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PubMed1983

Increased frequency of ColV plasmids and mannose-resistant hemagglutinating activity in an Escherichia coli K1 population.

The expression of traits linked to pathogenicity was studied in a population of Escherichia coli K1 strains. It was found that E. coli K1 strains isolated from extraintestinal infection harbor the ColV plasmid and express mannose-resistant hemagglutinating activity type VI with a high frequency. The presence of these properties may play a role in the ability of some E. coli K1 serogroups to invade.

Bacteriocin Plasmids↗
PubMed1984

Evaluation of the hemagglutination treponemal test for syphilis.

The sensitivity and specificity of the hemagglutination treponemal test for syphilis (HATTS) was compared with the fluorescent treponemal antibody-absorption test (FTA-ABS) with 491 sera. Medical histories were obtained for 153 patients with seroreactivity in either treponemal test. Overall correlation with patient history was 96.7% for the FTA-ABS and 93.9% for the HATTS. False-negative HATTS occurred in primary, late-latent (greater than 2 years), and treated syphilis. The reproducibility of the HATTS and FTA-ABS is equivalent. HATTS is an acceptable treponemal confirmatory test for syphilis, but the FTA-ABS should be used to resolve diagnostic discrepancies.

Antibodies, Bacterial↗
PubMed1975

Comparison of the reversed passive hemagglutination with radioimmunoassay methods for hepatitis B antigen.

Radioimmunoassay for the detection of hepatitis B antigen has been accepted in many diagnostic laboratories now. The question of nonspecific positives has always been a matter of controversy. Two improved radioimmunoassay tests, namely Ausria II-125 by Abbott Laboratories and a radioimmunoassay method by Connaught Laboratories Limited (Hebria), were compared with the original Ausria 125I. Included in the comparison was the reversed passive hemagglutination test (Auscell, Abbott). Five hundred sera of clinical patients were tested. Fifty-five or 11% were found to have hepatitis B antigen. Three tests, Ausria 125I, Ausria II-125, and Hebria showed the same number of positive sera, whereas Auscell missed one. However, Ausria 125I indicated two additional false positives. Dilution experiments, however, indicated that Ausria II-125 and Hebria were the most sensitive tests, with the reversed passive hemaglutination showing the least sensitivity. Therefore, the new Ausria II-125 and the Hebria radioimmunoassay tests are preferable in view of their sensitivity and specificity.

Blood↗
PubMed1975

Indirect hemagglutination test for human antibody to typhus and spotted fever group rickettsiae.

An indirect hemagglutination (IHA) test is described that uses glutaraldehyde-stabilized erythrocytes treated with a rickettsial erythrocyte-sensitizing substance obtained from Rickettsia typhi or Rickettsia rickettsii. The serological reagent was stable for at least 3 months at room temperature and 6 months at 4 C. It exhibited group specificity and no group cross-reactivity. At a minimum dilution of 1:40, acute and early convalescent epidemic and murine typhus antisera showed 86% positive reactors, whereas similar spotted fever antisera had 74% positive reactors. In comparison with the indirect fluorescent antibody test, the IHA procedure gave lower titers but showed comparable detection of seroconversion with most paired sera. The IHA test demonstrated significantly higher titers than the complement fixation test and was more sensitive than either the complement fixation or Weil-Felix test in identifying seroconversion. No agglutination was observed when sensitized erythrocytes were tested with rodent sera known to contain rickettsial antibodies.

Agglutination Tests↗
PubMed1975

Simple procedure for the removal of nonspecific inhibitors of rubella virus hemagglutination.

The adsorption of serum lipoproteins onto an insoluble matrix of colloidal silicic acid results in the removal of nonspecific inhibitors of rubella virus hemagglutinin. The procedure can be performed in 15 min at room temperature. Comparative studies using both the dextran sulfate-CaCl2 and heparin-MnCl2 methods for removal of inhibitors demonstrated that the colloidal silicic acid procedure yielded identical hemagglutination inhibition titers. In addition, it is technically feasible to read titers below 1:8.

Adsorption↗
PubMed1984

Infectivity titration of hemorrhagic fever with renal syndrome virus: use of immune adherence hemagglutination for detection of virus growth.

Serial dilutions of hemorrhagic fever with renal syndrome viruses were inoculated into Vero-E6 cells in microplates. After 2 weeks of incubation, infected cells were disrupted by freezing and thawing, and virus antigens were detected by immune adherence hemagglutination. The infectivity titers of the virus as determined by this method were in close agreement with those obtained by the immunofluorescent antigen endpoint method. Then, a neutralization method was established. Japanese hemorrhagic fever with renal syndrome isolates, strains SR-11 and TR-352, were found to be distinct from Hantaan virus, strain 76-118, by the neutralization test.

Animals↗
PubMed1985

Comparison of measles antihemolysin test, enzyme-linked immunosorbent assay, and hemagglutination inhibition test with neutralization test for determination of immune status.

Sera were collected from 238 high-school students in Prince Edward Island for the determination of immune status before an anticipated measles outbreak. In addition, history of vaccination status and measles infection was obtained. In the subsequent outbreak, 28 students did contract measles. Specificity for hemagglutination inhibition (HI), antihemolysin (AH), and enzyme-linked immunosorbent assay (ELISA) was 100%, compared with the neutralization test. Corresponding sensitivity values for the tests were 66.0% (HI), 99.5% (AH), and 99.0% (ELISA). Predictive values for susceptibility were 26.9% (HI), 77.8% (AH), 75.7% (ELISA), 80% (neutralization), and 41.4% as determined by history of infection or vaccination. The predictive value for immunity as determined by history of previous infection or vaccination was 91.8%, compared with 100% for the four serological tests. No false-positive results were seen with any of these tests. Compared with the neutralization test, the HI test had 69 false-negative results, the AH had 1, and the ELISA test had 2. The AH and ELISA tests provided sensitive and specific alternatives to the commonly used HI test for immune status determination.

Adolescent↗
PubMed1985

Cytomegalovirus antigenic heterogeneity can cause false-negative results in indirect hemagglutination and complement fixation antibody assays.

Cord sera and antepartum maternal sera from three congenitally cytomegalovirus (CMV)-infected infants and their mothers were CMV seronegative (titer, less than 8) in a complement fixation (CF) assay with a glycine-extracted CMV AD169 antigen; sera from two of the infants and mothers were also seronegative in a commercial indirect hemagglutination (IHA) assay with AD169 antigen. In tests with their own CMV isolates propagated and made into glycine-extracted CF antigen, all were seropositive. When 108 random cord sera were assayed for CF antibody with AD169, Davis, and A antigens (A is a locally derived antigen from one of the above infants), 44 were seropositive and 54 were seronegative for all three antigens. Of the remaining 10 sera, 4 were positive for A only, 3 were positive for A and Davis only, 2 were positive for Davis and AD169 only, and 1 was positive for AD169 only. All 10 were positive when a mixture of all three antigens was used. The IHA assay with AD169 antigen was positive with only 4 of these 10 sera. These results suggest that up to 6% of sera may be misclassified as seronegative in the CF and IHA assays if only a single antigen is used.

Antibodies, Viral↗
PubMed1986

Separation of hemagglutination-inhibiting immunoglobulin M antibody to rubella virus in human serum by high-performance liquid chromatography.

High-performance liquid chromatography was successfully used to separate hemagglutination-inhibiting immunoglobulin M (IgM) rubella virus antibody from IgG rubella virus antibody in human serum. The fractionation by high-performance liquid chromatography was as effective as sucrose density gradient centrifugation in separating IgM antibody from IgG antibody.

Antibodies, Viral↗
PubMed1986

Comparison of immunofluorescence and hemagglutination inhibition tests and enzyme-linked immunosorbent assay for detection of serum antibody in rats infected with hemorrhagic fever with renal syndrome virus.

Serum antibodies against the B-1 strain of hemorrhagic fever with renal syndrome (HFRS) virus in wild and experimental rats were investigated by the indirect immunofluorescent antibody (IF) test, the hemagglutination inhibition (HI) test, and an enzyme-linked immunosorbent assay (ELISA). In the newly developed ELISA, monoclonal antibodies to the B-1 strain were applied as coating antibody. In sera from wild rats, the results obtained by the ELISA agreed quite well with those obtained by the HI test, but some serum samples that gave a negative reaction in the HI test gave a positive reaction in the IF test, although their IF titers were very low. In serum samples from experimental rats that had been kept in an animal house infected with HFRS virus, a group with high titers and a group with low or negative titers were clearly differentiated by all the tests.

Animals↗
PubMed1987

Passive hemagglutination test for enteric fever.

A passive hemagglutination (PHA) test for serodiagnosis of enteric fever was developed by sensitizing glutaraldehyde-preserved erythrocytes with lipopolysaccharide from Salmonella serogroups A, B, C, and D singly or simultaneously. The lipopolysaccharide-sensitized erythrocytes were tested with sera from 200 blood donors, 100 patients whose hemoculture was positive for Salmonella species, and 10 patients septicemic for other members of the family Enterobacteriaceae. The PHA test was positive in 90% of 28 acute-phase serum samples from patients with enteric fever from one hospital and in 93% of 72 acute-phase serum samples from another hospital. It was also positive in 100 and 60% of early- and late-convalescent-phase sera, respectively. The PHA test was negative in all patients septicemic for other members of the Enterobacteriaceae. Absorption of sera from patients with enteric fever with lipopolysaccharide from other members of the Enterobacteriaceae did not reduce PHA titers, indicating the specificity of the PHA test. Simultaneous sensitization with lipopolysaccharide from Salmonella serogroups A, B, C, and D was useful as a screening test in a limited trial with 28 acute-phase sera, 10 early-convalescent-phase sera, and 17 late-convalescent-phase sera. The PHA test is indeed a simple, sensitive, specific, and rapid test supplementing hemoculture in laboratory diagnosis of enteric fever.

Enterobacteriaceae↗
PubMed1988

Evaluation of enzyme-linked immunosorbent assay and reversed passive hemagglutination for detection of Crimean-Congo hemorrhagic fever virus antigen.

Enzyme-linked immunosorbent assay (ELISA) and a reversed passive hemagglutination (RPHA) test were evaluated for rapid detection of Crimean-Congo hemorrhagic fever (CCHF) virus antigens. Both RPHA and ELISA detected CCHF antigen in the brains of infant mice 2 to 3 days after infection, several days before the animals sickened and died. Antigen was also detected after 1 to 2 days in infected cell culture extracts and after 2 to 4 days in culture supernatant fluids. Both tests detected CCHF antigen at threshold values of approximately 2.5 log10 tissue culture infective doses per ml and were more sensitive than complement fixation, immunodiffusion, or immunofluorescence. In a comparative study on specimens from CCHF patients, virus was isolated from 38 of 49 sera and 23 of 28 patients. Antigen was detected in 20 of 49 sera (15 of 28 patients) by RPHA and in 29 of 49 sera (18 of 28 patients) by ELISA. Antigenemia was detected more frequently in fatal cases (9 of 11) than in nonfatal cases (9 of 17). Although the antigen detection assays offered a more rapid approach than infectivity assays for diagnosing CCHF, the latter test was more sensitive. The results suggest that RPHA and ELISA may be of use in rapid diagnosis of CCHF infection, particularly in severe cases, in which the danger of nosocomial spread is greatest.

Animals↗
PubMed1989

Passive hemagglutination test for detection of antibodies to human immunodeficiency virus type 1 and comparison of the test with enzyme-linked immunosorbent assay and Western blot (immunoblot) analysis.

A passive hemagglutination test (PHA) was developed for detecting antibodies to human immunodeficiency virus type 1 (HIV-1) utilizing sheep erythrocytes cross-linked with purified envelope glycoprotein (gp160) of HIV-1. In an analysis of 216 human serum samples, 100% correlation was observed in 86 reactive and 124 nonreactive serum samples between PHA and commercial enzyme-linked immunosorbent assays and Western blot (immunoblot) analysis. Serum samples from gp160-immunized chimpanzees also reacted equally well in PHA. The test is simple, rapid, and inexpensive, thus providing an alternate, quick method of detecting HIV antibodies. These advantages and the thermal stability of the reagents that are used make this an attractive alternative for detecting prior exposure of individuals to HIV-1.

Animals↗
PubMed1989

Detection of antibodies to human immunodeficiency virus type 1 in whole blood and saliva by using a passive hemagglutination test.

A passive hemagglutination test (PHA) for detecting human immunodeficiency virus type 1 antibodies in serum samples by using envelope glycoprotein (gp160)-coupled sheep erythrocytes was described earlier (M.B. Vasudevachari, K. U. Uffelman, T.C. Mast, R.L. Dewar, V. Natarajan, H.C. Lane, and N.P. Salzman, J. Clin. Microbiol. 27:179-181, 1989). In the study reported here, the applicability of the PHA test to the detection of antibodies in whole-blood and saliva samples has been investigated. We observed a 100% correlation between PHA and commercial enzyme-linked immunosorbent assay in 101 whole-blood samples and 98% correlation between PHA and reactivity to envelope proteins in Western blots (immunoblots) of 53 saliva samples. Furthermore, salivary antibodies could be detected in 19 of the 22 seropositive individuals. As in serum, antibodies to envelope proteins were widely prevalent in all the Western blot-reactive saliva samples.

Blotting, Western↗
PubMed1989

Mannose-resistant hemagglutination, enzyme-linked immunosorbent assay, and immune electron microscopy for detection of K99 fimbrial antigen in Escherichia coli from calves.

Mannose-resistant hemagglutination (MRHA) was evaluated for identification of Escherichia coli with K99 fimbriae. The sensitivity and specificity of MRHA, relative to the enzyme-linked immunosorbent assay, were 21 and 79%, respectively. Disagreement between the tests may have been due in part to separation of pili from the cells, with resultant enzyme-linked immunosorbent assay-positive, MRHA-negative tests. MRHA was not useful as the sole means for the identification of E. coli with K99 fimbriae.

Animals↗
PubMed1991

Distribution of hantavirus serotypes Hantaan and Seoul causing hemorrhagic fever with renal syndrome and identification by hemagglutination inhibition assay.

An epidemiologic evaluation of patients with hemorrhagic fever with renal syndrome from different locations in the People's Republic of China was conducted to define the prevalence of two Hantavirus serotypes, Seoul (SEO) and Hantaan (HTN). Serum specimens were collected between 5 and 14 days after the onset of illness and were tested for antibodies by both hemagglutination inhibition (HI) and plaque reduction neutralization (PRN). By the HI test, the geometric mean titer (GMT) of antibodies to SEO in the sera from individuals from Kaifeng City of Henan Province was five times higher than that to HTN. In contrast, by the HI test, the sera from individuals from Jiande County of Zhejiang Province had a GMT of antibodies to HTN that was seven times higher than that to SEO. In the sera from individuals from Shanghai, only a twofold difference was observed in HI antibody titers to the two hemagglutinins by the HI test, with that to HTN being higher than that to SEO. By the PRN test, the GMT ratios of antibody between HTN and SEO strains from individuals in Kaifeng, Jiande, and Shanghai were found to be 1:13, 14:1, and 2:1 respectively. A close correlation (r = 0.8219) and concordance rate (78.3%) were observed between the PRN and HI tests for the identification of the serotypes of individual cases of hemorrhagic fever with renal syndrome. The hantavirus serotypes from individuals in Kaifeng and Jiande were identified as predominantly SEO and HTN, respectively, and those from individuals in Shanghai had an indeterminant serotype defined by these two techniques. The HI test appears to be a simple and reliable way of determining the predominant hantavirus that causes HFRS in a given geographic area.

Antibodies, Viral↗
PubMed1976

Evaluation of a rubella hemagglutination inhibition test system.

Two systems for measurement of rubella hemagglutination inhibition antibodies were compared. One had chick erythrocytes as indicator and the other, which is available in kit form ("Rubindex," Ortho-Diagnostics), uses human group O erythrocytes. Correlation to within one dilution of each other was noted in 92% of the tests. The Rubindex system has the advantage of longer shelf life of reagents and also does not require one of the absorption steps necessary for the other method, thus saving technologist's time.

ABO Blood-Group System↗