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PubMed1998

The latex agglutination test: standardization and comparison with direct agglutination and dot-ELISA in the diagnosis of visceral leishmaniasis in India.

Laboratory diagnosis of visceral leishmaniasis (VL) is usually based on the detection of Leishmania amastigotes in samples of bone marrow or splenic aspirate obtained by invasive procedures. Serological tests serve as a useful adjunct and are especially valuable in early or highly immune cases where amastigotes may be too scanty to be seen easily. The direct agglutination test (DAT) is generally considered the most suitable of the four types of tests currently employed (IFAT, counter immuno-electrophoresis, ELISA and DAT). However, the latex agglutination test (LAT) was recently reported to be a rapid and sensitive screening tool for VL and one which could be carried out at the patient's bedside. Further standardization and evaluation of LAT has now revealed that although it is comparable with DAT and dot-ELISA in terms of sensitivity it is far inferior because of cross-reactivity with other infections. This lack of specificity makes LAT unsuitable for routine diagnosis of VL even though it is rapid and sensitive. DAT still appears to be the best choice as a diagnostic tool, as it is very specific and does not require expensive equipment or reagents or much technical competence and the result can be visually interpreted. These merits make DAT very suitable for the diagnosis of VL in endemic areas of India.

Agglutination Tests↗
PubMed1986

Comparison of throat culture and latex agglutination test for streptococcal pharyngitis.

Numerous reports have recently appeared in the clinical microbiology literature that describe agglutination tests for identifying patients with group A streptococcal pharyngitis. These studies have indicated a close correlation between the results of the agglutination tests and traditional throat culturing. This paper describes a comparison study of 100 consecutive throat swab specimens using a commercially available agglutination test and routine throat culturing. The cultures were interpreted by an individual who was blinded to the agglutination test results. All agglutination testing was done by two laboratory members of a family practice office staff. The agglutination procedure was easy to perform and clear to interpret. The test sensitivity and specificity compared well with that reported in the literature from microbiology laboratories. The new agglutination tests are useful in the office laboratory for the identification of group A streptococci. Their primary advantage compared with throat culturing is the rapid availability of test results.

False Negative Reactions↗
PubMed1979

Serotyping of Serratia marcescens: simplified tube O-agglutination test and comparison with other serological procedures.

A simplified tube O-agglutination test was developed and evaluated for the determination of somatic serogroup (O) antigens of Serratia marcescens. Use was made of Tryptic Soy broth (TSB)-grown O-cells that had been boiled for 1 hour; 0.145 M NaCl proved a satisfactory diluent. Various technical parameters of this test were examined as well. Rabbit anti-O immune sera, that had been elicited with 5 x concentrated, TSB-grown, 1 hour-boiled O-cells of all 15 currently employed O-antigen reference strains of S. marcescens yielded satisfactory O-agglutinin titers. The tube O-agglutination test compared favorably with the indirect hemagglutination technic, although the latter technic yielded significantly higher O-agglutinin titers with merely 7 of the 15 O-antigens of S. marcescens. The tube O-agglutination test permitted detection of higher O-agglutinin titers than a microtiter O-agglutination test utilizing O-cells that had been stained with safranin O. Conversely, titers obtained with TTC-stained O-cells in a microtiter agglutination procedure approximated those yielded by the tube O-agglutination test, but O-cells of the various S. marcescens strains were stained nonuniformly by triphenyltetrazolium chloride (TTC). As before, there existed marked serologic cross-reactivity between O-antigens O6 and O14. A new O6 candidate strain, S. marcescens isolate S 1i, serotype O6:H20, was proposed. Contrary to O-agglutinins of human control serum, the O-agglutinins of rabbit anti-O immune sera proved refractory to treatment with 0.1 M of 2-mercaptoethanol (2-ME) and 0.01 M dithiothreitol (DTT) respectively. Dual absorptions of rabbit anti-O immune sera with killed cells of Staphylococcus aureus Cowan I (protein A), failed to significantly reduce O-agglutinin titers, although human IgG and IgM was bound by protein A. It was tentatively concluded that the 2-ME- and DTT-refractory rabbit anti-S. marcescens O-agglutinins resided in the IgM immunoglobulin class.

Agglutination Tests↗
PubMed2006

A meta-analysis of the diagnostic performance of the direct agglutination test and rK39 dipstick for visceral leishmaniasis.

OBJECTIVE: To compare the performance of the direct agglutination test and rK39 dipstick for the diagnosis of visceral leishmaniasis. DATA SOURCES: Medline, citation tracking, January 1986 to December 2004. Selection criteria Original studies evaluating the direct agglutination test or the rK39 dipstick with clinical visceral leishmaniasis as target condition; adequate reference classification; and absolute numbers of true positive, true negative, false positive, and false negative observations available or derivable from the data presented. RESULTS: 30 studies evaluating the direct agglutination test and 13 studies evaluating the rK39 dipstick met the inclusion criteria. The combined sensitivity estimates of the direct agglutination test and the rK39 dipstick were 94.8% (95% confidence interval 92.7% to 96.4%) and 93.9% (87.7% to 97.1%), respectively. Sensitivity seemed higher and more homogenous in the studies carried out in South Asia. Specificity estimates were influenced by the type of controls. In phase III studies carried out on patients with clinically suspected disease, the estimated specificity of the direct agglutination test was 85.9% (72.3% to 93.4%) and of the rK39 dipstick was 90.6% (66.8% to 97.9%). CONCLUSION: The diagnostic performance of the direct agglutination test and the rK39 dipstick for visceral leishmaniasis is good to excellent and seem comparable.

Agglutination Tests↗
PubMed1982

Detection of group B streptococcal antigen in early-onset and late-onset group B streptococcal disease with the Wellcogen Strep B latex agglutination test.

The Wellcogen Strep B latex agglutination test (Wellcome Diagnostics, Dartford, England) was evaluated as a method of detecting group B streptococcal antigen in urine, cerebrospinal fluid, and serum from neonates with early-onset (less than or equal to 7 days of age) and late-onset group B streptococcal disease. Urine was the best source of antigen, which was detected in 100% of six neonates with early-onset group B streptococcal disease who had urine available in the first 12 h of illness and in 88% of 17 group B streptococcus-infected neonates with urine available in the first 48 h of illness. Antigen was not detected in any samples from patients without group B streptococcal disease except in the urine of one patient with Proteus mirabilis meningitis. The Wellcogen Strep B latex test of the lot tested compares favorably with a noncommercially available latex agglutination test.

Antigens, Bacterial↗
PubMed1992

Latex agglutination and adenoviruses. I. Detection of polyclonal antibodies by latex agglutination test.

A latex agglutination (LA) test was developed for the detection and measurement of polyclonal anti-adenovirus antibodies. Latex particles, coated with purified hexon, penton or fibre antigens displayed specific agglutination with 21 rabbit immune sera directed against the different antigens and the complete virus of 10 human and 2 animal adenovirus types. The sensitivity of LA was compared to that of passive haemagglutination, complement fixation (CF) and immunodiffusion reactions. In experiments with immune sera as well as with human serum samples the sensitivity of LA test proved to be in the same range as that of CF test. Because of its easy and rapid performance (slide agglutination within 4 min) LA method may substitute for CF reaction in serological adenovirus diagnosis.

Adenoviridae↗
PubMed1992

Detection of occult blood in stools: comparison of three gaiac tests and a latex agglutination test.

The detection of occult blood in the stools is the only simple screening method for colorectal cancer. The aim of this study was to compare the results obtained with the new Hemolex kit (Orion diagnostica, Fumouze France) with those given by three gaiac tests--Hemoccult (Smithkline diagnostics), Hemofec (Boehringer Mannheim) and Hemopreuve (Fumouze) of 165 stools from patients without special diet. Seventy-one patients with at least two positive gaiac tests or a positive Hemolex test underwent colonoscopy followed, if negative, by fibroscopy: 28 had lesions of the lower digestive tract and five of the upper digestive tract. Sensitivity, specificity and negative and positive predictive value were of 70, 98, 91 and 92% respectively for Hemolex; 82, 74, 94 and 44% for Hemoccult; 94, 67, 98 and 42% for Hemopreuve and 91, 73, 97 and 46% for Hemofec. The results obtained in this study confirm the value of the Hemolex test for the detection of human occult blood in the stools whereas the gaiac tests used are influenced by dietary components (unless restricted), explaining their poor positive predictive value. In conclusion, due to their good negative predictive values, the authors recommend that screening for colorectal tumours should be based on the use of two or three gaiac tests which should be confirmed, when positive, by an immunological test for human hemoglobin.

Colonoscopy↗
PubMed1993

Rapid detection of human group C rotaviruses by reverse passive hemagglutination and latex agglutination tests using monoclonal antibodies.

Reverse passive hemagglutination (RPHA) tests and a latex agglutination test with monoclonal antibodies (MAbs) were developed for the rapid detection of noncultivatable human group C rotaviruses. For RPHA tests, two MAbs, MAb 5A12 recognizing the outer capsid and MAb 13A3 recognizing the inner capsid, were separately used for the coating of sheep erythrocytes (SRBCs). Forty-six fecal samples were examined to confirm the practicality of the tests. As a result, there was concordance between the RPHA test with SRBCs coated with MAb 5A12 and polyacrylamide gel electrophoresis of viral RNA (RNA-PAGE) in 44 (95.6%) of 46 samples, while the diagnoses by the RPHA test with SRBCs coated with MAb 13A3 were in complete agreement with those by RNA-PAGE. Furthermore, a latex agglutination test with MAb 13A3 was also developed, and this test was fast enough and sensitive enough to successfully detect the viruses from most fecal samples within 2 min. The present procedures would be useful for the diagnosis of human group C rotavirus infections in clinical laboratories which are not well equipped.

Antibodies, Monoclonal↗
PubMed2000

Evaluation of three newly developed enzyme-linked immunosorbent assays and two agglutination tests for detecting Salmonella enterica subsp. enterica serovar dublin infections in dairy cattle.

In this study test characteristics of three newly developed enzyme-linked immunosorbent assays (ELISAs) for Salmonella enterica subsp. enterica serovar Dublin were evaluated and compared with two agglutination tests. The ELISAs involved were an indirect ELISA with serovar Dublin lipopolysaccharide (LPS ELISA), an indirect ELISA with serovar Dublin flagellar antigen (GP ELISA), and a double-antibody sandwich blocking ELISA that uses monoclonal antibodies against S. enterica subsp. enterica serovar Enteritidis flagellin (GM-DAS ELISA). The agglutination tests involved were two routine serum agglutination tests with either somatic (O) or flagellar (H) antigen. Diagnostic specificity of the three ELISAs was determined using 840 serum samples from seven dairy herds without any history of serovar Dublin infection. Cutoff values at a titer of 100, 100, and 10, respectively, for the LPS ELISA, GP ELISA, and GM-DAS blocking ELISA resulted in a specificity of 99.3, 100, and 100%, respectively. Using these cutoff values the LPS ELISA, GP ELISA, and GM-DAS ELISA were able to detect, respectively, 30, 46, and 38% of 50 fecal culture-positive animals from 13 herds with a recent serovar Dublin infection. With the same cutoff values, active carriers (n = 18) were detected for 94.4% with the LPS ELISA and for 100% with the GP and GM-DAS ELISAs. Kappa values determined on the results of all tests from 8 of the 13 serovar Dublin-infected herds and the 7 control herds demonstrated a good correlation between the results of all ELISAs and the H-agglutination test. The results of the O-agglutination test failed to correlate with those of the other tests. Using a set of sera from 170 aborting cows (with 25 abortions due to serovar Dublin), test results of the ELISAs and the H-agglutination test were comparable. The H-agglutination test may be used successfully for single sample testing, especially to diagnose abortion due to serovar Dublin. It is concluded that the ELISAs are useful diagnostic tools in serovar Dublin control programs and that they are preferred to agglutination tests for reasons of automation and costs.

Abortion, Veterinary↗
PubMed1994

Relationship between direct agglutination test and splenic aspirate smear parasite load in visceral leishmaniasis at Baringo District, Kenya.

Direct agglutination test was carried out in Baringo District on 100 persons presenting with signs and symptoms suggestive of visceral leishmaniasis. Splenic aspirate smears and cultures were done on these 100 persons in order to parasitologically confirm the findings of the direct agglutination test. It was found that the direct agglutination test positively detected all 79 (79%) patients parasitologically confirmed to have visceral leishmaniasis. Irrespective of the splenic aspirate smear parasite rate, whether 1+ or 6+ on a logarithmic scale, direct agglutination test was positive. There were 21% false positives, two of whom had Schistosoma mansoni in their stools. It was not immediately known about the cause of the other false positives. It was concluded that the direct agglutination test is a good provisional serodiagnostic test for visceral leishmaniasis and should be considered for wider field application.

Adolescent↗
PubMed1985

Comparison of a latex agglutination test with five other methods for determining the presence of antibody against cytomegalovirus.

A latex agglutination test for determination of antibody against cytomegalovirus was compared with five other methods: a solid-phase fluorescent immunoassay, an indirect hemagglutination test, two solid-phase enzyme immunoassays, and an indirect fluorescent-antibody method, with sera collected from 210 random blood donors. Of the sera tested, 28% were positive for anti-cytomegalovirus by concordance of four or more methods. The latex agglutination test performed well, with a sensitivity of 100%, a specificity of 99%, and positive and negative predictive values of 97 and 100%, respectively. The methods were also evaluated for the number of sera requiring repeat testing, equivocal results after retesting, ease of performance, turnaround time, and technical demands. The tests which best met the requirements for a screening test were the solid-phase fluorescent immunoassay, the indirect hemagglutination test, and the latex agglutination test. The latex agglutination test is a valuable screening tool for detecting total anti-cytomegalovirus which has high sensitivity, high negative predictive value, and rare equivocal results and also has the added advantages of ease of performance and rapid turnaround time.

Antibodies, Viral↗
PubMed1996

Spot agglutination test for the rapid diagnosis of goat pox.

A spot agglutination test was standardised for the rapid diagnosis of goat pox using antigen and antibody systems after being coupled to glutaraldehyde-treated sheep erythrocytes. The sensitivity was comparable to that of counter-immunoelectrophoresis and reverse-phase passive haemagglutination tests. The spot agglutination test was found to be very simple, easy to perform, rapid and economical for the detection of goat pox antigen and antibody and can be performed under field conditions.

Agglutination Tests↗
PubMed1988

A rabies agglutination test (RAT) for rabies antibody detection.

An agglutination test has been developed for the detection of rabies antibodies after human vaccination. The rabies agglutination test (RAT) is based on the capability of specific antibody to agglutinate sensitized polystyrene (or latex) beads. In the RAT, latex beads were coated, in a first step, with inactivated and purified rabies virus (PV strain adapted and propagated on BHK-21 cells) and, in a second step, with bovine serum albumin. Negative control beads were coated with bovine serum albumin (BSA) only. To test for human antibody, several microliters of serum were mixed on a glass slide with an equal volume of virus-sensitized beads and the mixture was gently agitated. After a few minutes, agglutination was visible with sera which had been characterized as positive by the virus neutralization antibody (VNAb) technique. No agglutination was observed with negative sera tested with virus-coated beads or with positive sera tested with BSA-coated beads. Virus-sensitized beads were agglutinated when the virus neutralizing antibody titres were equal to or greater than 2.5 international units per ml (IU/ml) in human sera. The concordance between the RAT results and VNAb titres was about 97% when 2.5 IU/ml was taken as the cut off value for determining the positive sera with the VNAb technique. The possibility that clinicians might use the RAT as a simple means to determine sero-conversion at the end of the post-exposure treatment of patients is discussed.

Agglutination Tests↗
PubMed1973

Salmonella dublin abortion in cattle. II. Observations on the whey agglutination test and the milk ring test.

The serum and whey agglutination test were compared on paired samples from thirty-five cases of bovine abortion associated with Salmonella dublin infection. The whey test proved nearly as useful as the serum test for confirming an active infection though it was only practicable to examine the whey for flagellar antibodies. S. dublin was isolated from nearly half of the milk samples obtained within the first week of abortion but none of those collected after the fourth week. The whey test proved of no value in retrospective identification of abortion cases. The trial using the milk ring test was disappointing.

Abortion, Veterinary↗
PubMed2006

Diagnosing human African trypanosomiasis in Angola using a card agglutination test: observational study of active and passive case finding strategies.

OBJECTIVE: To assess the operational feasibility of detecting human African trypanosomiasis by active and passive case finding using the card agglutination test with serial dilution of serum to guide treatment. SETTING: Trypanosomiasis control programme in the Negage focus, northern Angola, during a period of civil war. DESIGN: Observational study. PARTICIPANTS: 359 patients presenting themselves to health centres with symptoms (passive case finding) and 14,446 people actively screened in villages. MAIN OUTCOME MEASURES: Whole blood and serological tests at different dilutions using the card agglutination test, and detection of parasites by microscopy. RESULTS: Active case finding identified 251 people with a positive card agglutination test result, 10 of whom had confirmed parasites. In those presenting for investigation 34 of 51 with a positive card agglutination test result at the dilution of 1:8 or more used to guide treatment had parasites in blood, lymph node fluid, or cerebrospinal fluid, compared with 10 of 76 in those detected by active case finding: positive predictive values of 67% for passive case detection and 13% for active case detection. Only at a cut-off dilution more than 1:32 was the positive predictive value in active case detection reasonable (46%) and at this dilution 40% of microscopically proved cases were missed. CONCLUSIONS: The card agglutination test is useful for initial screening in active detection of cases with human African trypanosomiasis but, given the toxicity of the drugs, serology using the card agglutination test should be not used alone to guide treatment after active case finding. A second confirmatory test is needed.

Adolescent↗