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PubMed1990

A rapid latex agglutination assay for the determination of plasma fibrinogen.

A monoclonal antibody to intact fibrinogen has been employed to develop a rapid latex agglutination assay for the estimation of plasma fibrinogen. The monoclonal antibody, 45J, recognizes an epitope located in the mid-section of the carboxy terminal end of the A alpha-chain. The epitope is destroyed by plasmin digestion of fibrinogen and there is no immunoreactivity with soluble cross-linked fibrin degradation products. The latex agglutination assay developed with the antibody is unaffected by aprotinin or anticoagulants, such as citrate, heparin or EDTA. When this method was compared with functional clotting assays, excellent correlation was observed with normal and pathological samples. After sample collection and dilution, the assay takes just two minutes to complete. Therefore, this procedure offers a simple and rapid assay for the measurement of intact fibrinogen in plasma.

Animals↗
PubMedDate not supplied

Evaluation of a latex particle agglutination kit in pneumococcal disease.

Latex particle agglutination for Streptococcus pneumoniae was evaluated in 76 patients. Fifteen of these patients had invasive disease due to S. pneumoniae including 12 with meningitis, 2 with occult bacteremia and 1 with suppurative arthritis. Five of the patients with meningitis also had bacteremia. Pneumococcal antigen was detected in the cerebrospinal fluid of 9 of the 12 patients with meningitis (sensitivity 75%). However, antigen was detected in the serum of only two of the six patients with bacteremia (sensitivity 33%) and was detected in the urine of none of five patients with bacteremia (sensitivity 0%). Consequently latex particle agglutination appears to be useful when cerebrospinal fluid is examined in patients with pneumococcal meningitis but does not appear to be sufficiently sensitive to warrant its use with serum or urine in patients with invasive disease due to S. pneumoniae. The specificity of the system used here appeared satisfactory, since pneumococcal antigen was not detected in any of the body fluids from the 61 patients without evidence of invasive pneumococcal disease (specificity 100%).

Antigens, Bacterial↗
PubMedDate not supplied

Clinical performance and effect on treatment rates of latex agglutination testing for streptococcal pharyngitis in an emergency department.

Of 556 children with symptoms of streptococcal pharyngitis in a pediatric hospital emergency department, 38.8% had positive results on either a rapid latex agglutination test or throat culture for Group A Streptococcus. Twenty-six percent of patients with negative latex agglutination test results had growth on culture of a paired specimen. Under the best assumption rapid test sensitivity was 44.9%. The treatment rate (84%) for cases detected by the rapid test was markedly higher than the rate (44%) for cases detected only by culture (P less than 0.001), presumably because the rapid test result is available sooner. We conclude that: the rapid test had a lower sensitivity in routine use in our pediatric emergency department population than in previous reports from research settings; and using the rapid test in addition to culture can significantly increase treatment rates.

Adolescent↗
PubMed1989

Evaluation of a latex agglutination test for Rickettsia conorii antibodies in seropositive patients.

A latex agglutination test for antibodies to Rickettsia conorii was compared with micro-immunofluorescence (the reference test for total antibodies); 179 sera were from 115 confirmed cases of Mediterranean Spotted Fever, and 101 were from pregnant women (control group) who had no detectable antibodies by the reference method. The micro-immunofluorescence test for specific IgM antibodies was used to clarify some discordant results. The agreement obtained between latex-R. conorii and micro-immunofluorescence was 95%. Sensitivity and specificity were 96% and 93% respectively. When micro-immunofluorescence results for specific IgM antibodies were included, these figures rose to 96 and 99%, and agreement was almost 97%. Latex agglutination is a simpler and more rapid technique than micro-immunofluorescence and is suitable for the screening as well as for the titration of R. conorii antibodies.

Antibodies, Bacterial↗
PubMed1994

Sexual agglutination substances require a 'carrier' glycoprotein for integration into the cell wall of Saccharomyces cerevisiae.

Sexual agglutination, caused by agglutination substance (AS) on a and alpha cell walls, is the first indispensable step of the mating reaction in ascosporogenous yeasts including Saccharomyces cerevisiae. The AS biosynthetic process in S. cerevisiae was investigated by pulse label-chase experiments with analysis by polyacrylamide gel electrophoresis (PAGE) for 16 h in the presence of urea. Because of its low mobility, AS can be separated from other proteins by prolonged PAGE. Nascent AS was integrated into cell walls after it linked covalently to a 'carrier' glycoprotein. The results suggest that the 'carrier' is synthesized stepwise through three distinct precursors (III-->II-->I). The 'carrier' glycoprotein (I) and its precursors (II, III) were synthesized in both a, alpha haploid and a/alpha diploid cells. The N-glycosylation linkage inhibitor, tunicamycin, and protein synthesis inhibitor, puromycin, inhibited the III to I maturation. The results indicated that both the 'carrier' and the nascent active site of AS linked to the 'carrier' are integrated into the wall in a haploid cell while the 'carrier' alone is integrated in a diploid cell.

Agglutinins↗
PubMed1996

Aeromonas trota strains, which agglutinate with Vibrio cholerae O139 Bengal antiserum, possess a serologically distinct fimbrial colonization factor.

Pili of Aeromonas trota strain 1220, which agglutinates with Vibrio cholerae O139 Bengal antiserum, were purified and characterized. The molecular mass of the subunit protein was estimated to be 20 kDa and the pl was 5 center dot 4. The pili were immunologically unrelated to the other Aeromonas pili reported so far. However, the N-terminal amino acid sequence of the subunit pilin was similar to those of the pilins from other Aeromonas pili reported previously. Neither A. trota cells nor pili purified from strain 1220 agglutinated human and rabbit erythrocytes, but both adhered to the rabbit intestine. Bacterial cells pretreated with antipilus antibody (Fab portion) failed to adhere to the rabbit intestine. Moreover, bacteria did not adhere to the rabbit intestine pretreated with the purified pili. This pilus antigen was not detected in V. cholerae O139 Bengal and other Aeromonas spp. These findings suggest that the pilus of the A. trota strain is a novel colonization factor of Aeromonas spp.

Aeromonas↗
PubMed1978

Plant protoplast agglutination by lectins.

Concanavalin A, soybean (Glycine max L.) lectin, castor bean (Ricinus communis L.) lectin, and peanut (Arachis hypogaea L.) lectin were able to agglutinate protoplasts prepared from a variety of plant species. The seven other lectins tried were unable to agglutinate those protoplasts tested. Protoplasts prepared from 11 species were used. The lectins examined were not able to differentiate among protoplasts of different species.

Journal Article↗
PubMed1987

Early diagnosis of acute myocardial infarction with a rapid latex agglutination test for semi-quantitative estimation of serum myoglobin.

A rapid latex agglutination test for the detection of elevated levels of myoglobin in serum was evaluated in a prospective study of 236 patients consecutively admitted to a Coronary Care Unit on suspicion of acute myocardial infarction (AMI). The final diagnosis was made according to the WHO criteria. The prevalence of AMI was 0.45 with a male to female ratio of 2:1. In all patients at least two blood samples were collected with 4 hours interval 4-12 hours after the onset of symptoms. All sera were analysed for myoglobin by a latex agglutination test and by a radioimmunoassay (RIA). The latex test was performed twice, first as an emergency test by the technical assistant on duty and later by another well-trained technical assistant as her daily routine work. If the latex test was carried out each day by the same well-trained technical assistant, the test results agreed well with the RIA test results, and the false-negative fraction for patients with AMI constituted 0.06 and the false-positive fraction for patients without AMI 0.46. However, when the latex test was performed by the occasional technical assistant on duty, a relatively high degree of discrepancy was observed between the latex test results and the RIA test results, thus giving a false-negative fraction of 0.11 and a false-positive one of 0.36. In conclusion, performed under optimal laboratory conditions, the latex test can be used as a reliable method to estimate elevated levels of serum myoglobin. However, used as a bedside emergency examination, the test results correlated rather poorly to the RIA test results, and consequently the latex myoglobin test seems to be of minor clinical importance in the early evaluation of patients with suspected AMI.

Female↗
PubMed1992

A latex agglutination test for the detection of Mycoplasma pneumoniae in respiratory exudates: a comparative study with a commercially available DNA-probe test.

We prepared polyclonal antibody specific to Mycoplasma pneumoniae. Using this antibody, we developed a latex agglutination test (LAT) for detecting the organism in respiratory exudates as rapid diagnosis of M. pneumoniae infection. Further, LAT was compared with DNA-probe test (DP) which was the only commercially available test for the rapid detection of the organism. In LAT, both M. pneumoniae and M. genitalium give positive agglutination, but the titer of M. genitalium was significantly lower than that of M. pneumoniae. The detection limit of LAT was 2 x 10(5) CFU/ml and that of DP was 5 x 10(4) CFU/ml in vitro. It was considered that target molecules in LAT were accumulated in the pharyngeal portion of the patients, because of their long half-life at 37 C. However, ribosomal RNA which was target molecule in DP was destroyed at 37 C much sooner, and the accumulation could not be expected. Actually, positive rate in LAT was higher than that in DP among clinical specimens in which M. pneumoniae was detected by culture method. The procedure of LAT is much easier and more rapid than that of DP in which radioactive isotope is required. LAT could be the choice of test for rapid diagnosis of M. pneumoniae infection.

Antigens, Bacterial↗
PubMed1975

Agglutinability of cattle red cells. 4. The effect of antiglobulin in comparison with treatment by pronase.

The average titre scores varied from zero to 24.7 for the cattle red cells (CRC) from different MZ pairs. These CRC were sensitized with different blood-typing reagents and were titre-tested against the double dilution series of an anti-bovine gamma-globulin serum. A significant negative correlation (r = - 0.82; P less than 0.001) was found between the degree of agglutinability and the amount of neuraminic acid of the surface component of CRC cleaved by pronase. After pronase treatment of the CRC it could be demonstrated that (1) the activity of the V and E'3 blood factors became destroyed; (2) three new specific receptors became evolved; (3) the degree of 'direct' agglutinability due to the A2, O3, W, S2 and Z anti-sera did not parallel with the titre scores obtained in the anti-globulin tests.

Absorption↗
PubMed1990

The use of D-Dimer assay by enzyme immunoassay and latex agglutination techniques in the diagnosis of deep vein thrombosis.

The blood of 107 patients undergoing venography for suspected deep vein thrombosis (DVT) was tested for D-Dimers using enzyme immunoassay (EIA) and two latex agglutination techniques. D-Dimer levels by EIA were raised in 88% of patients with proven DVT. However, levels detected with the two agglutination tests were much less sensitive, being raised in only 43% patients with DVT. A more sensitive rapid screening test is needed to provide a useful non-invasive test for the exclusion of DVT.

Adolescent↗
PubMed2004

Prostasome antigens as targets for sperm agglutinating antibodies demonstrated by 1-D gel electrophoresis and immunoblottings.

Many immunoinfertile men have sperm agglutinating antibodies that are directed against prostasome-derived antigens, but these antigens have not been defined so far. We selected serum samples with high ELISA titres against prostasomes from a group of immunoinfertile patients with sperm agglutinating antibodies and used the sera for immunoblottings on 1-D SDS-PAGE of prostasomes and sperm cells. The immunoblottings with individual antiprostasome antisera on 1-D SDS-PAGE of prostasomes, revealed three to 10 bands for each serum. Eighty-five per cent of the serum samples contained bands in the 70-75 kDa region and 80% of the samples contained bands in the 50-55 kDa region. Immunoblottings of extracted sperm cells, revealed one to six bands in the molecular weight range 25-82 kDa and two of the samples recognized two bands with molecular weights (50 and 43 kDa) similar to immunoblottings of prostasomes. The prostasomal antigens recognized by the high titre-antisera of immunoinfertile men were generally different from the sperm antigens recognized by the same sera. This suggests that prostasomes offer a new set of antigens available for research on male immunoinfertility and immunocontraception.

Antigens↗
PubMed2006

Development of non-agglutination microarray blood grouping.

Microarray technology provides an opportunity to monitor multiple parameters simultaneously. High-throughput applications such as blood donation screening could greatly benefit from performing various tests on a single testing platform. Blood grouping represents one part of the donation testing complementing the screening for blood-borne pathogens. Blood group serology traditionally exploited agglutination as the detection method. In this investigation, we have adapted blood grouping reactions to a solid-phase microarray substrate in a non-agglutination reaction format as an initial step in the development of a combined microarray testing platform. We have investigated immobilization of proprietary antibodies on multiple surfaces and monitored their performance under various reaction conditions. For the first time, highly specific blood grouping has been achieved on a planar microarray using directly labelled erythrocytes or a secondary labelled reagent using fluorescent signal end point readout. We have also complemented microarray data with a label-free, surface plasmon resonance-based Biacore platform data and used the real time quantitative measurement to rank anti-A antibodies according to the strength of reaction with the immobilized synthetic blood group antigen A.

Blood Grouping and Crossmatching↗
PubMed2004

Evaluation of a urinary antigen-based latex agglutination test in the diagnosis of kala-azar in eastern Nepal.

BACKGROUND: We evaluated the diagnostic accuracy as well as the reproducibility of the urine latex agglutination test 'KAtex' in the diagnosis of kala-azar in patients recruited at a tertiary care centre in Dharan, Nepal, between November 2000 and January 2002. METHODS: All patients presenting with fever of 2 weeks or more and splenomegaly were consecutively enrolled. Bone marrow and--if negative--spleen aspirates were examined for Leishmania donovani. Serum and urine samples were taken in duplicate for the Direct Agglutination Test (DAT) and KAtex. The reference laboratory determined sensitivity and specificity of KAtex. Reproducibility between both laboratories was assessed. RESULTS: KAtex was performed on urine from 155 parasitologically confirmed kala-azar and 77 non-kala-azar cases (parasitology and DAT-negative). KAtex showed a sensitivity of 47.7% (74/155, 95% CI: 39.7-55.9) and a specificity of 98.7% (76/77, 95% CI: 93.0-100.0). Reproducibility of KAtex showed a kappa of 0.684 (P < 0.001, n = 232). CONCLUSION: KAtex evaluation showed high specificity, low sensitivity and moderate reproducibility. A urine test for kala-azar could become a real breakthrough in kala-azar management if its reproducibility and sensitivity could be further improved.

Adult↗
PubMed1998

Use of the protective antigen of Erysipelothrix rhusiopathiae in the enzyme-linked immunosorbent assay and latex agglutination.

To establish a safe and convenient serodiagnostic method for swine erysipelas, a purified protective protein antigen of Erysipelothrix rhusiopathiae, which included a large amount of protective protein (64 kDa protein), was used for enzyme-linked immunosorbent assay (ELISA) and the latex agglutination (LA) test. In the ELISA, the antisera to four different serovars (1a, 2, 5 and 20) of E. rhusiopathiae exhibit a positive reaction, while antisera to other species of bacteria (Listeria monocytogenes, Staphylococcus aureus, Streptococcus suis, Rhodococcus equi and Corynebacterium pseudotuberculosis) exhibit a negative reaction. In the LA test, the antisera to three different serovars (1a, 2 and 5) of E. rhusiopathiae reacted with P64-sensitized latex beads, while the antiserum to serovar 20 (2553 strain) did not. Moreover, the antisera to other species of bacteria (Listeria monocytogenes, Staphylococcus aureus, Streptococcus suis, Rhodococcus equi and Corynebacterium pseudotuberculosis) did not in this test. Comparing the results of the growth agglutination (GA), ELISA and LA tests of 284 swine sera, there was a high degree of correlation among the results. The detection of anti-E. rhusiopathiae antibodies in the GA, ELISA and LA tests were compared using sera from pigs immunized with P64, alkaline extract (AE) and live-cell vaccine (LV). In all three tests, anti-E. rhusiopathiae antibodies could be detected 1 week after immunization. The serum antibody titre as determined by the LA test increased moderately, as did that by the GA test, while that determined by ELISA increased rapidly. These results suggested that ELISA could be used to monitor changes in anti-E. rhusiopathiae antibody titre and the LA test could be used in the screening test for swine erysipelas.

Animals↗
PubMed2006

A slide latex agglutination test for the rapid detection of antibodies in serum against porcine parvovirus.

A slide latex agglutination test (LAT) was developed and evaluated to detect serum antibodies against porcine parvovirus. Porcine parvovirus antigen was obtained by 10% PEG-6000 and 0.5 mol/l sodium chloride precipitation, and inactivated by 0.1% methanal. Two per cent suspensions of latex particles (0.5-0.8 microm) were coated by adding an equal volume of porcine parvovirus antigen at 0.34 microg/ml. Repeatability of latex agglutination test was evaluated with a panel of 100 sera using the same and different antigen lots. A good agreement between LAT and haemagglutination inhibit assay was observed. Because of convenience and speed of performance, this method would be used widely in clinic examination.

Animals↗
PubMed1998

Utility of a rapid fecal latex agglutination test detecting the neutrophil protein, lactoferrin, for diagnosing inflammatory causes of chronic diarrhea.

OBJECTIVE: The utility of tests for fecal neutrophils in the setting of chronic diarrhea has not been established. The purpose of this study was to determine the causes of chronic diarrhea associated with fecal neutrophils. METHODS: One fecal specimen from each of 10 normal subjects, 26 patients with known microscopic colitis, 13 with celiac sprue, eight with Crohn's disease, four with ulcerative colitis, and 103 with chronic diarrhea of unknown origin, as well as 10 fecal specimens from a patient with chronic nongranulomatous enterocolitis were analyzed blindly for the presence of a neutrophil granule protein called lactoferrin using a commercial latex agglutination kit. Diagnostic evaluation of the 103 patients with chronic diarrhea was carried out to determine the diagnostic accuracy of this test for chronic inflammatory bowel disease. RESULTS: None of the normal control subjects, three of 39 patients with microscopic colitis or celiac sprue, all 10 specimens from the patient with enterocolitis, and all 12 control patients with ulcerative colitis or Crohn's disease had a positive fecal lactoferrin test. Eleven of 103 patients with chronic diarrhea presenting without a diagnosis had a positive test, and all were diagnosed with an inflammatory condition of the colon (five-, ulcerative colitis; four-, Crohn's disease; one-, ischemic colitis; and one-, microscopic colitis). Only one patient with inflammatory bowel disease had a negative lactoferrin test. The sensitivity, specificity, and positive and negative predictive values of the fecal lactoferrin test for ulcerative or Crohn's colitis were 90%, 98%, 82%, and 99%, respectively. CONCLUSION: The major cause of fecal neutrophils in patients with chronic diarrhea is chronic inflammatory bowel disease of the colon. The latex agglutination test for fecal lactoferrin offers a highly sensitive, specific, and simple means for detection of fecal neutrophils in these patients.

Celiac Disease↗
PubMed1994

Evaluation of a New Latex Agglutination Test for Fecal Lactoferrin in Travelers' Diarrhea.

The detection and diagnosis of travelers' diarrhea by fecal culture method, often insensitive, expensive, and time consuming, could well be replaced with the latex agglutination test to detect fecal lactoferrin. As an initial screening, this test could be useful to differentiate between those patients who are more likely to have an invasive enteropathogen and those patients requiring antidiarrheal chemotherapy only. Fecal samples from 92 patients with travelers' diarrhea were tested for occult blood, fecal leukocytes, and fecal lactoferrin at stool dilutions of 1:50 and 1:200. The results were compared with findings from fecal cultures for enteropathogens. Statistical analyses were performed measuring the performance of the latex agglutination test for fecal lactoferrin to calculate its sensitivity, specificity, and positive and negative predictive values. Invasive pathogens were identified in 36 (39%), and a noninvasive pathogen was found in 18, or 20%, of the cases. At the stool dilution of 1:50, fecal lactoferrin showed more sensitivity than did leukocytes or occult blood in detecting the presence of invasive enteropathogens and, with a negative predictive value of 94%, was superior in predicting their absence. At the 1:200 dilution, a lower sensitivity of 55%, but a higher specificity of 82%, compared to 55% with the 1:50 dilution, was demonstrated. The determination of fecal lactoferrin might prove to be more useful and less expensive and time consuming as an initial screening for patients presenting with travelers' diarrhea. Further evaluation and antibacterial treatment could, therefore, be reserved for those with a positive lactoferrin test.

Journal Article↗