.94.1.272-273.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Leprat R., Michel-Briand Y. Extracellular neuraminidase production by a strain of Pseudomonas aeruginosa isolated from cystic fibrosis. Ann Microbiol (Paris) 1980 Nov-Dec;131B(3):209–222. [PubMed] [Google Scholar]
  9. Müller H. E. Untersuchungen in vitro über di Neuraminidase der Pasteurella multocida. Zentralbl Bakteriol Orig A. 1971 Jul;217(3):326–344. [PubMed] [Google Scholar]
  10. O'Brien M., Colwell R. R. A rapid test for chitinase activity that uses 4-methylumbelliferyl-N-acetyl-beta-D-glucosaminide. Appl Environ Microbiol. 1987 Jul;53(7):1718–1720. doi: 10.1128/aem.53.7.1718-1720.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. O'Toole R. D., Goode L., Howe C. Neuraminidase activity in bacterial meningitis. J Clin Invest. 1971 May;50(5):979–985. doi: 10.1172/JCI106591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Palese P., Bucher D., Kilbourne E. D. Applications of a synthetic neuraminidase substrate. Appl Microbiol. 1973 Feb;25(2):195–201. doi: 10.1128/am.25.2.195-201.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Pardoe G. I. The inducible neuraminidase (N-acyl-neuraminyl hydrolase EC 3.2.1.18) of Klebsiella aerogenes NCIB 9479. Pathol Microbiol (Basel) 1970;35(6):361–376. doi: 10.1159/000162248. [DOI] [PubMed] [Google Scholar]
  14. Potier M., Mameli L., Bélisle M., Dallaire L., Melançon S. B. Fluorometric assay of neuraminidase with a sodium (4-methylumbelliferyl-alpha-D-N-acetylneuraminate) substrate. Anal Biochem. 1979 Apr 15;94(2):287–296. doi: 10.1016/0003-2697(79)90362-2. [DOI] [PubMed] [Google Scholar]
  15. Schauer R. Chemistry, metabolism, and biological functions of sialic acids. Adv Carbohydr Chem Biochem. 1982;40:131–234. doi: 10.1016/s0065-2318(08)60109-2. [DOI] [PubMed] [Google Scholar]
  16. Skoza L., Mohos S. Stable thiobarbituric acid chromophore with dimethyl sulphoxide. Application to sialic acid assay in analytical de-O-acetylation. Biochem J. 1976 Dec 1;159(3):457–462. doi: 10.1042/bj1590457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Tallman J. F., Brady R. O. The purification and properties of a mammalian neuraminidase (sialidase). Biochim Biophys Acta. 1973 Feb 15;293(2):434–443. doi: 10.1016/0005-2744(73)90350-1. [DOI] [PubMed] [Google Scholar]
  18. Tallman J. F., Fishman P. H., Henneberry R. C. Determination of sialidase activities in HeLa cells using gangliosides specifically labeled in N-acetylneuraminic acid. Arch Biochem Biophys. 1977 Aug;182(2):556–562. doi: 10.1016/0003-9861(77)90537-9. [DOI] [PubMed] [Google Scholar]
  19. Tuppy H., Palese P. A chromogenic substrate for the investigation of neuraminidases. FEBS Lett. 1969 Apr;3(1):72–75. doi: 10.1016/0014-5793(69)80100-6. [DOI] [PubMed] [Google Scholar]

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