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Diagnosis of invasive aspergillosis by passive hemagglutination assay of antibody.

Sheep red cells treated with concanavalin A and sensitized with a partially purified aspergillus antigen were used to detect antibody to Aspergillus by passive hemagglutination (PHA). Sera from eight patients with aspergillomas or allergic aspergillosis had PHA titers of greater than or equal to 1:800 and antibody detectable by immunodiffusion (ID). Of 122 hospitalized cancer patients without invasive aspergillosis, 118 had titers of less than or equal to 1:80, 86 of < 1:10, two of 1:160, and two of 1:320. None had antibody by ID. Antibody was detectable by PHA in sera from 12 of 14 healthy microbiology laboratory workers. Of 55 cancer patients who had sera available for testing within two weeks before diagnosis of invasive aspergillosis, 18 patients seroconverted: 13 by both PHA and ID, two by PHA alone, and three by ID alone. PHA titers rose from < 1:10 to between 1:40 and 1:1,280. In immunosuppressed patients who were at risk of developing invasive aspergillosis, the appearance of antibody correlated with the diagnosis of invasive aspergillosis.

Antibodies, Fungal↗

Mannose-resistant hemagglutination and P receptor recognition of uropathogenic Escherichia coli isolated from adult patients.

Adhesions of 211 strains of uropathogenic Escherichia coli and 19 strains of normal fecal E. coli were characterized by patterns of agglutination with human erythrocytes, Saccharomyces cerevisiae, and horse erythrocytes coated with the P blood-group receptor (P). Mannose-resistant (MR) hemagglutination was significantly associated with P agglutination (P less than .001). E. coli expressing MR and/or P (MR/P) agglutinins concurrently with mannose-sensitive (MS) agglutinins predominated in all clinical categories. The highest percentage of E. coli demonstrating MR/P agglutinins, in the absence of MS agglutinins, was recovered from patients with acute pyelonephritis (35%) compared with percentages of patients with chronic pyelonephritis (13%), asymptomatic bacteriuria (16%), cystitis (11%), and normal fecal control E. coli (11%). Sixty-nine percent of E. coli isolates causing acute pyelonephritis agglutinated P-coated horse erythrocytes compared with only 11% of the fecal isolates. Strains expressing MR/P agglutinins (in the absence of MS agglutinins) isolated from patients with acute pyelonephritis, chronic pyelonephritis, and asymptomatic bacteriuria were significantly associated with the presence of antibody-coated bacteria in patients' urine sediments (P less than .010), an observation indicative of an immune response associated with bacterial invasion of host tissues.

Adhesins, Escherichia coli↗

MR/K hemagglutination of Providencia stuartii correlates with adherence to catheters and with persistence in catheter-associated bacteriuria.

Providencia stuartii was the most prevalent bacterial species isolated, for one year, from weekly urine specimens from 51 long-term catheterized patients. Significantly more strains causing bacteriuric episodes of long duration expressed MR/K (mannose-resistant/Klebsiella-like) hemagglutination (74%) than did those causing episodes of short duration (26%; P = .004). Isolates expressing MR/K hemagglutinin bound in higher numbers to catheter material (P = .023) than did those not expressing this hemagglutinin. Significantly more strains causing bacteriuric episodes of short duration expressed the mannose-sensitive (MS) hemagglutinin (43%) than did those causing episodes of long duration (7%; P = .014). Isolates expressing MS hemagglutinin bound significantly more 125I-labeled Tamm-Horsfall protein (THP) than did isolates not expressing this hemagglutinin (P = .0001). Our results indicate that MR/K hemagglutinin plays an important role in the ability of P. stuartii to persist and suggest that MR/K adheres to the catheter. Conversely, MS hemagglutinin binds to THP and may prevent persistence of P. stuartii in the catheterized urinary tract.

Aged↗

Fimbria-mediated adherence and hemagglutination of Haemophilus influenzae.

The gram-negative bacterium Haemophilus influenzae expresses morphologically and functionally distinct types of fimbriae, of which the LKP fimbriae mediate hemagglutination and adherence to human epithelial cells but hamper mucosal invasion. Therefore, the both in vivo and in vitro observed fimbrial phase variation may contribute to the pathogenesis of the infection. The existence of greater than 14 LKP serotypes hampers vaccine development based on fimbriae, since a monovalent fimbria vaccine confers protection against only the homologous strain. Cloning of the fimbrial genes in Escherichia coli results in the expression of morphologically intact fimbriae. Analysis of the cloned DNA indicates that a fimbrial gene cluster is necessary for formation of complete fimbriae and for fimbria-mediated adherence. The gene encoding the subunit is highly conserved among H. influenzae and belongs to the family of E. coli fimbriae. The phase variation is transcriptionally regulated by variation of the length of the reiterated sequence that forms the promoter region of the subunit gene.

Animals↗

Comparability and reliability of ELISA, immunofluorescence, and indirect hemagglutination assays for Trypanosoma cruzi and Trypanosoma rangeli.

Serum samples and filter paper spot samples from 48 adults were assayed in three laboratories for Trypanosoma cruzi and Trypanosoma rangeli by ELISA, immunofluorescence (IF), and indirect hemagglutination (HA) in a controlled, blind experiment. IF (serum samples) attained higher levels of comparability necessary for validity than did ELISA (serum samples) in these laboratories. ELISA was inferior to IF and HA (spot samples) in testing for T. cruzi. ELISA was not reliable (intrasubject r = .51) and was not in agreement between two laboratories (kappa = .09). The experiment indicates that ELISA is not the best choice of assay for T. cruzi in epidemiologic surveys. Assays for T. rangeli by ELISA and IF lacked comparability to the extent that those two techniques could not be considered useful for T. rangeli.

Adult↗

Prevalence of hemagglutination inhibition antibody to current strains of the H3N2 and H1N1 subtypes of influenza A virus in sera collected from the elderly in 1976.

Sera were collected in 1976 from 182 individuals born between 1876-1935, who included patients in a large local nursing home in Orange County, California, and patients and staff at the University of California, Irvine Medical Center. Sera were treated with receptor-destroying enzyme and assayed for hemagglutination inhibition (HI) antibody to recent strains of influenza A virus. The antigens tested were: A/Victoria/3/76 and A/Texas/1/77 for subtype H3N2; A/New Jersey/8/76, A/X53 and A/Swine/1976 for the A/swine influenza virus-like strains of subtype H1N1; and A/Brazil/11/78, A/Fort Monmouth/1/47 and A/Francis Warren/1/50 for the A/Fort Monmouth/47 virus-like strains of subtype H1N1. For individuals grouped by five- and 10-year intervals of the year of birth, the geometric mean and standard error for HI antibody to each antigen were calculated and plotted vs. date of birth. The results indicate that for a majority of individuals born between 1876-1900, the level of HI antibody would be protective against H3N2 but not against the strains of H1N1. In addition, a majority of individuals born between 1900-1925 had levels of antibody that should be protective against both H3N2 and the A/swine influenza virus-like strains of H1N1 but not against the A/Fort Monmouth/47 virus-like strains of H1N1. These findings could be important when deciding on the antigenic composition of vaccines to be used in those age groups that are most vulnerable to the complications of influenza. In addition, data were obtained that suggest that the A/Fort Monmouth/47 virus-like strains of H1N1 recycle in 35-40 years rather than in 65-70 years as has been demonstrated previously for H2N2 and H3N2.

Aged↗

Prevalence of antibody to serogroups 1-4 of Legionella pneumophila: a seroepidemiologic study using the indirect hemagglutination test.

An indirect hemagglutination test was used to determine the prevalence of antibody to serogroups 1-4 of Legionella pneumophila in sera from 1200 apparently healthy Michigan residents. Serogroup 1 was the most prevalent; 71 (11.8%) of 600 sera collected during the winter months (January-April, 1980) and 131 (21.8%) of 600 sera collected during the summer period (July-September, 1980) had serogroup 1 titers. This seasonal difference was independent of sex and was statistically significant in four of six age groups studied. A trend toward decreasing prevalence in the 50-59 years and 60 years or older age groups was noted in the winter sample and was statistically significant in the summer study. Prevalence of antibody to serogroups 2, 3, and 4 was significantly lower, and was associated with serogroup 1 reactivity.

Adolescent↗

A 35 kDa mannose-binding lectin with hemagglutinating and mitogenic activities from "Kidachi Aloe" (Aloe arborescens Miller var. natalensis Berger).

A novel lectin was isolated from the leaf skin of "Kidachi Aloe" (Aloe arborescens Miller var. natalensis Berger) by sequential chromatographies on Sephadex G-25 gel filtration, DEAE ion exchange, and Superdex 75 gel filtration columns. The native lectin exhibited a molecular mass of about 35 kDa on both gel filtration on a Superdex 75 column and native-PAGE under nonreducing conditions. SDS-PAGE in the presence or absence of beta-mercaptoethanol revealed two distinct peptides with molecular masses of about 5.5 and 2.3 kDa, respectively, in addition to a major 9.2 kDa subunit, indicating the presence of a partially processed subunit. The N-terminal amino acid sequence of the intact subunit showed homology with that of snowdrop lectin. The native lectin showed hemagglutinating activity toward rabbit but not human and sheep erythrocytes, and specifically bound to mannose like snowdrop lectin did, indicating that the Aloe and snowdrop lectins are structurally and functionally similar proteins. In addition, the native lectin showed strong mitogenic activity toward mouse lymphocytes.

Aloe↗

Gangliosides as paramyxovirus receptor. Structural requirement of sialo-oligosaccharides in receptors for hemagglutinating virus of Japan (Sendai virus) and Newcastle disease virus.

A sensitive assay system for receptor activity of gangliosides to paramyxovirus was developed. This system involves incorporation of gangliosides into neuraminidase-treated chicken erythrocytes (asialoerythrocytes) followed by estimation of virus-mediated agglutination and hemolysis. The asialoerythrocytes coated with I-active ganglioside (Sia alpha 2-3Gal beta 1-4GlcNAc beta 1-3(Gal alpha 1-3Gal beta 1-4GlcNAc beta 1-6)Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc beta 1-Cer) were effectively agglutinated by hemagglutinating virus of Japan (HVJ, Sendai virus). The hemolysis of the asialoerythrocytes mediated by HVJ was restored to the highest level by labeling the cells with gangliosides possessing lacto-series oligosaccharide chains, i.e., I-active ganglioside, N-acetylneuraminosylparagloboside (SiaPG(NeuAc)), and i-active ganglioside (Sia alpha 2-3Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc beta 1-Cer). The specific receptor activity of ganglioside GD1a possessing a gangliotetraose chain was lower than those of the gangliosides described above. Gangliosides GM3, GD3, GM1a, GD1b, SiaPG(NeuGc) showed little effect on the restoration of HVJ-mediated hemolysis. On infection with Newcastle disease virus (NDV), the highest specific restoration of lysis was found in chicken asialoerythrocytes coated with SiaPG(NeuAc or NeuGc) and GM3(NeuAc or NeuGc), whereas those coated with I-active ganglioside, GD3, GM1a, and GD1b showed very low NDV-mediated hemolysis. The above results indicate that the determinants of receptor for HVJ contain sialylated branched and/or linear lacto-series oligosaccharides carried by I,i-active gangliosides and SiaPG(NeuAc) and sialosylgangliotetraose chain carried by GD1a. The determinants for NDV are carried by SiaPG(NeuAc or NeuGc) containing linear lacto-series oligosaccharide and GM3(NeuAc or NeuGc). The absence of detectable binding of free oligosaccharides obtained from I-active ganglioside and sialoglycoprotein GP-2 isolated from bovine erythrocyte membranes as HVJ receptor (Suzuki, Y., et al. J. Biochem. (1983) 93, 1621-1633; (1984) 95, 1193-1200) indicates that HVJ recognizes the sialooligosaccharides oriented out of the lipid bilayer in the cell membranes where the hydrophobic ceramide or peptide backbone of the receptor is integrated.

Animals↗

Evaluation of the indirect hemagglutination technique for study of Trichomonas vaginalis infections, particularly in men.

The indirect hemagglutination (IHA) technique was evaluated for use in the serologic study of infection with Trichomonas vaginalis. The IHA test showed that sera from 88% of women attending a venereal disease clinic had antibody to T. vaginalis. The antibody frequency and titers were highest in women who had documented infections due to T. vaginalis. Serologic and cultural evidence of recent or active trichomonal infection was found in 11% of 85 men who had nongonococcal urethritis, but was absent in a control group of 27 men. Clinical findings in ten men with symptomatic genitourinary trichomoniasis are described; all but one had relatively high (greater than or equal to 1:80) IHA antibody titers. Antibody to T. vaginalis was found significantly (P less than 0.005) more often in sera from women than in sera from men in an apparently healthy group of individuals between the ages of 1 and 20 years. The IHA test appears potentially useful for the diagnosis of trichomoniasis in men and in the seroepidemiology of infections due to T. vaginalis.

Evaluation Studies as Topic↗

Seven-year follow-up study of rubella syndrome in Ryukyu with special reference to persistence of rubella hemagglutination inhibition antibodies.

Rubella hemagglutination inhibition (HI) antibodies in 266 children with rubella syndrome born in 1965 in the Ryukyu Islands and their mothers were followed for seven years. Titers of rubella HI antibody in the mothers declined slowly, while those in the children declined rapidly up to 40 months of age. Thereafter decline of titers became extremely slow and only seven cases (three per cent) became seronegative for rubella HI antibody. Rubella HI antibody titers seemed to have no particular correlation to the severity of clinical manifestations.

Adult↗

Improved reversed passive hemagglutination for simple and rapid detection of staphycococcal enterotoxins A approximately E in food.

Detection and identification of staphylococcal enterotoxins in food or culture filtrates were performed using the reversed passive hemagglutination (RPHA) technique, with formalized sheep red blood cells (FSRBC) sensitized with immunologlobulins of anti-A, B, C, D, and E rabbit hyperimmune sera fractionated by affinity chromatography. The FSRBC sensitized with anti-A approximately E immunoglobulins showed a high level of reactivity and specificity in RPHA, against homologous types of purified enterotoxins and culture filtrates of toxin-producing strains. No non-specific reactions with various ingredients in foods nor cross-reactions among enterotoxin types were observed. The minimum amount of enterotoxins in foods detected by RPHA was calculated to be 0.01 micorgram/g without concentration, and the recovery rate of experimentally added toxins was calculated to be about 80%. Under routine laboratory practice, detection and identification of enterotoxins from incriminated foods of five food poisoning outbreaks were performed by RPHA within 3 hr after reception of the specimens. Among them, three were determined to be enterotoxin A food poisoning, one to be toxin C and the rest to be intoxication of A and D. The concentrations of the toxins was between 0.014 and 3.65 microgram per gram of food.

Disease Outbreaks↗

Hemagglutination by pilus antigen 987P of enterotoxigenic Escherichia coli.

The hemagglutination (HA) by pilus antigen 987P of an enterotoxigenic Escherichia coli strain 987 was examined using fresh and glutaraldehyde (GA)-fixed erythrocytes (RBC) of various animals. Only when GA-fixed RBC was employed, a strain 987 exhibited striking HA activities. This was also demonstrated by using latex heads sensitized with the 987P antigen. The 987P-specific antiserum inhibited HA of strain 987 and 987P sensitized latex beads against GA-fixed RBC. We concluded that HA of strain 987 against GA-fixed RBC was specifically associated with the presence of 987P pilus antigen but do not exclude a possibility that adhesin is distinct from pili antigen.

Adhesins, Escherichia coli↗

Hemagglutination activity and localization of Fc receptor of group A and G streptococci.

The IgG-Fc binding activity and binding sites on the cell surface of streptococci, strains AR1 (group A) and G148 (group G), and Staphylococcus aureus strain Cowan I were examined by hemagglutination (HA) and immunoelectron microscopic methods. No distinct difference was observed in the HA activity among these three strains. However, the strains differed in the distribution of Fc receptor. Cowan I cells (having protein A) were heavily covered with two layers of ferritin particles, whereas AR1 cells were heavily covered with a single, rough layer of ferritin particles. G148 cells (having protein G) were labeled with a relatively thin, rough ferritin layer. The trypsin susceptibility of the Fc receptors of the AR1 strain was much higher than that of the G148 strain. These results suggest that both streptococcal strains are distinctly different in the arrangement or in the conformation of the Fc receptor from the Cowan I strain. It is also suggested that the Fc receptor molecules of the streptococcal strains differ from each other not only in conformation but also in trypsin susceptibility.

Hemagglutination↗

Cell-surface properties of enterotoxigenic and cytotoxic Salmonella enteritidis and Salmonella typhimurium: studies on hemagglutination, cell-surface hydrophobicity, attachment to human intestinal cells and fibronectin-binding.

Thirteen Salmonella enteritidis and S. typhimurium strains with smooth or rough colony morphology were investigated for their surface properties based on hemagglutination (HA), hydrophobicity, and fibronectin-binding profiles. The strains showed 5 different patterns of HA which was mannose-sensitive. The rough strains possessed comparatively greater number of fimbriae than the corresponding smooth strains and also attached to human intestinal cells in greater numbers. The Salmonella strains used in this study interacted with fibronectin and its 29-kDa N-terminal fragment to varied extents. These properties may be helpful in broadening the prospective interaction capabilities of Salmonella organisms with the host surfaces.

Bacterial Adhesion↗

Single-serum diagnosis of recent rubella infection with the use of hemagglutination inhibition test and enzyme-linked immunosorbent assays.

Single-serum diagnosis of recent rubella infection was attempted with the use of hemagglutination inhibition (HI) test and two commercially available enzyme-linked immunosorbent assays (ELISAs). The period during which IgM antibody was detectable by IgM capture ELISA was 4 to 30 days after the onset of rash. Rubella IgG ELISA values relative to HI titer were lower in the sera from the patients with recent infection than in the sera from the subjects with remote infection. IgM ELISA and the combined use of IgG ELISA and HI test are two useful methods of single-serum diagnosis of recent rubella infection.

Antibodies, Viral↗

Passive hemagglutination assays for the detection of antibodies to herpes viruses.

A simple and effective method for the detection of antibodies to herpes simplex virus (HSV), human cytomegalovirus (HCMV) and varicella-zoster virus (VZV), has been established using the passive hemagglutination assay (PHA) in combination with viral specific glycoproteins. The results obtained with the PHA were compared with those from neutralization (NT) and complement fixation (CF) tests. The PHA test for each of the herpes viruses appears to compare favorably with the other assays tested. The specificity and sensitivity of HSV PHA to NT were 100%, whereas the specificity and sensitivity of HSV CF test to NT were 98% and 100%, respectively. For HCMV, the specificity and sensitivity of PHA to NT and PHA to CF were 100%. Similarly, the specificity and sensitivity of VZV PHA to NT were 100%. Because of the low sensitivity of the VZV CF, the sensitivity of CF to NT was 83%. Furthermore, the range of antibody titers and their absolute levels obtained in the PHAs were significantly greater than those in the NT and CF tests.

Adult↗

Production of antigenically related exocellular elastolytic proteases mediating hemagglutination by vibrios.

Exocellular proteases produced by Vibrio fluvialis, V. furnissii, V. metschnikovii and V. campbellii were characterized and compared to those of V. mimicus protease (VMP) and V. vulnificus protease (VVP). These proteases possessed both elastolytic and hemagglutinating abilities and were identified, except that of V. metschnikovii, as metalloprotease. Conversely, V. metschnikovii protease failed to exhibit some of the salient features for metalloproteases suggesting the existence of protease(s) other than metalloprotease. However, antibodies against VVP cross-reacted to these proteases and to VMP indicating antigenic relatedness amongst vibrio proteases. This study, thus, demonstrated the prevalent distributions of antigenically related proteases both in pathogenic and non-pathogenic vibrios, bringing their status as a virulence determinant into question.

Antigens, Bacterial↗