1887

Abstract

A polyclonal antibody raised to spores reacted strongly and specifically with the immunizing strain and to a number of related species of , as determined by dot immunoblotting. An indirect immunomagnetic capture method was developed for the recovery of the target organism from sterile and non-sterile soil, using sheep anti-rabbit M-280 Dynabeads. The effects of different soil blocking agents, antibody labelling concentrations and spore/Dynabead capture times on the recovery of spores were investigated. Pre-blocking of antibody binding sites within the soil, with either 2% partially hydrolysed gelatin or 10% skimmed milk, was essential prior to immunomagnetic capture. Increasing the capture time from 15 to 60 min did not affect spore recovery; however, a 10-fold decline in the magnetic bead concentration did result in a significantly lower recovery of spores from soil. was selectively enriched (1:190-fold) when present as a mixed population with in sterile soil. The indirect immunomagnetic capture method was used to selectively recover S. fragile spores seeded into non-sterile soil, although some background binding of non-target bacteria was noted. The target was successfully recovered from a sterile soil microcosm after 14 d incubation and the capture rate was increased by the inclusion of an initial soil dispersion and biomass concentration procedure, using the ion-exchange resin Chelex 100.

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-141-9-2149
1995-09-01
2022-01-26
Loading full text...

Full text loading...

/deliver/fulltext/micro/141/9/mic-141-9-2149.html?itemId=/content/journal/micro/10.1099/13500872-141-9-2149&mimeType=html&fmt=ahah

References

  1. Bifulco J.M., Schaefer F.W. 1993; Antibody-magnetite method for the selective concentration of Giardia lamblia cysts from water samples.. Appl Environ Microbiol 59:772–776
    [Google Scholar]
  2. Blackburn de C.W., Patel P.D., Gibbs P.A. 1991; Separation and detection of Salmonellae using immunomagnetic particles.. Biofouling 5:143–156
    [Google Scholar]
  3. Bohool B.B., Schmidt E.L. 1980; The immunofluorescence approach in microbial ecology.. Adv Microb Ecol 4:203–241
    [Google Scholar]
  4. Bradford M.M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.. Anal Biochem 72:248–254
    [Google Scholar]
  5. Christensen B., Torsvik T., Lien T. 1992; Immunomagnetically captured thermophilic sulphate-reducing bacteria from North Sea oil field waters.. Appl Environ Microbiol 58:1244–1248
    [Google Scholar]
  6. Conway de Macario E., Wolin M.J., Macario A.J.L. 1982; Specific antisera and immunological procedures for characterization of methanogenic bacteria.. J Bacteriol 149:320–328
    [Google Scholar]
  7. Ford S.F., Olsen B. 1988; Methods for detecting genetically engineered microorganisms in the environment.. Adv Microb Ecol 10:45–79
    [Google Scholar]
  8. Fratamico P.M., Schultz F.J., Buchanan R.L. 1992; Rapid isolation of Escherichia coli O157 :H7 from enrichment cultures of foods using an immunomagnetic separation method.. Food Microbiol 9:105–113
    [Google Scholar]
  9. Gaudernack G., Ugelstad T., Thoresby E. 1986; Isolation of pure functionally active CD8+ T cells. Positive selection with monoclonal antibodies directly conjugated to monosized magnetic microspheres.. J Immunol Meth 90:179–187
    [Google Scholar]
  10. Gordon J., Billing P. 1988; Dot immunobinding, general principles and procedures.. In CRC Handbook of Immunoblotting of Proteins 1 section 2, pp. 27–30 Bjerrum O.J., Heegaard N.H.H. Edited by Boca Raton, FL: CRC Press;
    [Google Scholar]
  11. Gruhn B.R., Haefer A., Mueller W., Andrae H., Danan &Zintl F. 1991; Model experiments for immunomagnetic elimination of leukemic cells from human bone marrow: presentation of a novel magnetic separation system.. Immunobiology 183:374–385
    [Google Scholar]
  12. Harboe N.M.G., Ingild A. 1983; Immunization, isolation of immunoglobulins and antibody titre determination.. Scand J Immunol 17:345–351
    [Google Scholar]
  13. Hayakawa M., Nonomura H. 1989; A new method for the intensive isolation of actinomycetes from soil.. Actinomycetologica 3:95–104
    [Google Scholar]
  14. Herron P.R., Wellington E.M.H. 1990; New method for extraction of streptomycete spores from soil and application to the study of lysogeny in sterile amended and nonsterile soil.. Appl Environ Microbiol 56:1406–1412
    [Google Scholar]
  15. Hopwood D.A., Bibb M.J., Chafer K.F., Kieser T., Bruton C.J., Kieser M., Lydiate D.J., Smith C.P., Ward J.M., Schrempf H. 1985 In Genetic Manipulation of Streptomyces: a Eaboratory Manual pp. 3–5 Norwich: John Innes Foundation;
    [Google Scholar]
  16. Johne B., Jarp J., Haaheim L.R. 1992; Staphylococcus aureus exopolysaccharide in vivo demonstrated by immunomagnetic separation and electron microscopy.. J Clin Microbiol 27:1631–1635
    [Google Scholar]
  17. Luk J.M.C., Lindberg A.A. 1991; Rapid and sensitive detection of Salmonella (O:67) by immunomagnetic monoclonal antibody- based assays.. J Immunol Meth 137:1–8
    [Google Scholar]
  18. Lund A., Helleman A.L., Vartdal F. 1988; Rapid isolation of K88+ Escherichia coli by using immunomagnetic particles.. J Clin Microbiol US2572–2575
    [Google Scholar]
  19. Mansfield L.P., Forsythe S.J. 1993; Immunomagnetic separation as an alternative to enrichment broths for Salmonella detection.. Lett Appl Microbiol 16:122–125
    [Google Scholar]
  20. Mason J., Burns R.G. 1990; Production of a monoclonal antibody specific for a Flavobacterium species isolated from soil.. FEMS Microbiol Ecol 73:299–308
    [Google Scholar]
  21. Morgan J.A.W., Winstanley C., Pickup R.W., Jones J.G., Saunders J.R. 1989; Direct phenotypic and genotypic detection of a recombinant pseudomonad population released into lake water.. Appl Environ Microbiol 55:2537–2544
    [Google Scholar]
  22. Morgan J.A.W., Winstanley C., Pickup R.W., Saunders J.R. 1991; Rapid immunomagnetic capture of Pseudomonas putida cells from lake water by using bacterial flagella.. Appl Environ Microbiol 57:503–509
    [Google Scholar]
  23. Nelson D., Bathgate A.J., Poxton I.R. 1991; Monoclonal antibodies as probes for detecting lipopolysaccharide expression on Escherichia coli from different growth conditions.. J Gen Microbiol 137:2741–2751
    [Google Scholar]
  24. Nolan R.D., Cross T. 1988; Isolation and screening of actinomycetes.. In Actinomycetes in Biotechnology pp. 1–32 Goodfellow M., Williams S.T., Mordarski M. Edited by London: Academic Press;
    [Google Scholar]
  25. Nonomura H. 1989; Genus Streptosporangium Couch 1955.. In Bergefs Manual of Systematic Bacteriology 4: pp. 2545–2551 Williams S.T., Sharpe M., Holt J.G. Edited by Baltimore: Williams & Wilkins;
    [Google Scholar]
  26. Parmer N., Easter M.C., Forsythe S.J. 1992; The detection of Salmonella enteritidis and T.typhimurium using immunomagnetic separation and conductance microbiology.. Lett Appl Microbiol 15:175–178
    [Google Scholar]
  27. Petersen R.G. 1985; Separation of means.. In Design and Analysis of Experiments (Statistics: Textbooks and Monographs) 66 pp. 72–111 New York: Marcel-Dekker;
    [Google Scholar]
  28. Pickup R.W. 1991; Development of molecular methods for the detection of specific bacteria in the environment.. J Gen Microbiol 137:1009–1019
    [Google Scholar]
  29. Postma J., van Elsas J.D., Govaert J.M, Veen J.A. 1988; Thedynamics of Rhizpbium leguminosarum biovar trifolii introduced into soil as determined by immunofluorescence and selective plating techniques.. FEMS Microbiol Ecol 53:251–260
    [Google Scholar]
  30. Schmidt E.L. 1974; Quantitative autecological study of micro-organisms in soil by immunofluorecsence.. Soil Sci 118:141–149
    [Google Scholar]
  31. Shearer M.C. 1987; Methods for the isolation of non- streptomycete actinomycetes. Developments in industrial microbiology.. J Ind Microbiol 28 Suppl. 2 91–97 Pierce G. Edited by
    [Google Scholar]
  32. Silvestri F., Banavali S., Yin M., Gopal V., Savignano C., Baccarani M., Preisler H.D. 1992; CD34-positive cell selection by immunomagnetic beads and chymopapain.. Haematologica 77:307–310
    [Google Scholar]
  33. Skjerve E., Olsvik O. 1991; Immunomagnetic separation of Salmonella from foods.. Int J Food Microbiol 14:11–18
    [Google Scholar]
  34. Skjerve E., Rorvik L.M., Olsvik O. 1990; Detection of Listeria monocytogenes in foods by immunomagnetic separation.. Appl Environ Microbiol 56:3478–3481
    [Google Scholar]
  35. Sokal R.R., Rohlf F.J. 1969; Assumptions of ANOV.. In Biometry: Principles and Practice of Statistics in Biological Research pp. 235–246 San Francisco: W.H. Freeman;
    [Google Scholar]
  36. Tomoyasu T. 1992; Development of the immunomagnetic enrichment method selective for Vibrio parahaemolyticus serotype K and its application to a food poisoning study.. Appl Environ Microbiol 58:2679–2682
    [Google Scholar]
  37. Trevors J.T., Van Elsas J.D., Lee H., Wolters A.C. 1993; Survival of alginate-encapsulated Pseudomonas fluorescens cells in soil.. Appl Microbiol Biotechnol 39:637–643
    [Google Scholar]
  38. Turpin P.E., Maycroft K.A., Rowlands C.L., Wellington E.M.H. 1993a; Viable but non-culturable salmonellas in soil.. J Appl Bacteriol 74:421–427
    [Google Scholar]
  39. Turpin P.E., Maycroft K.A., Rowlands C.L., Wellington E.M.H. 1993b; An ion-exchange based extraction method for the detection of salmonellas in soil.. J Appl Bacteriol 74:181–190
    [Google Scholar]
  40. Vermunt A.E.M., Franken A.A.J.M., Beumer R.R. 1992; Isolation of salmonellas by immunomagnetic separation.. J Appl Bacteriol 72:112–118
    [Google Scholar]
  41. Wellington E.M.H., Toth I.K. 1994; Actinomycetes.. In Methods of Soil Analysis: part 2, Microbiological and Biochemical Properties of Soils, 3rd edn. pp. 269–290 Weaver R.W., Angle J.S., Bottomly P.S. Edited by Madison, WI: Soil Science Society of America;
    [Google Scholar]
  42. Wellington E.M.H., Cresswell N.C., Saunders V.A. 1990; Growth and survival of streptomycete inoculants and extent of plasmid transfer in sterile and nonsterile soil.. Appl Environ Microbiol 56:1413–1419
    [Google Scholar]
  43. Wipat A., Wellington E.M.H., Saunders V.A. 1994; Immuno-logical detection and recovery of genetically manipulated streptomycetes from soil.. Microbiology 140:2067–2076
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-141-9-2149
Loading
/content/journal/micro/10.1099/13500872-141-9-2149
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error