limulus test

Summary

Summary: Sensitive method for detection of bacterial endotoxins and endotoxin-like substances that depends on the in vitro gelation of Limulus amebocyte lysate (LAL), prepared from the circulating blood (amebocytes) of the horseshoe crab, by the endotoxin or related compound. Used for detection of endotoxin in body fluids and parenteral pharmaceuticals.

Top Publications

  1. Moesby L, Jensen S, Hansen E, Christensen J. A comparative study of Mono Mac 6 cells, isolated mononuclear cells and Limulus amoebocyte lysate assay in pyrogen testing. Int J Pharm. 1999;191:141-9 pubmed
    ..In conclusion, the MM6 assay is a good supplement to the current pyrogen assays for detection of LPS, S. aureus and S. typhimurium, but the MM6 assay could not detect A. niger, C. albicans and Influenza virus. ..
  2. Morris H, Monaco L, Steele A, Wainwright N. Setting a standard: the limulus amebocyte lysate assay and the assessment of microbial contamination on spacecraft surfaces. Astrobiology. 2010;10:845-52 pubmed publisher
  3. Gutsmann T, Howe J, Zahringer U, Garidel P, Schromm A, Koch M, et al. Structural prerequisites for endotoxic activity in the Limulus test as compared to cytokine production in mononuclear cells. Innate Immun. 2010;16:39-47 pubmed publisher
    ..These data are important since a reliable determination of endotoxin concentrations, in particular with respect to its ability to elicit severe infections, is of high interest. ..
  4. Hurley J. Endotoxemia: methods of detection and clinical correlates. Clin Microbiol Rev. 1995;8:268-92 pubmed
    ..However, with therapies designed to reduce levels of endotoxin, or to antagonize its effects, it is unclear whether clinical improvement occurs as a consequence of changes in the levels of endotoxemia...
  5. Hausmann M, Yulzari R, Lewis E, Saisky Y, Douvdevani A. Gel clot LAL assay in the initial management of peritoneal dialysis patients with peritonitis: a retrospective study. Nephrol Dial Transplant. 2000;15:680-3 pubmed
    ..Routine Limulus amoebocyte lysate assay of dialysate effluent from continuous ambulatory peritoneal dialysis patients presenting with peritonitis is recommended. ..
  6. Anderson W, Slawson R, Mayfield C. A review of drinking-water-associated endotoxin, including potential routes of human exposure. Can J Microbiol. 2002;48:567-87 pubmed
    ..It would appear that endotoxin originating in tap water can be inhaled but at present there is insufficient information available to quantify potential health risks. ..
  7. Morris A, Hillenbrand M, Finkelman M, George M, Singh V, Kessinger C, et al. Serum (1?3)-?-D-glucan levels in HIV-infected individuals are associated with immunosuppression, inflammation, and cardiopulmonary function. J Acquir Immune Defic Syndr. 2012;61:462-8 pubmed publisher
    ..These results implicate a new class of pathogen in HIV-associated microbial translocation and suggest a role in HIV progression and comorbidities. ..
  8. Matsumoto N, Takahashi G, Kojika M, Suzuki Y, Inoue Y, Inada K, et al. Interleukin-8 induces an elevation in the endotoxin activity assay (EAA) level: does the EAA truly measure the endotoxin level?. J Infect Chemother. 2013;19:825-32 pubmed publisher
    ..Overall, the EAA kit could not measure clinically relevant doses of endotoxin. Because IL-8 induced an increase in EAA level, it is suggested that the EAA level reflects the primed state of polymorphonuclear leukocytes. ..
  9. Ito H, Hirata M, Koga T. Hen egg-white lysozyme inhibits biological activities of lipopolysaccharides from periodontopathic bacteria. J Periodontal Res. 1997;32:295-9 pubmed
    ..The anti-endotoxic activity of HEL may be worthy of being intended for periodontal therapy. ..

More Information

Publications62

  1. Foto M, Plett J, Berghout J, Miller J. Modification of the Limulus amebocyte lysate assay for the analysis of glucan in indoor environments. Anal Bioanal Chem. 2004;379:156-62 pubmed
    ..Glucan content was proportional to spore surface area; however, some species contain higher relative spore glucan contents. ..
  2. Sakai H, Hisamoto S, Fukutomi I, Sou K, Takeoka S, Tsuchida E. Detection of lipopolysaccharide in hemoglobin-vesicles by Limulus amebocyte lysate test with kinetic-turbidimetric gel clotting analysis and pretreatment of surfactant. J Pharm Sci. 2004;93:310-21 pubmed
    ..0025 EU/mL. Pretreatment with C(12)E(10) would be applicable not only to HbVs but also to other drug delivery systems using phospholipid vesicles encapsulating or incorporating functional molecules. ..
  3. Dick H, Augustin A, Pakula T, Pfeiffer N. Endotoxins in ophthalmic viscosurgical devices. Eur J Ophthalmol. 2003;13:176-84 pubmed
    ..To measure the endotoxin concentration (EC) of 25 commercially available, hyaluronic acid- and hydroxypropylmethylcellulose-based (HPMC) ophthalmic viscosurgical devices (OVDs)...
  4. Uchiya K, Sugihara H. Biological and chemical studies on soluble protective antigen (SPA) from culture supernatant fluids of Salmonella enteritidis. Kansenshogaku Zasshi. 1990;64:802-8 pubmed
    ..In addition, SPA enhanced the clotting activity of Limulus amebocyte lysate. These results show that SPA possesses the properties of lipopolysaccharides isolated from strain 2547 by chemical procedures. ..
  5. Bjerre A, Brusletto B, Rosenqvist E, Namork E, Kierulf P, Øvstebø R, et al. Cellular activating properties and morphology of membrane-bound and purified meningococcal lipopolysaccharide. J Endotoxin Res. 2000;6:437-45 pubmed
    ..However, LAL activity was significantly higher for nOMV than for purified Nm-LPS and dOMV. The cellular responses of purified Nm-LPS and nOMV were reduced (>95%) by a specific anti-CD14-antibody. ..
  6. Rhee M, Kang D. Rapid and simple estimation of microbiological quality of raw milk using chromogenic Limulus amoebocyte lysate endpoint assay. J Food Prot. 2002;65:1447-51 pubmed
    ..This method may be successfully implemented to rapidly determine highly microbial contaminated raw milk (> 3.0 log10 CFU/ml of SPC). ..
  7. Hynes S, Ferris J, Szponar B, Wadstrom T, Fox J, O Rourke J, et al. Comparative chemical and biological characterization of the lipopolysaccharides of gastric and enterohepatic helicobacters. Helicobacter. 2004;9:313-23 pubmed
  8. Pedersen K, Pedersen M, Trøseid M, Gaardbo J, Lund T, Thomsen C, et al. Microbial translocation in HIV infection is associated with dyslipidemia, insulin resistance, and risk of myocardial infarction. J Acquir Immune Defic Syndr. 2013;64:425-33 pubmed publisher
    ..Hence, the gastrointestinal mucosal barrier may be a potential therapeutic target to prevent dyslipidemia and future cardiovascular complications in HIV infection. ..
  9. Mertsola J, Cope L, Munford R, McCracken G, Hansen E. Detection of experimental Haemophilus influenzae type b bacteremia and endotoxemia by means of an immunolimulus assay. J Infect Dis. 1991;164:353-8 pubmed
    ..845, P less than .001). Using antibodies with appropriate specificities, this approach should be useful for rapid detection of a wide range of gram-negative bacteria and endotoxins in blood. ..
  10. Hiki N, Berger D, Dentener M, Mimura Y, Buurman W, Prigl C, et al. Changes in endotoxin-binding proteins during major elective surgery: important role for soluble CD14 in regulation of biological activity of systemic endotoxin. Clin Diagn Lab Immunol. 1999;6:844-50 pubmed
    ..This study demonstrated the role of sCD14 on the bioactivity of circulating endotoxin in a human model of endotoxemia after major abdominal surgery. ..
  11. Trautmann M, Heinemann M, Möricke A, Seidelmann M, Lorenz I, Berger D, et al. Endotoxin release due to ciprofloxacin measured by three different methods. J Chemother. 1999;11:248-54 pubmed
    ..Limulus LPS determinations correlated more precisely with LPS bioactivity than did ELISA values, an observation which underlines the crucial role of LPS determination methods in studies of antibiotic-induced LPS release. ..
  12. Ogawa T. Immunobiological properties of chemically defined lipid A from lipopolysaccharide of Porphyromonas (Bacteroides) gingivalis. Eur J Biochem. 1994;219:737-42 pubmed
    ..toxicity in galactosamine-loaded mice, preparative ability for local Shwartzman reaction, pyrogenicity and Limulus test as compared with compounds 506 and 516...
  13. Hou R. [Treatment of gut-derived endotoxemia with lactulose. An experimental study]. Zhonghua Wai Ke Za Zhi. 1991;29:248-50, 272 pubmed
    ..It is suggested that lactulose can prevent absorbtion of endotoxin from gut and may have an effect on gut-derived endotoxemia. The mechanism of lactulose for treating endotoxemia is discussed. ..
  14. Cohen J. The detection and interpretation of endotoxaemia. Intensive Care Med. 2000;26 Suppl 1:S51-6 pubmed
    ..It may be that the LAL assay, or one of the newer developments, may be more useful in excluding Gram negative infection, but that remains to be shown. ..
  15. Dorn C, Bataille F, Gaebele E, Heilmann J, Hellerbrand C. Xanthohumol feeding does not impair organ function and homoeostasis in mice. Food Chem Toxicol. 2010;48:1890-7 pubmed publisher
    ..Particularly, hepatotoxic effects could be ruled out confirming a good safety profile of xanthohumol as prerequisite for further studies in humans. ..
  16. Jacob S, Nath S, Zade R. Effect of periodontal therapy on circulating levels of endotoxin in women with periodontitis: a pilot clinical trial. Indian J Dent Res. 2012;23:714-8 pubmed publisher
    ..There was significant increase of endotoxins levels (P<.05) immediately after scaling (15 min). SRP can lead to endotoxemia, possibly by release of endotoxins by bacteria of periodontal origin, into the systemic circulation. ..
  17. Lee D, Sung H, Han D, Lee M, Ryu G, Aihara M, et al. In vitro bioassay of endotoxin using fluorescein as a pH indicator in a macrophage cell culture system. Yonsei Med J. 2005;46:268-74 pubmed
    ..The macrophage culture system-method might be a good supplement to the LAL assay for detection of LPS, Gram-negative and Gram-positive bacteria. ..
  18. Sakata M, Inoue T, Todokoro M, Kunitake M. Limulus amebocyte lysate assay for endotoxins by an adsorption method with polycation-immobilized cellulose beads. Anal Sci. 2010;26:291-6 pubmed
    ..This suggests that these compounds inhibit the LAL procedure. By contrast, the adsorption method showed good LPS recovery (88-120%) in all cases, without being inhibited or enhanced by the compounds. ..
  19. Goodin J, Nellis D, Powell B, Vyas V, Enama J, Wang L, et al. Purification and protective efficacy of monomeric and modified Yersinia pestis capsular F1-V antigen fusion proteins for vaccination against plague. Protein Expr Purif. 2007;53:63-79 pubmed
    ..pestis CO92. Thus, vaccination with F1-V monomer and multimeric forms resulted in significant, and essentially equivalent, protection. ..
  20. Kojika M, Sato N, Yaegashi Y, Suzuki Y, Suzuki K, Nakae H, et al. Endotoxin adsorption therapy for septic shock using polymyxin B-immobilized fibers (PMX): evaluation by high-sensitivity endotoxin assay and measurement of the cytokine production capacity. Ther Apher Dial. 2006;10:12-8 pubmed
    ..PMX-DHP is thus considered to be a useful adjuvant therapeutic technique in the treatment of septic shock. Also, PMX-DHP might alleviate the immunosuppression associated with severe sepsis. ..
  21. Luchi M, Morrison D. Comparable endotoxic properties of lipopolysaccharides are manifest in diverse clinical isolates of gram-negative bacteria. Infect Immun. 2000;68:1899-904 pubmed
    ..These relatively minor differences in LPS activity seem unlikely to explain the generally observed discrepancy between serum endotoxin levels and mortality in patients with gram-negative sepsis. ..
  22. Kusumoto S, Fukase K, Fukase Y, Kataoka M, Yoshizaki H, Sato K, et al. Structural basis for endotoxic and antagonistic activities: investigation with novel synthetic lipid A analogs. J Endotoxin Res. 2003;9:361-6 pubmed
    ..The effect of Kdo residues linked to lipid A is also briefly discussed. Molecular dynamics calculations reasonably explain possible conformations required for the biological activity. ..
  23. Szymanska J. Exposure to bacterial endotoxin during conservative dental treatment. Ann Agric Environ Med. 2005;12:137-9 pubmed
    ..Application of a user-friendly water disinfectant to significantly decrease endotoxin concentration in the DUWL water and in the aerosol, is one of recommended methods to reduce health risk. ..
  24. Rønholm E, Tomasdottir H, Runeborg J, Mattsby Baltzer I, Olausson3 M, Aneman A, et al. Gastro-intestinal complement activation during human liver transplantation: impact on postoperative liver function. Acta Anaesthesiol Scand. 2000;44:850-7 pubmed
    ..Gastro-intestinal complement activation may contribute to postoperative parenchymatous liver cell injury and liver dysfunction in patients undergoing liver transplantation. ..
  25. Cardoso M, de Oliveira L, Koga Ito C, Jorge A. Effectiveness of ozonated water on Candida albicans, Enterococcus faecalis, and endotoxins in root canals. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2008;105:e85-91 pubmed publisher
    ..Ozonated water did not neutralize endotoxin. It could be concluded that ozonated water was effective against C. albicans and E. faecalis but showed no residual effect. No activity on endotoxin was observed. ..
  26. Wiedermann C, Kiechl S, Dunzendorfer S, Schratzberger P, Egger G, Oberhollenzer F, et al. Association of endotoxemia with carotid atherosclerosis and cardiovascular disease: prospective results from the Bruneck Study. J Am Coll Cardiol. 1999;34:1975-81 pubmed
  27. Park D, Teschke K, Bartlett K. A model for predicting endotoxin concentrations in metalworking fluid sumps in small machine shops. Ann Occup Hyg. 2001;45:569-76 pubmed
    ..In addition, since there was correlation within-shop, contamination of one sump in a shop may suggest changing the fluids in all. ..
  28. Yoshida K, Niki Y, Mohri K, Miyashita N, Kobashi Y, Oka M, et al. [Evaluation of a new standard beta-glucan CSBG in the measurement of beta-glucan in plasma by alkaline treatment, chromogenic automated kinetic assay]. Kansenshogaku Zasshi. 2004;78:519-21 pubmed
  29. Balagopal A, Gama L, Franco V, Russell J, Quinn J, Higgins Y, et al. Detection of microbial translocation in HIV and SIV infection using the Limulus amebocyte lysate assay is masked by serum and plasma. PLoS ONE. 2012;7:e41258 pubmed
    ..2% plasma and with diazo-coupling was closely correlated between the first and second samples (R?=?0.66, p<0.05). Undiluted serum and plasma mask LPS detection. The extent of MT may be substantially underestimated. ..
  30. Kamiyama S, Teruya Y, Nohara M, Kanazawa K. Impact of detection of bacterial endotoxin in menstrual effluent on the pregnancy rate in in vitro fertilization and embryo transfer. Fertil Steril. 2004;82:788-92 pubmed
    ..The pregnancy rate after IVF-ET was significantly higher in women with an endotoxin level of </=200 pg/mL than in women with an endotoxin level of >200.0 pg/mL. ..
  31. Muhammed K, Ozener Q, Akoglu E. Diagnostic value of effluent endotoxin level in gram-negative peritonitis in CAPD patients. Perit Dial Int. 2001;21:154-7 pubmed
  32. Joshi B, Chatterji A, Bhonde R. Long-term in vitro generation of amoebocytes from the Indian horseshoe crab Tachypleus gigas (Müller). In Vitro Cell Dev Biol Anim. 2002;38:255-7 pubmed
  33. Ohno N, Furukawa M, Miura N, Adachi Y, Motoi M, Yadomae T. Antitumor beta glucan from the cultured fruit body of Agaricus blazei. Biol Pharm Bull. 2001;24:820-8 pubmed
    ..These facts strongly suggest that a highly branched 1,3-beta-glucan segment forms the active center of the antitumor activity...
  34. Gee A, Sumstad D, Stanson J, Watson P, Proctor J, Kadidlo D, et al. A multicenter comparison study between the Endosafe PTS rapid-release testing system and traditional methods for detecting endotoxin in cell-therapy products. Cytotherapy. 2008;10:427-35 pubmed publisher
    ..The Endosafe system provided reliable results with products typically produced in cell-therapy manufacturing facilities, and would be an appropriate test on which to base the release of time-sensitive cell-therapy products. ..
  35. Perdomo Morales R, Pardo Ruiz Z, Spreitzer I, Lagarto A, Montag T. Monocyte activation test (MAT) reliably detects pyrogens in parenteral formulations of human serum albumin. ALTEX. 2011;28:227-35 pubmed
    ..The MAT correlates with the rabbit pyrogen test, providing a higher safety level for pyrogenicity testing of HSA and probably other therapeutic proteins. ..
  36. Torres A, Mas Moruno C, Perez Paya E, Albericio F, Royo M. Trivalent PEGylated platform for the conjugation of bioactive compounds. Bioconjug Chem. 2011;22:2172-8 pubmed publisher
    ..These results prove the efficacy of this trivalent platform as a multivalent ligand scaffold for biological applications. ..
  37. Onodera C, Takahashi G, Kan S, Shozushima T, Matsumoto N, Inada K, et al. Experimental application of a synthetic luminescent substrate assay using endotoxin-specific limulus amebocyte lysate to human blood. J Infect Chemother. 2012;18:370-7 pubmed publisher
    ..However, the shortest time after collecting the specimen in which it was possible to make measurements was 30-40 min, suggesting that if an assay is established, it will be possible to use the method as a novel blood endotoxin assay. ..
  38. Hansen L, Poulsen O, Wurtz H. Endotoxin potency in the A549 lung epithelial cell bioassay and the limulus amebocyte lysate assay. J Immunol Methods. 1999;226:49-58 pubmed
    ..The lack of correlation when organic dust samples are tested may reflect the fact that these samples contain biological active compounds, which are non-reactive in the LAL-assay but stimulate IL-8 secretion from epithelial cells. ..
  39. Imamura S, Nakamizo M, Kawanishi M, Nakajima N, Yamamoto K, Uchiyama M, et al. Bovine whole-blood culture as a tool for the measurement of endotoxin activities in Gram-negative bacterial vaccines. Vet Immunol Immunopathol. 2013;153:153-8 pubmed publisher
    ..b>Limulus test revealed that endotoxin activity was remarkably reduced when aluminum hydroxide gel was added to LPS...
  40. Haishima Y, Murai T, Nakagawa Y, Hirata M, Yagami T, Nakamura A. Chemical and biological evaluation of endotoxin contamination on natural rubber latex products. J Biomed Mater Res. 2001;55:424-32 pubmed
    ..These findings clearly showed that the pyrogenicity found in these latex products originated from endotoxins contaminating the products. ..
  41. Singh U, Levin L, Grinshpun S, Schaffer C, Adhikari A, Reponen T. Influence of home characteristics on airborne and dustborne endotoxin and ?-D-glucan. J Environ Monit. 2011;13:3246-53 pubmed publisher
    ..Furthermore, even though statistically significant, the correlations between airborne and dustborne contaminants were weak. This indicates that airborne concentrations cannot be reliably predicted based on dustborne concentrations. ..
  42. Miyazaki T, Kohno S, Mitsutake K, Maesaki S, Yamada H, Yasuoka A, et al. [Limulus test (factor G) and polysaccharides from fungus]. Kansenshogaku Zasshi. 1992;66:1030-6 pubmed
    Picogram quantifies per ml of endotoxin showed a positive limulus test. The concentration more than ten pg/ml of curdlan ((1-3)-beta-D-glucan) was also positive to a conventional limulus test, which contains a factor G...
  43. Aroutcheva A, Ling Z, Faro S. Prevotella bivia as a source of lipopolysaccharide in the vagina. Anaerobe. 2008;14:256-60 pubmed publisher
  44. Lao H, Lin Q, Yang M, Liang X, Shi W, Li H, et al. [Kinetic turbidimetric limulus assay of bacterial endotoxin in the replacement solution of on-line hemodiafiltration]. Nan Fang Yi Ke Da Xue Xue Bao. 2006;26:242-4 pubmed
    ..endotoxin in the replacement solution of on-line hemodiafiltration (on-line HDF) using kinetic turbidimetric limulus test. Validation test was performed with the replacement solution of on-line HDF in which quantified standard ..
  45. Ogikubo Y, Norimatsu M, Noda K, Takahashi J, Inotsume M, Tsuchiya M, et al. Evaluation of the bacterial endotoxin test for quantification of endotoxin contamination of porcine vaccines. Biologicals. 2004;32:88-93 pubmed
    ..Our findings suggest that the bacterial endotoxin test using endotoxin-specific Limulus amoebocyte lysate (LAL) can detect endotoxin contamination in commercial porcine vaccines containing either oil or aluminum adjuvants. ..
  46. Mueller M, Lindner B, Kusumoto S, Fukase K, Schromm A, Seydel U. Aggregates are the biologically active units of endotoxin. J Biol Chem. 2004;279:26307-13 pubmed
    ..We further confirmed this result by preparing monomeric lipid A and LPS by a dialysis procedure and found that, at the same concentrations, only the aggregates were biologically active, whereas the monomers showed no activity. ..
  47. Chang H, Sack D. Detection of anti-lipopolysaccharide antibodies to Vibrio cholerae O1 and O139 using a novel microtiter limulus amebocyte lysate (LAL) assay. Clin Chim Acta. 2001;312:49-54 pubmed
    ..This study showed that antibodies to V. cholerae blocked the LAL reaction and suggested that the LAL blocking assay could be applicable for determining antibodies to other endotoxin-containing bacteria and microorganisms. ..
  48. Massaldi H, Morais V. Evolution of endotoxin contamination during production of a therapeutic serum. PDA J Pharm Sci Technol. 2007;61:375-82 pubmed
    ..Only with proper sanitization of material, and by applying the caprylic acid purification process with a starting plasma below 4-5 EU/mL, would it be possible to achieve a final product within the norm. ..
  49. Kataoka M, Ochiai M, Yamamoto A, Horiuchi Y. A need for careful evaluation of endotoxin contents in acellular pertussis-based combination vaccines. Biologicals. 2012;40:49-54 pubmed publisher
    ..As an adequate in vitro test method could not be identified for controlling the safety of the interfering batch, an appropriate in vivo test would be required for testing such vaccines. ..
  50. Andrä J, Garidel P, Majerle A, Jerala R, Ridge R, Paus E, et al. Biophysical characterization of the interaction of Limulus polyphemus endotoxin neutralizing protein with lipopolysaccharide. Eur J Biochem. 2004;271:2037-46 pubmed
    ..Thus, LPS-ENP complexes might enter target membranes of immunocompetent cells, but are not able to activate due to the ability of ENP to change LPS aggregates from an active into an inactive form. ..
  51. Becker K, Ditter B, Nimsky C, Urbaschek R, Urbaschek B. [Endotoxin contents of phytopharmaceuticals: correlation with clinically observed side effects]. Dtsch Med Wochenschr. 1988;113:83-7 pubmed
    ..These findings underline the urgent need for a widening of the regulations on testing for pyrogens to include those parenteral preparations which now do not have to be tested because of their small volume (less than 15 ml)...
  52. de Souza L, Koury M. Chemical and biological properties of endotoxin from Leptospira interrogans serovars canicola and icterohaemorrhagiae. Braz J Med Biol Res. 1992;25:467-75 pubmed
    ..3, 5.6 and 1.5 for canicola. 2. The preparations presented several biological activities: positive Limulus test (1.0 pg/ml) for icterohaemorrhagiae and canicola PCP extract and 0.5 pg/ml for E...
  53. Shimizu T, Obata T, Sonoda H, Akabori H, Miyake T, Yamamoto H, et al. Diagnostic potential of endotoxin scattering photometry for sepsis and septic shock. Shock. 2013;40:504-11 pubmed publisher
    ..In conclusion, compared with turbidimetric LAL assay, ESP more sensitively detected plasma endotoxin and significantly discriminated between sepsis and septic shock in patients undergoing gastrointestinal emergency surgery. ..