Rapid Detection of Gram-Negative Bacterial Meningitis by the Limulus Lysate Test

Nachum, R; Lipsey, Allen I.; Siegel, Stuart E.

doi:10.1056/nejm197311012891801

Cited by 103 publications

(27 citation statements)

References 22 publications

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“…A correctly performed LAL assay can detect approximately 103 gram-negative bacteria per ml of specimen (149). Nachum reviewed 4,884 CSF specimens which had been examined by LAL assays and calculated the overall sensitivity and specificity of the LAL assay (98 …”

Section: Concentrationmentioning

confidence: 99%

Laboratory diagnosis of bacterial meningitis

1992

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Bacterial meningitis is relatively common, can progress rapidly, and can result in death or permanent debilitation. This infection justifiably elicits strong emotional reactions and, hopefully, immediate medical intervention. This review is a brief presentation of the pathogenesis of bacterial meningitis and a review of current knowledge, literature, and recommendations on the subject of laboratory diagnosis of bacterial meningitis. Those who work in clinical microbiology laboratories should be familiar with the tests used in detecting bacteria and bacterial antigens in cerebrospinal fluid (CSF) and should always have the utmost appreciation for the fact that results of such tests must always be reported immediately. Academic and practical aspects of the laboratory diagnosis of bacterial meningitis presented in this review include the following: anatomy of the meninges; pathogenesis; changes in the composition of CSF; etiological agents; processing CSF; microscopic examination of CSF; culturing CSF; methods of detecting bacterial antigens and bacterial components in CSF (counter-immunoelectrophoresis, coagglutination, latex agglutination, enzyme-linked immunosorbent assay, Limulus amebocyte lysate assay, and gas-liquid chromatography); use of the polymerase chain reaction; and practical considerations for testing CSF for bacterial antigens.

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Section: Concentrationmentioning

confidence: 99%

Laboratory diagnosis of bacterial meningitis

1992

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“…terial endotoxin may contribute to the PG-stimulating activity by RSF: (i) Preparations of active RSF gave negative Limulus assays for endotoxin (13).…”

Section: Resultsmentioning

confidence: 99%

Mechanism of stimulation of prostaglandin synthesis by a factor from rheumatoid synovial tissue.

Robinson¹,

Bastian²,

Hamer³

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

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[1-14C]AA levels in cultures of untreated cells. These observations are inconsistent with a mechanism of prostaglandin stimulation consisting of an increase in phospholipase activity, because this mechanism would increase free AA levels as well as prostaglandins. A mechanism is proposed in which free AA is maintained at low steady-state levels by reacylation of free AA into phospholipids at a rate more rapid than its reaction with cyclooxygenase to form prostaglandins. In this mechanism, stimulation ofthe rate ofthe cyclooxygenase step by RSF accounts for increased prostaglandin synthesis as well as the decreased release of AA. On the basis of data previously reported by others, it is suggested that this mechanism may also be applicable to the stimulation of prostaglandin synthesis by several other agents. Preliminary characterization of the RSF indicates that it is a protein, and molecular seive chromatography indicates that its molecular weight is about 18,000. The production of RSF by rheumatoid synovial tissue is suppressed to undetectable levels by 1 F.M dexamethasone.It has been generally believed that the rate-limiting step in prostaglandin (PG) synthesis is the phospholipase-catalyzed hydrolysis of phospholipids and that the mechanisms of action ofvarious factors stimulating PG synthesis involve the activation of phospholipases. These hypotheses have been in part based on the facts that the quantities of free arachidonic acid (AA) within cells and tissue are insufficient to account for quantities of PGs synthesized and that stimulation of PG synthesis may be accompanied by variable increases in the concentrations of free AA (1-6).We report here the results ofstudies ofsome ofthe properties and the mechanism of action of a PG-stimulating factor produced by rheumatoid synovial tissue (RSF). In these experiments, the stimulation of PGE2 synthesis is generally accompanied by decreases in free AA concentrations. We conclude that these observations are inconsistent with a mechanism based on stimulation of phospholipase activity. A mechanism is proposed in which RSF stimulates the rate of the cyclooxygenase step. A preliminary report of some of this has appeared (7).MATERIALS AND METHODS Rheumatoid Synovial Cell Strains. Cells were grown from explants ofrheumatoid synovial tissue by using methods similar to those described by others (8, 9). Confluent cells were passaged using 0.05% trypsin/0.02% EDTA solution (GIBCO no. 530), and new plates were seeded at a density of 1.9 X 105 cells growing to =7 X 106 per 60-mm plate when confluent. For some experiments cells were passaged into multiwell plates at a similar density. Cells were stored in liquid N2 in a freezing mixture of minimal essential medium with 25% fetal bovine serum and 8% dimethyl sulfoxide. Cells were used for these experiments after 4 to 10 passages; no dependence of PG production on passage number was observed.Rheumatoid Synovial Explant Cultures. Explant cultures of rheumatoid synovial tissue were prepared (10). Approximately 60 mg of tissue (wet...

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“…Assuming that in shelf water a similar distribution and concentration of bacteria·in sediments and the water column occurs and assuming a water depth of 100 meters it can be estimated that under one square To study ·the correlation between cell number and rates of (Levin and Bang, 1964) it has been employed for the detection of endotoxemia (Levin et al , 1972;Levin, 1973), bacteriuria (Jorgensen et al, 1973) and for the diagnosis of Gram-negative spinal meningitis (Nachum et al, 1973 Boston. In this study we will study the incidence and the severity of endotoxemia in children with severe cases of diarrhea.…”

Section: -3-mentioning

confidence: 99%

Marine microbiology. Progress report, 1973--1974

Watson¹

1974

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Rapid Detection of Gram-Negative Bacterial Meningitis by the Limulus Lysate Test

Cited by 103 publications

References 22 publications

Laboratory diagnosis of bacterial meningitis

Laboratory diagnosis of bacterial meningitis

Mechanism of stimulation of prostaglandin synthesis by a factor from rheumatoid synovial tissue.

Marine microbiology. Progress report, 1973--1974

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