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Detection of parainfluenza IgM antibody by hemadsorption immunosorbent technique.

A hemadsorption immunosorbent technique (HIT) was developed for the detection of immunoglobulin M (IgM) to parainfluenza virus types 1, 2, and 3. Twenty-six (90%) of twenty-nine patients under 6 yr of age from whom parainfluenza virus was isolated showed parainfluenza IgM antibody in one or both of their paired sera, with titres ranging from 320 to 81,920. In about one third of the cases IgM antibody was demonstrated in the initial sera taken 1 to 3 days after the onset of illness. Heterotypic IgM antibody responses were observed in about 20% of the patients. The HIT test was more sensitive than the hemagglutination inhibition (HI) and complement fixation tests in detecting a seroresponse in the 29 virus-positive children. The results of studies in older patients with HI titre rises to parainfluenza virus suggested that reinfection probably induced IgM antibody in a proportion of cases. The HIT test proved to be specific for the IgM class of antibody and avoided false-positive results due to rheumatoid factor. It permits an early presumptive diagnosis in a proportion of patients with parainfluenza infection.

Adolescent↗

Hemadsorption immunosorbent technique for determination of mumps immunoglobulin M antibody.

We used hemadsorption immunosorbent technique (HIT) to detect mumps immunoglobulin M (IgM) antibody. IgM from human sera was adsorbed into anti-human IgM-coated wells in plates, and mumps-specific IgM was detected by adding mumps virus hemagglutinin and guinea pig erythrocytes consecutively. Specific IgM-positive sera showed hemadsorption, whereas negative sera showed hemagglutination. All 81 patients with current mumps infections tested showed mumps-specific IgM antibody, with titers ranging from 160 to 327,680. In most cases IgM antibody was already present on day 1 or 2 after the onset of illness. IgM antibody persisted for 6 to 10 weeks. Of 57 patients with acute respiratory illnesses caused by parainfluenza virus, 5 showed cross-reactions in the hemadsorption immunosorbent technique assay for mumps IgM. The hemadsorption immunosorbent technique assay is specific for the IgM class of antibody and avoids false-positive results due to rheumatoid factor. This test is an efficient and sensitive method for rapid and early diagnosis of mumps infections.

Animals↗

Hemadsorption immunosorbent technique for determination of rubella immunoglobulin M antibody.

A highly specific and sensitive hemadsorption immunosorbent technique for measuring rubella immunoglobulin M (IgM) antibody is described. IgM from human sera was absorbed into anti-human IgM-coated wells in plates and rubella-specific IgM was detected by adding rubella virus hemagglutinin and a small quantity of sheep erythrocytes. Centrifugation of the plates facilitated reading of the test. Specific IgM-positive sera showed hemadsorption, whereas negative sera showed hemagglutination. Rheumatoid factor and rubella-specific IgG antibody did not interfere with the results. The test was clearly more sensitive than the solid-phase immunosorbent technique for detection of rubella IgM antibody by hemagglutination inhibition and at least as sensitive as the hemagglutination inhibition test on IgM fractions from a sucrose density gradient and the indirect immunofluorescence test for IgM antibody with absorbed serum. All of 40 sera from 17 rubella patients taken 4 to 49 days after the onset of rash were positive in the new test, with antibody titers ranging from 2,560 to 81,920 between 4 and 28 days. The test is reliable, practical, and suitable for general diagnostic use.

Antibodies, Viral↗

Hemadsorption immunosorbent technique for the detection of dengue immunoglobulin M antibody.

We developed a highly specific, sensitive, and economical hemadsorption immunosorbent technique for the detection of dengue-specific immunoglobulin M (IgM) antibody. The technique is based on the reaction of human sera with anti-human IgM immobilized onto a solid phase followed by the detection of dengue-specific IgM by the addition of a known quantity of dengue virus hemagglutinin and goose erythrocytes. Dengue-specific IgM-positive sera showed hemadsorption. IgM antibody specific for dengue virus was detected in 22 of 39 (56%) convalescent-phase sera from primary dengue infections and 8 of 10 (80%) convalescent-phase sera from secondary dengue infections. Additionally, 32 of 76 single sera from patients were positive for dengue IgM; these sera were previously uninterpretable by the hemagglutination inhibition test, as only a single serum specimen was available. No false-positive results were obtained with sera that were negative by the hemagglutination inhibition test for dengue virus. Crude dengue virus hemagglutinin preparations could be used without purification. Dengue-specific IgG did not interfere with the results, nor was there any cross-reactivity between dengue hemagglutinins and IgM specific for other viruses. Some cross-reactivity of the dengue-specific IgM was observed with Japanese encephalitis virus hemagglutinins, but this did not present any problems in the interpretation of results. This test is specific, inexpensive, highly reproducible, and simple to perform.

Antibodies, Viral↗

Solid-phase immunosorbent technique for rapid detection of Rift Valley fever virus immunoglobulin M by hemagglutination inhibition.

A solid-phase immunosorbent technique (SPIT) was adapted to detect Rift Valley fever (RVF) virus-specific immunoglobulin M (IgM) in serum samples from humans vaccinated with Formalin-inactivated RVF vaccine. Microdilution plates coated with goat anti-human IgM were successively incubated with serum samples from human vaccinees, RVF virus hemagglutinating antigen, and goose erythrocytes. The RVF virus-specific IgM in the serum samples from vaccinees bound to the RVF virus antigen and inhibited hemagglutination of goose erythrocytes. SPIT was compared to the IgM capture enzyme linked immunosorbent assay (ELISA) and the indirect immunofluorescent-antibody (IFA) assay and was found to be sensitive in detecting RVF virus-specific IgM antibody, with high correlations between SPIT and the other two tests (Pearson's correlation coefficient [r] = 0.9 and 0.6, respectively). Results of SPIT were obtained within 5 h, offering speed over ELISA (8 h). In addition, SPIT does not require sophisticated equipment or expensive reagents. Serum rheumatoid factor did not produce false-positive reactions in SPIT as in the indirect immunofluorescent-antibody assay and IgM capture ELISA.

Antibodies, Viral↗

Preliminary studies on the use of solid-phase immunosorbent techniques for the rapid detection of Wesselsbron virus (WSLV) IgM by haemagglutination-inhibition.

Serum samples from 446 randomly selected persons belonging to different age groups and locations in Nigeria were tested for the presence of WSLV IgM using the flavivirus haemagglutination-inhibition (HI) test adopted to the solid-phase immunosorbent technique (SPIT). 61 (14%) persons had IgM to WSLV only, while 9 (2%) persons had heterologous IgM to WSLV and two other flaviviruses, namely yellow fever and Uganda S viruses. There was a high prevalence of IgM in people of younger age groups than those in older groups. The majority of the IgM positive sera (67 (96%) of the 70 positive sera reacted to high titres (>21:80). With the conventional HI tests, 314 (70%) of the total sera tested had HI antibodies to one or more flaviviruses (yellow fever, West Nile, Potiskum, Zika and Uganda S) out of which 305/314 (97%) had antibodies to 3 or more flaviviruses used in the tests. Although SPIT may not be as sensitive as the conventional HI test, it was found to be more specific and could be adopted for the detection of early WSLV infections in flavivirus hyperendemic environments.

Adolescent↗

A solid-phase immunosorbent technique for the rapid detection of rubella IgM by haemagglutination inhibition.

The rubella haemagglutination inhibition (HAI) test has been modified for the detection of rubella-specific IgM. The rubella HAI test is performed in microtitre plate wells in which IgM from patients' sera has been selectively retained by anti-human IgM bound to the polystyrene surface. The test requires only anti-human IgM-coated microtitre plates, in addition to the standard rubella HAI reagents. The results obtained in this test, a solid-phase immunosorbent technique (SPIT), are in good agreement with results obtained by the sucrose density gradient centrifugation technique (DGCT). Advantages include the essentially unrestricted number of sera which can be tested daily, the availability of results within 24 h and the lack of interference by rheumatoid factor and by rubella-specific IgG.

Antibodies, Viral↗

Solid phase immunosorbent technique (SPIT) for the demonstration of specific IGM antibodies by hemagglutination inhibition and passive hemagglutination.

The adsorption of protein to glass and plastic has been well documented. It has also been demonstrated that the absorbed antigen or antibodies retain their activity in as much as they are able to react with a specific antibody respective specific antigen(1). Furthermore the adsorbed antignene or antibody complex may also react with a later added antigen. Anti IgM is adsorbed to the solid phase and will retain IgM from the patient's serum. If the patient's serum contains specific IgM this will react with the specific antignene. If the antigen is a hemagglutinin it will combine with the specific IgM and therefore added erythrocytes will not hemagglutinate. If the antigen is coated on erythrocytes the coated erythrocytes will combine with the specific IgM and hemagglutination will take place. The reaction is schematically demonstrated in Fig. 1.

Erythrocytes↗

New paper enzyme-linked immunosorbent technique compared with microimmunofluorescence for detection of human serum antibodies to Rickettsia tsutsugamushi.

A new paper enzyme-linked immunosorbent assay was developed for the screening and titration of human serum antibodies against the scrub typhus rickettsia, Rickettsia tsutsugamushi. The objective [corrected] was to provide a relatively simple method for antibody screening which required neither sophisticated laboratory equipment nor a high degree of technological skill. The technique develops an enzyme product from filter paper saturated with a 5-aminosalicylic acid substrate and enzymatically reacted with a commerically available anti-human immunoglobulin G peroxidase conjugate. The product of the enzymatic reaction can be interpreted visually. Comparison of 351 human sera tested by the immunofluorescent and paper enzyme-linked immunosorbent assays against a three-antigen pool of the Karp, Kato, and Gilliam strains of R. tsutsugamushi demonstrated an agreement of 96%. The sensitivity of the paper enzyme-linked immunosorbent assay as compared to immunofluorescence was 98.2%, and the specificity was 94.4%.

Antibodies, Bacterial↗

Detection and quantification of foot and mouth disease virus by enzyme labelled immunosorbent assay techniques.

Enzyme labelled immunosorbent assays (ELISA) have been developed to detect and quantify foot and mouth disease (FMD) virus using flexible plastic microtitre plates. The methods were successful for the specific detection of FMD virus and were 50 to 100 times more sensitive than the complement fixation test. The application of the ELISA techniques to FMD virus typing and subtyping, and to the assay of antigen concentration during manufacture of vaccines is discussed.

Aphthovirus↗

Comparison of immune response against Orientia tsutsugamushi, a causative agent of scrub typhus, in 4-week-old and 10-week-old scrub typhus-infected laboratory mice using enzyme-linked immunosorbent assay technique.

Using the indirect enzyme-linked immunosorbent assay (ELISA) technique, we compared the immune response of specific antibodies (IgM and IgG) in serum samples of scrub typhus-infected and non-infected mice against Orientia sutsugamushi (a causative agent of scrub typhus). Two different age groups (4-week-old and 10-week-old) of ICR laboratory mice were infected with O. tsutsugamushi by the animal passage procedure. Serum samples were detected for scrub typhus-specific antibodies using ELISA technique. Results from determining the presence of IgM and IgG in the serum samples obtained from scrub typhus-infected mice showed that the IgG was first detected on day-13 after the initial infection in both the 4-week-old and 10-week-old mice. The IgG titer levels of both groups were not significantly different. Although the presence of IgM in the in serum samples obtained from scrub typhus-infected mice was first detected on day 13 in the 4-week-old mice and on day-12 in the 10-week-old mice, the IgM titer in the 4-week-old mice was slightly lower than in the 10-week-old mice. Clinical observations of the scrub typhus-infected mice showed that the older mice become ill on day 9 whereas the younger mice exhibited the symptoms on day 12. Considering the earlier detection and slightly higher level of specific IgM antibody, it could be interpreted that the older mice may appear to have responded against O. tsutsugamushi faster than the younger mice.

Aging↗

Detection of transmissible gastroenteritis coronavirus antigens by a sandwich enzyme-linked immunosorbent assay technique.

A new sandwich enzyme-linked immunosorbent assay, using monoclonal and polyclonal antibodies, was developed to detect transmissible gastroenteritis virus antigens from cell culture and from intestinal wash or feces obtained from experimentally infected pigs. This technique was shown to be suitable for the detection of virulent field strain unadapted to cell culture. Cross reactions had not been observed with other enteric pathogens, rotavirus, porcine epizootic diarrhea virus, and Escherichia coli.

Animals↗

Diagnosis of influenza A virus infections by detection of specific immunoglobulins M, A, and G in serum.

The value of immunoglobulin M (IgM) detection in the early diagnosis of influenza A was examined in a prospective study during an outbreak of influenza A/Philippines/2/82 (H3N2) virus infection in February and March 1986. The IgM response was investigated in sera from 64 adults with influenzalike symptoms; we found a fourfold rise in titer or high titers (greater than or equal to 80) of influenza A virus antibodies by the complement fixation test. The IgM response, analyzed by the hemadsorption immunosorbent technique, was compared with the IgG and IgA responses analyzed by an indirect enzyme-linked immunosorbent assay and the hemadsorption immunosorbent technique, respectively. Antigen detection in nasopharyngeal secretions by immunofluorescence was performed for all patients on admission to hospital. Specific IgM was detected in 86% (55 of 64) of the patients with influenza A. In sera from 36% (18 of 64) of the patients it was detected already on admission. Influenza A virus antigen was detected in nasopharyngeal cells by immunofluorescence on admission in 53% (34 of 64) of the patients. A combination of immunofluorescence and IgM results gave a significantly higher diagnostic rate, 69% (P less than 0.01), on admission than did each of the two tests separately. An IgA serum antibody response was seen in 76% (48 of 64) of the cases but did not contribute to any increase in the diagnostic rate. IgM detection by the hemadsorption immunosorbent technique was found to be a valuable supplement for the diagnosis of influenza A in an early phase of the disease.

Aged↗

Comparison of different methods for the detection of rubella-specific IgM antibodies.

The rubella specific IgM titer in the serum specimens originating from healthy persons and from patients with clinical signs of rubella infection was determined by hemagglutination inhibition or hemagglutination reduction after IgM separation with the following methods: (a) density gradient centrifugation; (b) polyacrylamide agarose gel chromatography; (c) ion exchange chromatography with diethylaminoethyl cellulose columns; (d) solid-phase immunosorbent technique using microplates; (e) solid-phase immunosorbent technique using polyacrylamide microimmunobeads. Alternatively, we removed IgG and IgA by the use of protein A, anti-IgG, and anti-IgA, covalently coupled to controlled-pore glass (f). The titers obtained by the different methods showed qualitatively good correlations when combined with mercaptoethanol reduction. The quantitative measurement of specific IgM titers, however, revealed a lower sensitivity of column chromatography and methods of removal of IgG/IgA.

Antibodies, Viral↗