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PubMed1977

Detection of endotoxin in cerebrospinal and joint fluids by limulus assay.

Cerebrospinal fluid (CSF) and joint fluid from 81 patients were studied for the presence of endotoxin using the limulus assay. All fluids that grew Gram-negative organisms had a positive limulus test. However, one sample of CSF and one of joint fluid, both very purulent, that contained Gram-positive organisms also gave positive limulus assays. All culturally negative fluids had a negative limulus assay. Thus, a positive limulus assay with CSF or joint fluid is indicative of bacterial infection probably caused by Gram-negative organisms and can be an extremely useful diagnostic adjunct.

Adult↗
PubMed1983

[Severe surgical infection and DIC].

Severe surgical infection is one of the most obvious clinical settings of DIC. Although many initiating triggers have been discussed, the most potent might be a bacterial endotoxin which activates Hageman factor or some components of serum complement. We examined a disorder of blood coagulation system in severe surgical infection in terms of the results of limulus test for detection of circulating endotoxin. In 59 of such patients, 10 cases (32.3%) of DIC diagnosed by MINNA 's criteria were recognized in 31 limulus positive cases, whereas only 4 cases (14.3%) in 28 limulus negative groups. Each clinical course, prognosis, or the relationship with MOF were also discussed in detail. It was emphasized that the first requirement for diagnosis of DIC should be awareness of its possibility and careful observation of platelet counts. The immediate needs for treatment of DIC are (1) a cure of underlying infection by surgical drainage and chemotherapy, and (2) inhibition of intravascular coagulation by use of heparin and/or FOY etc. Our experiments performed by a continuous drop infusion of both 0.2 mg/kg 055; B5 E. Coli endotoxin and several doses of FOY for 4 hours into rabbits showed that the most potent inhibitory effect of FOY on platelet and WBC was observed in 0.1 mg/kg/min.

Adult↗
PubMed1977

Statistical determination of endotoxin content in influenza virus vaccine by the limulus amoebocyte lysate test.

To determine laboratory-to-laboratory variability of the limulus amoebocyte lysate (LAL) test for evaluating endotoxin content in influenza virus vaccine, a collaborative study was designed. Participants were six influenza virus vaccine manufacturers and the Bureau of Biologics (BoB). Lysate lot 116, Reference Influenza Virus Vaccine for Endotoxin Assay E-1, and four test vaccines having different ratios of LAL activity relative to reference E-1 were supplied by the BoB. Each laboratory used its normal test procedure. All vaccines were coded. One pair (reference and test) of vaccines per day was tested for 4 days a week over a 4-week period. All data were analyzed at the BoB. The degree of variability experienced by testing laboratories was estimated by the study. This estimate did not conflict with experience gained from previous routine testing in any of the participating laboratories. A statistical approach to the evaluation of LAL data from testing influenza virus vaccine for endotoxin content was developed based upon the overall variation obtained from the collaborative study.

Drug Contamination↗
PubMed1993

[The effect of mistletoe lectins on the limulus amebocyte lysate test].

Former reports about too high and from batch to batch changing endotoxin contents in mistletoe preparations and phytopharmaceuticals for parenteral administration correlated with clinically observed side effects led to investigations of the endotoxin content of ABNOBA-viscum. For that purpose the endotoxin levels of the raw materials, the equipment and the production steps were observed by using the limulus amebocyte lysate (LAL) test. The mean value found in the preparation containing the highest available concentration (dilution level 2 containing 15 mg of plant extract from 20 mg of fresh mistletoe in 1 ml) came to 66.7 endotoxin units (EU) per ml, corresponding to about one fifth of the commonly accepted limit value of 350 EU/ml, the human pyrogenic dose. Depending on their dilution level, lower concentrated preparations led also to lower LAL test results. It could be proved that lectins contained in mistletoe preparations cause false positive LAL tests. Such results implicate false too high contents of endotoxin. Different lysates, i.e. Limusate, Pyrogel, Pyrogent, and Pyroquant showed approximately the same results. Microbiological and LAL test results indicate that only 10% of the total amount of the LAL test result was caused by endotoxins. So it is necessary to form a new estimation of the former results of determination of the endotoxin content of mistletoe preparations.

Animals↗
PubMed1976

Remarks on the investigation of sodium nucleinate by means of the limulus amebocyte lysate test.

Results of investigations of sodium nucleinate by means of the limulus amebocyte lysate (LAL) test are described. It was found that the sensitivity of this new pyrogen test to bacterial lipopolysaccharides is nearly the same as to sodium nucleinate, which is prescribed as pyrogen standard in the Pharmacopeia of the German Democratic Republic and recommended as such in the Compendium Medicamentorum.

Animals↗
PubMed1983

[Studies on metabolism of fat emulsion in endotoxemic dogs].

Changes of fat metabolism in endotoxemic dogs were studied by means of intravenous fat tolerance test. Twelve dogs subjected to the present studies were divided into two groups; peritonitis was produced in one group (6 dogs) and 1.5 mg/kg of E. coli lipopolysaccharide was injected intravenously in another groups (6 dogs). After production of peritonitis, Limulus test was positive in all of the six dogs, however there were no significant changes in blood sugar levels, lactate/pyruvate ratio, and elimination rate of fat emulsion from the venous blood (K-value) after the induction of endotoxemia by peritonitis. On the other hand, a significant fall of the blood sugar levels and an increase of lactate/pyruvate ratio were found after the injection of endotoxin. A significant fall of K-value (from 0.0133 +/- 0.0056 to 0.0069 to 0.0024) was also observed in these six dogs. Results of the present studies suggest that the metabolism of fat emulsion is significantly affected by the intravenous injection of endotoxin, but not by the production of peritonitis if there is no disturbance of tissue perfusion.

Animals↗
PubMed1979

Determining endotoxin content of ground beef by the Limulus amoebocyte lysate test as a rapid indicator of microbial quality.

Eighty-four samples of ground beef were placed into five half-log cycle groups based upon aerobic plate count (APC) results. Endotoxins were determined by the Limulus amoebocyte lysate test (LAL), and gram-negative viable counts were determined by a violet red bile agar overlay method. Ten samples with a log of APC of less than 5.50 had an APC mean of less than 5.24 and mean endotoxin content by the LAL of 51 ng/g. The 15 samples with APCs between a log of 5.50 and 5.99 had an APC mean of 5.79/g and an endotoxin mean of 103.8 ng/g. Twenty-eight samples had APCs between a log of 6.00 and 6.49 with a mean of 5.28/g and an endotoxin mean of 1106.4 ng/g. The 20 samples with APCs between a log of 6.50 and 7.00 had a mean of 6.77/g and an endotoxin mean of 5067.6 ng/g, while 11 samples had a log of APCs of greater than 7.00 with a mean of 7.53 and an endotoxin mean of 7,472 ng/g. Correlation of half-log cycle mean APC and violet red bile agar counts with mean endotoxin content were both highly significant, indicating that LAL-determined endotoxin content can be used to make a rapid approximation of viable plate counts. Because results can be obtained by LAL in 1 h, the finding of low levels of endotoxins can be taken to indicate low-count meat. The use of additional tests of microbial quality may be necessary when high endotoxin levels are found because the LAL detects both viable and nonviable cells.

Aerobiosis↗
PubMed2005

[Limulus amebocyte lysate test use in clinical diagnostic in children with infectious diseases--preliminary report].

OBJECTIVE: The aim of this preliminary study was (1) to evaluate the usefulness of the Limulus amebocyte lysate (LAL) test as a diagnostic tool in pediatric clinic, and (2) to examine the serum ET levels in hospitalized patients. MATERIAL AND METHODS: Bacterial blood endotoxin (ET) was measured in 50 children with infections of different etiology and localization. A Limulus amebocyte lysate (LAL) test was used. CONCLUSIONS: The usefulness of LAL test in routine diagnostic procedures in hospitalized children with infections was not proven so far. However, the LAL test appeared useful in the urinary tract infection (UTI) cases in which significant elevation of serum ET levels were demonstrated. Moreover, it was confirmed that LAL test can be performed as well in serum: the mean values of serum ET levels in our patients were comparable with published ET plasma values in adults.

Animals↗
PubMed1990

[Development of an endotoxin-specific Limulus amebocyte lysate test blocking beta-glucan-mediated pathway by carboxymethylated curdlan and its application].

We developed a simple new endotoxin-specific assay method that uses Limulus amebocyte lysate (LAL) containing a sufficient amount of a water-soluble (1----3)-beta-D-glucan derivative as a blocker of the (1----3)-beta-D-glucan-mediated coagulation pathway. The addition of 0.1 mg/ml or more of carboxymethylated (1----3)-beta-D-glucan completely blocked the activation of LAL by (1----3)-beta-D-glucan itself. The assay of endotoxin was unaffected by the presence of 1 mg/ml carboxymethylated (1----3)-beta-D-glucan. Spiked endotoxin was recovered well from beta-glucans by the turbidimetric kinetic method with LAL containing 1 mg/ml of carboxymethylated (1----3)-beta-D-glucan. Besides, this new LAL formulation was applied for an endotoxin-specific assay by the conventional gel-clot method or the chromogenic method. Gram-negative bacteria were specifically detected by the turbidimetric kinetic method with the LAL formulation. This LAL formulation may be used for an endotoxin-specific assay not only in pharmacology but also in clinical microbiology.

Bacteria↗
PubMed1976

Use of Limulus lysate for detecting gram-negative neonatal meningitis.

The Limulus lysate test was evaluated as a method for rapid detection of neonatal gram-negative bacterial meningitis. A total of 208 CSF samples were collected from 145 newborn infants suspected of having meningitis. Initial samples from all six babies with culture-proven gram-negative bacterial meningitis had positive Limulus tests within 30 minutes of incubation. Samples from 14 infants with gram-negative organisms isolated only in blood and/or urine as well as from four neonates with gram-positive organisms in CSF cultures and from 13 with gram-positive organisms in blood cultures all yielded negative Limulus tests. Thus, of 37 newborn infants with bacteria demonstrated in systemic cultures, only those six with gram-negative organisms in CSF had positive CSF Limulus tests. The CSF Limulus test was shown to be a rapid, reliable, and specific test for the detection on neonatal gram-negative bacterial meningitis.

Arachnida↗