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PubMedDate not supplied

Serodiagnosis of melioidosis by a competitive enzyme-linked immunosorbent assay using a lipopolysaccharide-specific monoclonal antibody.

Burkholderia pseudomallei is the causative agent of melioidosis, a severe and potentially fatal infectious disease in humans known to be endemic in Southeast Asia and northern Australia. The infection is also increasingly recognized in various animal species with a potential to spread to humans. With the potential as a biological warfare agent, specific serodiagnosis of melioidosis for surveillance in large populations at risk, humans or animals, would be highly valuable. In this study, a competitive enzyme-linked immunosorbent assay (ELISA) using a lipopolysaccharide-specific monoclonal antibody was developed. The assay provides high specificity, based on a previously described monoclonal antibody to a specific epitope on the lipopolysaccharide (LPS) of B. pseudomallei. The assay sensitivity of 96.0% and specificity of 100% were achieved at a cutoff value of 50% inhibition in human culture-proven melioidosis cases. An optimal cutoff value of 65% inhibition for sera from a melioidosis endemic area was obtained by ROC analysis and resulted in an assay specificity of 86.2%, while maintaining assay sensitivity of 92.0%. A potential application of the assay in the serodiagnosis of melioidosis in animal species was also evaluated usina dolphin sera with satisfactory results.

Animals↗

PubMed1991

Serodiagnosis of cancer by using Candida cytochrome c recognized by human monoclonal antibody HB4C5.

Cytochrome c from various sources, such as Candida krusei, yeast, horse, and cattle, was found to be recognized by human monoclonal antibody HB4C5 specific to lung cancer. Therefore, the cytochrome c was applied to the measurement of antibody amount in patient sera with a similar reactivity to the antibody HB4C5 for serodiagnosis of cancer. The cytochrome c from Candida krusei was most valuable for the serodiagnosis of various cancers, and the yeast cytochrome c was also useful. However, horse and bovine cytochrome c did not react with antibody of the cancer patients. By using Candida cytochrome c lung, bile duct, esophagus, and liver cancers were detected at high rates of more than 50%. In the case of lung cancer, the detection rates of small-cell, squamous, large-cell and adenocarcinoma were 78%, 63%, 100%, and 34%, respectively. The rate for small-cell carcinoma was higher than that with the currently used NSE assay system, and the rate for squamous carcinoma was comparable to that with the SCC assay system, although the system using cytochrome c did not show similar reactivity to that with the SCC system. Furthermore, lung cancer was detected at early stages by using cytochrome c, and even in the case of adenocarcinoma, the rate at early stages with the cytochrome c system was higher than that with the CEA assay system. On the other hand, false positive rates of benign diseases and normal were low--8% and 2%, respectively.

Antibodies, Monoclonal↗

PubMed2006

Western immunoblot analysis using a ten leptospira serovars combined antigen for serodiagnosis of leptospirosis.

Leptospirosis is a major zoonotic disease throughout the world. There are unavailable accuracy diagnostic methods for the acute phase of infection. To demonstrate the advantage of Western immunoblot, a mixed leptospira serovars antigen for the serodiagnosis of leptospirosis was employed. SDS-PAGE and Western immunoblot was performed using a 10 mixed leptospira serovars antigen and stained with 16 reference rabbit anti-leptospirosis antibodies. The result showed different immunoreactive band patterns for each reference serum. The bands with molecular weights of 15-20, 23-24, 41 and 45 kDa were commonly found (88% to 100% of the 16 reference sera). Using combined leptospira antigens in a Western immunoblot technique is an alternative and practical strategy for a more sensitive leptospirosis serodiagnosis.

Antibodies, Bacterial↗

PubMed1991

Serodiagnosis of melioidosis in Singapore by the indirect haemagglutination test.

Melioidosis is endemic in Singapore, with diagnosis dependent upon both bacteriological culture and serodiagnosis. Using the polysaccharide (melioidin)-sensitized turkey red cells in the indirect haemagglutination test (IHAT), 20 (100%) of the Pseudomonas pseudomallei culture-positive cases were detectable by the IHAT with titles ranging from 1:16 to 1:32, 768. Eight of these patients who died within a few days after the IHAT was performed had titres ranging from 1:16 to 1:1028. Five culture-negative patients, with clinical symptoms suggestive of melioidosis infection and who responded to treatment with ceftazidime, showed IHA titres between 1:64 and 1:8,192. One hundred and twenty one sera from patients with pneumonia, abscesses, or diabetes mellitus were IHAT negative. The IHAT showed good specificity since negative titres were seen in tests using sera from 2 patients with culture-positive Pseudomonas aeruginosa and 4 patients positive for Legionella. IHAT negative results were obtained from tests of 50 normal blood donors and 50 sewerage workers. Of 683 national servicemen tested, 5 (0.73%) had IHAT titres ranging from 1:16 to 1:128. Unlike hyperendemic areas such as Thailand where interpretation of IHAT is seriously hampered by IHA titres found in one-third to half of the population, serodiagnosis of melioidosis by the sensitive IHAT may be employed in Singapore as a routine procedure since background IHA titres are low.

Hemagglutination Tests↗

PubMed1991

Dot enzyme immunosorbent assay for the serodiagnosis of typhoid fever.

A nitrocellulose membrane strip dotted with a specific 50 kDa outer membrane protein of Salmonella typhi was applied for the serodiagnosis of typhoid fever. Using horseradish peroxidase conjugated IgM and IgG antibodies with 4-chloronaphthol as substrate, antibodies in typhoid patients were clearly visualised as bluish purple dots while sera from patients with non-typhoid fevers gave negative results. The detection of specific IgM and IgG antibodies in typhoid patients suggest either recent or current infection. Combined with the high specificity, reliability and rapidity of the test, the dot EIA technique provides a simple and useful method for the serodiagnosis of typhoid using a single serum specimen.

Enzyme-Linked Immunosorbent Assay↗

PubMed1990

Multi-dot enzyme-linked immunosorbent assay for serodiagnosis of trematodiasis.

A nitrocellulose membrane strip dotted with crude antigens of trematodes, such as Paragonimus miyazakii, P. westermani, Fasciola sp., Clonorchis sinensis and Schistosoma japonicum was applied for serodiagnosis of trematodiasis. With peroxidase conjugated anti-human IgG and 4-chloro-1-naphthol as a substrate, antibodies in trematodiasis patient sera were clearly detected on the strip as bluish purple spots. Densities of the spots correlated well with values obtained by standard micro-ELISA. Cross-reactions among the trematode antigens, which is occurred more frequently than by ELISA, were observed with this method. However, the darkest spot correlated with the infecting parasite in all cases. With the high reliability and advantage that crude antigens can be used, this simple method, multi-dot ELISA, is considered to be useful for serodiagnosis of trematodiasis.

Animals↗

PubMed1989

Efficacy of immunoprecipitation methods for rapid serodiagnosis of allergic bronchopulmonary aspergillosis.

The relative efficacy of Ouchterlony's double diffusion (ODD), Wadsworth's gel diffusion (WGD) and counter-immunoelectrophoresis (CIE) is reported for serodiagnosis of allergic bronchopulmonary aspergillosis (ABPA). Of the 50 sera taken from patients with confirmed or suspected ABPA, serum precipitins were demonstrated against Aspergillus fumigatus by CIE, WGD and ODD in sera from 34, 31 and 25 patients, respectively. The results were negative by the three methods in sera taken from 10 healthy volunteers. It is concluded that CIE followed by WGD is more efficacious than ODD for serodiagnosis of ABPA.

Antibodies, Fungal↗

PubMed1989

Rapid indirect haemagglutination (rapid-IHA) using sensitized chick cells for serodiagnosis of amoebiasis at primary health centre level.

A successful modification of the indirect haemagglutination (IHA) test as a simple and rapid diagnostic procedure for serodiagnosis of amoebiasis at a primary health centre (PHC) level laboratory has been described in the present study. In this modified IHA test, the chick red blood cells (RBC), stabilized by the double aldehyde method, could be sensitized with amoebic antigen and then stored at 4 degrees C for up to 45 days for their subsequent use, as ready-made reagent directly in the IHA, without loss of sensitivity. The chick red cells settled quickly and their haemagglutination pattern could be determined within 30 to 45 min of incubation at room temperature with test sera. The IHA using double-aldehyde stabilized (DAS) chick cells, showed equal sensitivity and specificity with that of IHA using human O cells in diagnosing cases with different manifestations of amoebiasis. With the use of stored sensitized DAS chick cells directly in the IHA, the test could be performed and results obtained within 60 to 90 min of receipt of the sera to be tested. This makes IHA a simple and rapid procedure for its adaptation at a PHC level laboratory for the serodiagnosis of amoebiasis.

Amebiasis↗

PubMed1988

[Serodiagnosis of toxoplasmosis developing during pregnancy and of congenital toxoplasmosis].

During the first prenatal serodiagnosis of toxoplasmosis, the test must permit to differentiate between immunized and non-immunized patients and to screen recently contracted toxoplasmosis. In a group of 33 women affected with toxoplasmosis during pregnancy a critical study of serodiagnosis criteria is carried out by comparing the theoretical protocol of the evolution of the serology during acquired toxoplasmosis with the situations observed under usual prenatal monitoring. Seroconversion was noted in 26 women and the variability of the results emphasizes the difficulties in determining the date of the contamination when an evolutive form of toxoplasmosis is suspected at the first examination, which is the case in 7 other patients. In children, 11 congenital toxoplasmosis were diagnosed, all on laboratory examinations. It must be emphasized that 16 children were prematurely lost to follow-up. It should be necessary to devote our energies to screening and information in order to validate the protocol of prevention of congenital toxoplasmosis.

Agglutination Tests↗

PubMed1986

Serodiagnosis of histoplasmosis.

The standard serologic tests for diagnosis of histoplasmosis include the complement fixation test (CF), which is quite sensitive, and the immunodiffusion test (ID), which is fairly specific. Radioimmunoassay (RIA) is an experimental serological test even more sensitive than CF, but also less specific. Problems with the specificity of serodiagnosis are addressed followed by a discussion of the sensitivity of the three tests in the different forms of histoplasmosis-chronic cavitary histoplasmosis, progressive disseminated histoplasmosis, and acute pulmonary histoplasmosis. Next, serologic results from a large point-source outbreak of acute pulmonary histoplasmosis in Orono, Minn, are presented. These results are used to define the sensitivity of the three tests in acute symptomatic pulmonary histoplasmosis and also to detail the time course of the appearance of antibodies measurable by each test. Finally, a case of sporadic severe acute pulmonary histoplasmosis is presented in which the approach to serodiagnosis was based on lessons learned from the Orono outbreak.

Adult↗

PubMed1985

Comparative study of four saprophytic leptospira strains as screening antigens in the serodiagnosis of leptospirosis in water buffaloes (Bubalus bubalis).

The profitability of four saprophytic leptospira strains (Buenos Aires, Patoc 1, Rufino and São Paulo) as polyvalent antigen in the serodiagnosis of leptospirosis in water buffaloes was studied by the microscopic agglutination test. From 104 examined sera tested against 16 pathogenic leptospira serotypes, 52 were positives and 52 were negatives. The results led to the conclusion that from four studied strains, Buenos Aires strain showed the best results when utilized in screening tests for the serodiagnosis of leptospirosis in water buffaloes. The other three strains showed a good specificity but their sensibilities were poor and therefore should not be recommended for diagnosing water buffalo leptospirosis.

Animals↗

PubMed1968

Water strains of Leptospira in the serodiagnosis of human and animal leptospirosis.

The most widely used serological reaction for the diagnosis of leptospirosis is the agglutination test. This test, however, cannot be carried out in many laboratories because special equipment and special experience are required. It is also necessary to maintain live Leptospira cultures belonging to all the serotypes present in the country where the test is made. Consequently, it would be extremely useful to be able to diagnose leptospirosis by means of a single antigen, regardless of the serotype to which the Leptospira responsible for the infection belonged. This is particularly important for countries in which the antigenic pattern of the local leptospirae is not well known and in which it would thus be necessary to use a large number of Leptospira serotypes for each test.Observations made in the last 11 years have suggested that the problem may be solved with the use of some non-pathogenic, water strains of Leptospira which seem to be agglutinated by human sera containing antibodies against pathogenic leptospirae. This paper reports the results of studies from 1960 to 1968 on the possibility of using water strains for serodiagnosis.The results over the 8-year period show that the non-pathogenic strain Patoc 1 is agglutinated by a high percentage of human sera positive for pathogenic leptospirae: these results indicate that Patoc 1 would be useful for serodiagnosis. However, a high percentage of animal sera positive for pathogenic leptospirae gave negative results with the strains Patoc 1 and Sao Paulo, and thus these strains cannot be used for serodiagnosis in animals.

Agglutination Tests↗

PubMed1984

Examinations on the serodiagnosis of Sarcocystis infections. II. Class-specific immunoglobulin responses in mice, pigs, and sheep.

Mice, pigs and sheep were experimentally infected with Sarcocystis muris, S. miescheriana (syn. S. suicanis) and S. tenella (syn. S. ovicanis) respectively. Enzyme-linked immunosorbent assays employing conjugates specific for the relevant host immunoglobulin classes M and G were then evaluated for use in serodiagnosing these infections. Soluble antigenic extracts were prepared from cystozoites harvested from macroscopic S. muris and S. gigantea (syn. S. ovifelis) cysts and from microscopic S. miescheriana cysts. Selected sera were firstly screened in the immunoassays for false positive and negative reactions which may have resulted from competition or interference occurring between the different antibody classes. No false reactions were detected as evidenced by the reactions of the sera before and after mercaptoethanol reduction of IgM and gel fractionation of IgM and IgG. The attempted absorption of IgG from the sera with Protein A proved ineffective in all 3 host animal species. Sera from the experimentally infected animals were then tested in the immunoassays against homologous and heterologous antigens and the kinetics of specific antibody development were determined. None of the antigens proved to be species-specific although heterologous reactions were usually weaker. Specific IgM-antibodies were detected early in the courses of all infections but they did not persist. These transitory responses occurred simultaneously with parasite merogonous proliferation in mice and sheep but later in pigs. Such responses may be of some value in the serodiagnosis of acute sarcocystosis. Specific IgG-antibodies were detected later in all infected animals and elevated levels were prominent after cyst formation by the parasite had occurred. These responses were therefore suitable for the serodiagnosis of latent and chronic Sarcocystis infections.

Animals↗

PubMed1981

Fluorescent leprosy antibody absorption (FLA-ABS) test for early serodiagnosis of leprosy.

The conventional parameters for the diagnosis of leprosy i.e. clinical symptoms aided by smear and histological examinations, have certain limitations. With the conventional parameters, it is difficult to diagnose new cases in the early stage of leprosy. Recently Abe et al (1976) and Abe and Yoshino (1978) suggested that Florescent Leprosy Antibody Absorption (FLA-ABS) test developed by one of the authors (Abe) might be useful for early serodiagnosis of leprosy. Following this, the FLA-ABS test was standardised at the Central Jalma Institute for Leprosy, Agra to study the usefulness of the test in early serodiagnosis and detection of sub-clinical infections. In the present study sera were collected from 136 cases of different types of leprosy which includes 11 cases with lesions suspected to be leprosy, 58 healthy contacts and 19 healthy controls. Using FLA-ABS test, anti M. leprae antibodies were detected in all types of leprosy irrespective of the type and duration of the disease. The test has shown significantly high positive results in very early cases of leprosy with a duration of 3-6 months and having very small lesions measuring below 2 cms.

Fluorescent Antibody Technique↗

PubMed1994

[Serodiagnosis of caseous lymphadenitis (pseudotuberculosis) in sheep].

Random postmortal isolation of Corynebacterium pseudotuberculosis from the sheep used in a research project and the consequently determined serological positivity of these animals indicated the serological examination of the flock from which the sheep came. No symptoms of caseous lymphadenitis were ever observed in the flock in question and no data on the occurrence of this disease were available in the flock history. The sera of 228 head of sheep were used for examinations, while this number of sheep represented half the flock head. Agar-gel immunodiffusion (AID) and neutralization of the toxin C. pseudotuberculosis (NTX) were applied as serological assays. The antigen in both tests was an isolated form of toxin i.e. phospholipase D (PLD). To determine the effect of PLD, the intensity of its synergic hemolytic effect with the equi factor Rhodococcus equi was found out and it was expressed in activity units (AU). Undiluted serum with PLD containing 5,000 AU/ml was examined in AID. Positive precipitation was observed in 79 examined sera, out of which 71 were also positive in NTX. NTX demonstrated the inhibition of synergic hemolytic activity of PLD with the equi factor, which was used to sensibilize sheep erythrocytes in agar medium. PLD with the terminal content of 10 AU/ml was used, as well as twofold dilution series of sera. The first evaluated dilution series was 1:10. In this dilution series, the highest number of sera, 22, had the positive reaction. The highest positivity at a dilution series 1:5,120 was found out in one serum. A total of 71 sera had positive reactions in NTX and those sera were also positive in AID. The determined 34.7% positivity substantiated the importance of serodiagnosis for the intravital diagnosis of caseous lymphadenitis of sheep, particularly of its chronic form. AID and NTX are usable tests for serodiagnosis of this disease.

Animals↗

PubMed1993

Evaluation of the potential role of enzyme-linked immunosorbent assay in serodiagnosis of tuberculosis.

The serodiagnosis of tuberculosis was evaluated by means of enzyme-linked immunosorbent assay of IgG antibody to mycobacterial antigen 60. The study population consisted of 63 patients with active tuberculosis comprising 43 pulmonary tuberculosis, 15 tuberculosis pleurisy, four miliary tuberculosis, and one tuberculous lymphadenitis. The control group consisted of 48 patients embracing 8 cases of inactive pulmonary tuberculosis, 31 cases of pulmonary disorders other than tuberculosis, and nine normal persons. There was a significant difference in antibody titers of sera between patients of active tuberculosis and control group (P < 0.001). The sensitivity of the test for active pulmonary tuberculosis was 81.4% and the specificity was 85.4% (cutoff value 300 units). The sensitivity for all active tuberculosis was 61.9%. There was positive correlation between serum titer of active pulmonary tuberculosis and the extent of disease on chest radiography (r = 0.4, P < 0.01). A positive correlation was found between the titers of pleural fluids, and the corresponding sera in 15 patients (eleven tuberculous pleurisy and four nontuberculous controls) (r = 0.85, P < 0.001). Whether serum or pleural fluid, the positive rate of the assay for tuberculous pleurisy was relatively low. In conclusion, the A60 IgG ELISA test may be useful in serodiagnosis of pulmonary tuberculosis and assessment of the disease extent, although it might not yield a high sensitivity for tuberculous pleurisy.

Antibodies, Bacterial↗

PubMed1996

[Serodiagnosis of toxoplasmosis: a comparative multicenter study of a standard scale through various actual tests and expression of the results in international units. Groupe de travail toxoplasmose du Contrôle national de qualité en parasotologie. Syndicat des fabricants de réactifs de laboratoire. Groupe de travail standardisation des tests sérologiques du Réseau européen de lutte contre la toxoplasmose congénitale].

Reported are the results of a multicentre study involving 40 laboratories that was carried out in France to assess all the currently available methods used for the serodiagnosis of toxoplasmosis. For this purpose 10 batches of control sera were prepared with titres in the range 0-260 IU per ml. These sera were tested in nine laboratories using immunofluorescence methods; in three laboratories using dye tests; in forty laboratories using enzyme-linked immunosorbent assay; in four laboratories using direct agglutination and haemagglutination; in seven laboratories using the high-sensitivity IgG agglutination test; and in three laboratories using the latex agglutination test. In this way, 70 series of titrations were carried out using seven procedures and the results were compared with those obtained using the WHO reference serum in 15 cases, with the French national E6 serum in 16 other cases, and in 39 cases using 15 reference sera supplied by the reagent manufacturers. Rigorous comparison of the tests was not possible in all cases because one aim of the study was to ensure that the tests were carried out under the usual working conditions that prevailed in the participating laboratories. The results obtained indicate that the serological tests currently available for toxoplasmosis are acceptable for its serodiagnosis. Presentation of the titres in IU has advantages; however, caution is required since the definition of IU varies according to the test and reagents used. It is therefore essential that the conditions and limits for a positive reaction be carefully defined in each case, especially for commercially available kits.

Agglutination Tests↗

PubMed1996

Comparison of adult somatic and excretory-secretory antigens in enzyme-linked immunosorbent assay for serodiagnosis of human infection with Fasciola gigantica.

Adult somatic antigen extract of Fasciola gigantica was compared with excretory-secretory (ES) antigen in an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of human fascioliasis gigantica. The absorbance values in ELISA using the adult somatic antigen were not significantly different from the values obtaining using ES antigen (p > 0.05). The diagnostic sensitivity, specificity and positive and negative predictive values of the test using adult somatic extract as antigen were 100%, 98%, 70% and 100%, respectively. On the other hand, these values of the test using adult ES antigen were 100%, 99.3%, 87.5% and 100%, respectively. It appears that both somatic and ES antigens are effective antigens for use in the serodiagnosis of human fascioliasis gigantica.

Animals↗