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. 1973 Jul;26(1):43–48. doi: 10.1128/am.26.1.43-48.1973

Preparation, Sensitivity, and Specificity of Limulus Lysate for Endotoxin Assay

James H Jorgensen 1,2, Rodney F Smith 1,2
PMCID: PMC379714  PMID: 4199336

Abstract

Limulus amoebocyte lysate was prepared from a total of 180 crabs during 1971 and 1972 by using a slightly modified lysate preparation procedure. Marked variability of lysate potency was noted both years. In addition, lysate quality appeared diminished in 1972 as compared with 1971. Different lysate batches were evaluated for potency by using a variety of endotoxin preparations. Variations in batch potencies were observed, but little variation in reactivity among different endotoxin preparations was noted. Use of potent lysate batches allowed detection of endotoxin concentrations as low as 100 pg/ml. No endotoxinlike activity was observed from 11 different strains of yeast by use of the Limulus assay.

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Selected References

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

  1. Caridis D. T., Reinhold R. B., Woodruff P. W., Fine J. Endotoxaemia in man. Lancet. 1972 Jun 24;1(7765):1381–1385. doi: 10.1016/s0140-6736(72)91108-7. [DOI] [PubMed] [Google Scholar]
  2. Cooper J. F., Hochstein H. D., Seligmann E. B., Jr The Limulus test for endotoxin (Pyrogen) in radiopharmaceuticals and biologicals. Bull Parenter Drug Assoc. 1972 Jul-Aug;26(4):153–162. [PubMed] [Google Scholar]
  3. Cooper J. F., Levin J., Wagner H. N., Jr Quantitative comparison of in vitro and in vivo methods for the detection of endotoxin. J Lab Clin Med. 1971 Jul;78(1):138–148. [PubMed] [Google Scholar]
  4. Cuevas P., Fine J. Role of intraintestinal endotoxin in death from peritonitis. Surg Gynecol Obstet. 1972 Jun;134(6):953–957. [PubMed] [Google Scholar]
  5. Cutler J. E., Friedman L., Milner K. C. Biological and chemical characterization of toxic substances from Candida albicans. Infect Immun. 1972 Oct;6(4):616–627. doi: 10.1128/iai.6.4.616-627.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. Reinhold R. B., Fine J. A technique for quantitative measurement of endotoxin in human plasma. Proc Soc Exp Biol Med. 1971 May;137(1):334–340. doi: 10.3181/00379727-137-35572. [DOI] [PubMed] [Google Scholar]
  9. Selzer G. B. Preparation of a purified lipopolysaccharide for pyrogen testing. Bull Parenter Drug Assoc. 1970 May-Jun;24(3):153–156. [PubMed] [Google Scholar]
  10. Ward P. A., Hill J. H. Detection of lipopolysaccharide (LPS): an improved method for isolation of the Limulus extract. Proc Soc Exp Biol Med. 1972 Dec;141(3):898–900. doi: 10.3181/00379727-141-36897. [DOI] [PubMed] [Google Scholar]
  11. Yin E. T., Galanos C., Kinsky S., Bradshaw R. A., Wessler S., Lüderitz O., Sarmiento M. E. Picogram-sensitive assay for endotoxin: gelation of Limulus polyphemus blood cell lysate induced by purified lipopolysaccharides and lipid A from Gram-negative bacteria. Biochim Biophys Acta. 1972 Jan 28;261(1):284–289. doi: 10.1016/0304-4165(72)90340-6. [DOI] [PubMed] [Google Scholar]
  12. Young N. S., Levin J., Prendergast R. A. An invertebrate coagulation system activated by endotoxin: evidence for enzymatic mediation. J Clin Invest. 1972 Jul;51(7):1790–1797. doi: 10.1172/JCI106980. [DOI] [PMC free article] [PubMed] [Google Scholar]

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