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. 1981 Jun;41(6):1316–1320. doi: 10.1128/aem.41.6.1316-1320.1981

Automation of the Limulus amoebocyte lysate test by using the Abbott MS-2 microbiology system.

J H Jorgensen, G A Alexander
PMCID: PMC243917  PMID: 7247396

Abstract

A rapid, automated method for the performance of the Limulus amoebocyte lysate endotoxin assay has been developed by using the Abbott MS-2 Microbiology System. This instrument automatically determines sequential changes in the optical density of up to 176 samples at 1- or 5-min increments during a 1-h assay period. Graphic representation of optical density changes can be viewed on a cathode-ray tube or reproduced by using a hard-copy printer. Limulus amoebocyte lysate preparations that were obtained from different commercial producers and that had similar endotoxin sensitivities by the conventional gelation method varied somewhat in reactivity when determinations were based upon rate changes in optical density. Lysates from Associates of Cape Cod, Difco Laboratories, and M. A. Bioproducts were the most readily adaptable to the MS-2 System. Use of the MS-2 system increased the sensitivity of these preparations from 60- to 250-fold, and as little as 1 pg/ml was detected. Adaptation of the MS-2 instrument for this purpose provides an objective, reproducible, automated method for the performance of Limulus amoebocyte lysate tests on a variety of fluids.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Elin R. J., Robinson R. A., Levine A. S., Wolff S. M. Lack of clinical usefulness of the limulus test in the diagnosis of endotoxemia. N Engl J Med. 1975 Sep 11;293(11):521–524. doi: 10.1056/NEJM197509112931102. [DOI] [PubMed] [Google Scholar]
  2. Evans T. M., Schillinger J. E., Stuart D. G. Rapid determination of bacteriological water quality by using limulus lysate. Appl Environ Microbiol. 1978 Feb;35(2):376–382. doi: 10.1128/aem.35.2.376-382.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Frauch P. Letter: Slide test as a micromethod of a modified Limulus endotoxin test. J Pharm Sci. 1974 May;63(5):808–809. doi: 10.1002/jps.2600630542. [DOI] [PubMed] [Google Scholar]
  4. Harrison S. J., Tsuji K., Enzinger R. M. Application of LAL for detection of endotoxin in antibiotic preparations. Prog Clin Biol Res. 1979;29:353–365. [PubMed] [Google Scholar]
  5. Jorgensen J. H., Jones P. M. Comparative evaluation of the Limulus assay and the direct Gram stain for detection of significant bacteriuria. Am J Clin Pathol. 1975 Jan;63(1):142–148. doi: 10.1093/ajcp/63.3.142. [DOI] [PubMed] [Google Scholar]
  6. Jorgensen J. H., Lee J. C., Alexander G. A., Wolf H. W. Comparison of Limulus assay, standard plate count, and total coliform count for microbiological assessment of renovated wastewater. Appl Environ Microbiol. 1979 May;37(5):928–931. doi: 10.1128/aem.37.5.928-931.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jorgensen J. H., Lee J. C. Rapid diagnosis of gram-negative bacterial meningitis by the Limulus endotoxin assay. J Clin Microbiol. 1978 Jan;7(1):12–17. doi: 10.1128/jcm.7.1.12-17.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jorgensen J. H., Smith R. F. Preparation, sensitivity, and specificity of Limulus lysate for endotoxin assay. Appl Microbiol. 1973 Jul;26(1):43–48. doi: 10.1128/am.26.1.43-48.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Levin J., Bang F. B. Clottable protein in Limulus; its localization and kinetics of its coagulation by endotoxin. Thromb Diath Haemorrh. 1968 Mar 31;19(1):186–197. [PubMed] [Google Scholar]
  10. Levin J., Poore T. E., Young N. S., Margolis S., Zauber N. P., Townes A. S., Bell W. R. Gram-negative sepsis: detection of endotoxemia with the limulus test. With studies of associated changes in blood coagulation, serum lipids, and complement. Ann Intern Med. 1972 Jan;76(1):1–7. doi: 10.7326/0003-4819-76-1-1. [DOI] [PubMed] [Google Scholar]
  11. McBeath J., Forster R. K., Rebell G. Diagnostic limulus lysate assay for endophthalmitis and keratitis. Arch Ophthalmol. 1978 Jul;96(7):1265–1267. doi: 10.1001/archopht.1978.03910060091020. [DOI] [PubMed] [Google Scholar]
  12. McCracken A. W., Martin W. J., McCarthy L. R., Schwab D. A., Cooper B. H., Helgeson N. G., Prowant S., Robson J. Evaluation of the MS-2 system for rapid identification of Enterobacteriaceae. J Clin Microbiol. 1980 Nov;12(5):684–689. doi: 10.1128/jcm.12.5.684-689.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Munford R. S. Quantitative Limulus lysate assay for endotoxin activity: aggregation of radioiodinated coagulogen monomers. Anal Biochem. 1978 Dec;91(2):509–515. doi: 10.1016/0003-2697(78)90537-7. [DOI] [PubMed] [Google Scholar]
  14. Nachum R., Lipsey A., Siegel S. E. Rapid detection of gram-negative bacterial meningitis by the limulus lysate test. N Engl J Med. 1973 Nov 1;289(18):931–934. doi: 10.1056/NEJM197311012891801. [DOI] [PubMed] [Google Scholar]
  15. Prior R. B., Spagna V. A. Adaptation of a microdilution procedure to the Limulus lysate assay for endotoxin. J Clin Microbiol. 1979 Sep;10(3):394–395. doi: 10.1128/jcm.10.3.394-395.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ross S., Rodriguez W., Controni G., Korengold G., Watson S., Khan W. Limulus lysate test for gram-negative bacterial meningitis. Bedside application. JAMA. 1975 Sep 29;233(13):1366–1369. [PubMed] [Google Scholar]
  17. Spagna V. A., Prior R. B., Perkins R. L. Rapid presumptive diagnosis of gonococcal cervicitis by the limulus lysate assay. Am J Obstet Gynecol. 1980 Jul 1;137(5):595–599. doi: 10.1016/0002-9378(80)90702-4. [DOI] [PubMed] [Google Scholar]
  18. Spagna V. A., Prior R. B., Perkins R. L. Rapid presumptive diagnosis of gonococcal urethritis in men by the limulus lysate test. Br J Vener Dis. 1979 Jun;55(3):179–182. doi: 10.1136/sti.55.3.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Teller J. D., Kelly K. M. A turbidimetric Limulus amebocyte assay for the quantitative determination of Gram negative bacterial endotoxin. Prog Clin Biol Res. 1979;29:423–433. [PubMed] [Google Scholar]
  20. Thornsberry C., Anhalt J. P., Washington J. A., 2nd, McCarthy L. R., Schoenknecht F. D., Sherris J. C., Spencer H. J. Clinical laboratory evaluation of the Abbott MS-2 automated antimicrobial susceptibility testing system: report of a collaborative study. J Clin Microbiol. 1980 Sep;12(3):375–390. doi: 10.1128/jcm.12.3.375-390.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wachtel R. E., Tsuji K. Comparison of limulus amebocyte lysates and correlation with the United States Pharmacopeial pyrogen test. Appl Environ Microbiol. 1977 Jun;33(6):1265–1269. doi: 10.1128/aem.33.6.1265-1269.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wolters R. W., Jorgensen J. H., Calzada E., Poirier R. H. Limulus lysate assay for early detection of certain Gram-negative corneal infections. Arch Ophthalmol. 1979 May;97(5):875–877. doi: 10.1001/archopht.1979.01020010433006. [DOI] [PubMed] [Google Scholar]

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