Rapid presumptive diagnosis of gonococcal cervitis by the limulus lysate assay

Spagna, Vincent A.; Prior, Richard B.; Perkins, Robert L.

doi:10.1016/0002-9378(80)90702-4

Cited by 18 publications

(7 citation statements)

References 15 publications

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“…The concept involved using a lysate made from the washed amoebocytes of the horseshoe crab (Limuluspolyphemus) which forms a gel in the presence of small quantities (nanograms) of bacterial endotoxin. The lysate was shown to be extremely sensitive to intact outer membrane components of N. gonorrhoeae (13) and more sensitive to this organism than other gram-negative bacteria tested (18). When urethral exudates, properly diluted according to lysate sensitivity, were mixed with lysate and incubated, gelation occurred when N. gonorrhoeae was present.…”

Section: Discussionmentioning

confidence: 99%

Application of a Limulus test device in rapid evaluation of gonococcal and nongonococcal urethritis in males

Prior¹,

Spagna²

1981

J Clin Microbiol

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A test device incorporating Limulus amoebocyte lysate (Mallinckrodt, Inc., St. Louis, Mo.) was developed for the rapid, presumptive diagnosis of gonococcal and nongonococcal disease in males. The device, which was evaluated in 550 men with exudative urethritis, consisted of a specimen collection syringe, a dilution reservoir containing 10 ml of pyrogen-free water, and a Limulus amoebocyte lysate singletest vial. After specimen collection, the syringe was affixed to the dilution reservoir for rapid, accurate dilution of the clinical sample. Contamination of the specimen and potential biohazards to the user were prevented. The diluted sample was then transferred (via the collection syringe) to the lysate test vial for assay of endotoxin. Various incubation times at 37°C were also studied in an additional 301 male patients, and time was reduced from the standard 60 to 30 min while still retaining equivalent predictability of culture results (P > 0.05). Of the 550 males evaluated with the test device, 366 had positive cultures for Neisseria gonorrhoeae, and 184 were negative. A sensitivity of 99.2% and a specificity of 96.7% were obtained with the test device. Overall ability to predict culture results was 98.4%. Gram-stain sensitivity and specificity were 96.4% and 99.5%, respectively, with an overall accuracy of 97.5%. There were no statistical differences between the Limulus amoebocyte lysate test and Gram stain in predicting cultures (P > 0.05). Thus, use of the Limulus amoebocyte lysate test device would enable the private physician to make an accurate, presumptive diagnosis of gonococcal and nongonococcal disease in males with exudative urethritis within 30 min without the need of a microscope and to initiate proper therapy during the patient's initial evaluation.

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Section: Discussionmentioning

confidence: 99%

Application of a Limulus test device in rapid evaluation of gonococcal and nongonococcal urethritis in males

Prior¹,

Spagna²

1981

J Clin Microbiol

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“…The test for this reaction is known as the Limulus amoebocyte lysate (LAL) test and currently constitutes the most sensitive method available for detection of bacterial endotoxin. The LAL test has been shown to have potential diagnostic value for the rapid screening of a variety of body fluids for gram-negative infectious disease (13)(14)(15)(16)(17)(26)(27)(28) and has been adopted for endotoxin detection in pharmaceutical drugs and medical devices (12,21,31). The unique properties of LAL include its sensitivity to and reported specificity for endotoxin; however, specificity remains a controversial issue as a result of reports of LAL activation (gelation) by substances other than endotoxin (1,5,6,11,29,34).…”

mentioning

confidence: 99%

Establishment of beta-hydroxy fatty acids as chemical marker molecules for bacterial endotoxin by gas chromatography-mass spectrometry

Maitra¹,

Nachum²,

Pearson³

1986

Appl Environ Microbiol

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Selected ion-monitoring gas chromatography-mass spectrometry was used for detection of beta-hydroxy fatty acids as an independent assay for the presence or absence of endotoxin in materials claimed to induce nonspecific activation of Limulus amoebocyte lysate. To this end, suspensions of gram-negative and-positive bacteria, one fungal species, cerebrospinal fluid, and hollow-fiber hemodialyzer rinses were assayed for endotoxin by gas chromatography-mass spectrometry and the Limulus amoebocyte lysate assay. Good qualitative agreement was shown for both methods when suspensions of test organisms were assayed. Two false-negative results were obtained by gas chromatography-mass spectrometry assays of cerebrospinal fluid and were shown to be a result of insufficient endotoxin in the cerebrospinal fluid specimens for detection by gas chromatography-mass spectrometry. Hemodialyzer rinses were Limulus assay positive; however, no betahydroxy fatty acids were detected by gas chromatography-mass spectrometry. These data were compared with data obtained from USP rabbit pyrogen tests of the rinse materials (nonpyrogenic) and chemical characterization of the Limulus assay-reactive rinses, which showed the rinses to be cellulosic in nature. It is suggested that beta-hydroxy fatty acids, as assayed by selected ion-monitoring gas chromatography-mass spectrometry, be used as chemical marker molecules for the presence or absence of endotoxin in materials reported to cause nonspecific activation of Limulus amoebocyte lysate.

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“…Aqueous lysates prepared from the circulating cells (amoebocytes) of the horseshoe crab (Limulus polyphemus) gel when exposed to minute amounts of bacterial endotoxin (11). This reaction is the basis of the Limulus amoebocyte lysate (LAL) test, which has been shown to have potential usefulness for rapid diagnosis of gram-negative infections in various body fluids, including cerebrospinal fluid (8,16), urine (17), and urethral and cervical exudates for the presumptive diagnosis of gonorrhea (15,20,24,25).…”

mentioning

confidence: 99%

Chromogenic Limulus amoebocyte lysate assay for rapid detection of gram-negative bacteriuria

Nachum¹,

Berzofsky²

1985

J Clin Microbiol

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A chromogenic Limulus amoebocyte lysate assay was evaluated as a rapid screening test for the detection of clinically significant gram-negative bacteriuria. The development of a distinctive yellow color after the addition of chromogenic substrate to the Limulus amoebocyte lysate-urine reaction mixture was used to measure 2105 gram-negative bacteria per ml. A total of 324 urine specimens were assayed, with 68 gram-negative urinary tract infections identified as defined by quantitative urine colony counts of 2105 bacteria per ml. Of these, 68 and 67 of 68 were detected by the chromogenic Limulus amoebocyte lysate assay at urine dilutions of 1:10 and 1:20, respectively. Nine false-positive chromogenic Limulus amoebocyte lysate assay results were observed at both urine dilutions and in the same specimens. At a urine dilution of 1:10, sensitivity and specificity were 100 and 96.6%, respectively, with predictive values of 100% for a negative test and 88.3% for a positive test. At a urine dilution of 1:20, sensitivity and specificity were 98.6 and 96.6%, respectively; predictive values were 99.6% for a negative test and 88.3% for a positive test. These data suggest that chromogenic Limulus amoebocyte lysate assay of urine has potential usefulness as a rapid, reliable, and easily performed and interpreted screening test for the diagnosis of clinically significant gram-negative bacteriuria.

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Rapid presumptive diagnosis of gonococcal cervitis by the limulus lysate assay

Cited by 18 publications

References 15 publications

Application of a Limulus test device in rapid evaluation of gonococcal and nongonococcal urethritis in males

Application of a Limulus test device in rapid evaluation of gonococcal and nongonococcal urethritis in males

Establishment of beta-hydroxy fatty acids as chemical marker molecules for bacterial endotoxin by gas chromatography-mass spectrometry

Chromogenic Limulus amoebocyte lysate assay for rapid detection of gram-negative bacteriuria

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