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Desorption of porcine parvovirus from aluminum hydroxide adjuvant with subsequent viral immunoassay or hemagglutination assay.

Cell culture fluids containing porcine parvovirus were adjuvanted with varying concentrations of aluminum hydroxide gel. Adsorption of virus and total protein to adjuvant was proportional to adjuvant concentration. Desorption of virus and protein from the adjuvant in substantial, reproducible quantities was achieved by washing adjuvanted preparations with 1.2 M potassium phosphate, followed by dialysis and concentration of wash fluids. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of unadjuvanted viral fluids, supernatants of adjuvanted mixtures, and desorptive washings of the corresponding adjuvant pellets revealed no qualitative differences in protein banding patterns. Desorbed virus was quantitated by a hemagglutination assay, or by an enzyme-linked immunoassay employing a monoclonal anti-porcine parvovirus antibody. Virus desorption and subsequent assays permitted in vitro estimation of virus content in adjuvanted porcine parvovirus preparations. This approach may be useful in estimating the antigen content of inactivated aluminum hydroxide adjuvanted veterinary vaccines and reducing the extent of required in vivo testing.

Adjuvants, Immunologic↗

Stratum corneum antibodies detected by hemagglutination are not directed against keratin intermediate filaments.

Autoantibodies to stratum corneum (SC) occur in virtually all normal adult human sera. These antibodies may be directed against various antigens of the SC. They have been detected by indirect immunofluorescence, passive hemagglutination (HA), immune adherence, and most recently by enzyme immunoassay and immunoblot methods. The purpose of our study was to examine whether antibodies to SC antigens as detected by passive HA are similar to the keratin intermediate filament (KIF) reactive antibodies. SC antigen preparation was prepared from psoriatic scales by the trypsin-phenol-water (TPW) extraction method. KIFs were prepared by 8 M urea extraction of normal callus or psoriatic scales. The anti-SC antibody titers of normal human sera were determined by passive HA before and after absorption with TPW-SC antigen preparation and upon absorption with KIFs. Similarly, titers of anti-KIF antibodies were determined on absorbed and unabsorbed sera by immunoblot assay. The results of this study indicate that the absorption of the sera with KIFs did not affect the titer of antibodies to TPW-extractable SC antigens whereas the titer of KIF antibodies dropped. KIF-reactive antibodies, on the other hand, were not affected by absorption with TPW-SC antigen, whereas the latter absorbed out the corresponding reactive antibodies. These results indicate that antibodies directed against SC antigen are different from the KIF-reactive antibodies.

Amino Acids↗

An attempt of immunodiagnosis by means of mixed passive hemagglutination (MPHA) test for human pulmonary dirofilariasis.

The mixed passive hemagglutination (MPHA) test was used for the first time in an attempt to diagnose human pulmonary dirofilariasis in a comparison with the results of ELISA. Eight of nine patients tested with the MPHA test were positive and one was false-positive; these results coincided exactly with those of ELISA. Qualitative agreement between the MPHA test and ELISA was 90.4% of 374 human sera. The present study indicates that the MPHA test seems to be a promising diagnostic tool for human pulmonary dirofilariasis.

Adult↗

Efficacy of the urinary hemagglutination test (Higonavis) and enzyme immunoassay (Ovustick) in detection of the spontaneous luteinizing hormone surge in an in vitro fertilization/gamete intrafallopian transfer (IVF/GIFT) program.

Fifty-eight treatment cycles in an in vitro fertilization/gamete intrafallopian transfer (IVF/GIFT) program were studied to compare the efficacy of two urinary methods, hemagglutination test (Higonavis) and enzyme immunoassay (Ovustick), in detection of spontaneous luteinizing hormone (LH) surge. If an isolated rise in urinary LH level was taken as indicative of LH surge, the false-positive rate was 36.7% for Higonavis and 10.2% for Ovustick. The difference was statistically significant (P less than 0.001). If only a sustained rise in urinary LH was taken to indicate LH surge, the false-positive rate was 6.1% for Higonavis and 0% for Ovustick. In the seven cycles with a spontaneous plasma LH surge, there was a positive correlation between the plasma LH levels and the two urinary assay methods in six cycles (85.7%). Compared to plasma LH, there was a mean delay of 17.4 hr by the Higonavis test and 15.6 hr by the Ovustick test. If a sustained rise in urinary LH levels was taken as indicative of LH surge, both methods are quite accurate but the Ovustick appeared to be more specific.

Evaluation Studies as Topic↗

Antigenic relationships between avian paramyxoviruses. I. Quantitative characteristics based on hemagglutination and neuraminidase inhibition tests.

Comprehensive hemagglutination inhibition (HI) and neuraminidase inhibition (NI) cross reaction tests were performed using 8 of 9 serotypes of avian paramyxoviruses (PMV). The studies were designed as full scale repeating experiments which permitted an adequate statistical treatment and elaboration of quantitative criteria of antigenic kinship. The results have shown diverse antigenic relationships between different avian paramyxovirus (PMV) serotypes which were asymmetric in some cases. The antigenic relationships found by HI test did not always parallel those found by NI tests. The antigenic inter-relationships have been displayed quantitatively in a diagram. This has given a basis for some suggestions concerning: the independent antigenic drift of the HA and Nase antigenic sites of hemagglutinin-neuraminidase (HN) glycoprotein of avian PMVs; a tentative subdivision of the whole group of avian PMVs into two subgroups: the first including PMV-2 and PMV-6 serotypes and the second including PMV-1, PMV-3, PMV-4, PMV-7, PMV-8 and PMV-9 serotypes; the conception that genomic material coding for the HN glycoprotein consists of a "common-to-all-the-PMVs" portion and a "serotype-specific" portion, on one hand, and of a "conserved" portion and a "variable" portion, on the other; the ratios between the portions have been shown to be different for, at least, certain PMV serotypes; the evolutionary pathways of the avain PMV HN antigenic drift.

Animals↗

Feline herpesvirus type 1 glycoproteins eliciting virus neutralizing and hemagglutination-inhibiting antibodies.

Monoclonal antibodies (MoAbs) to feline herpesvirus type 1 (FHV-1) were produced to identify the virus-specified immunogenic proteins. When antigens recognized by the MoAbs were investigated by immunoblot and immunoprecipitation assays using FHV-1-infected cell lysate, four groups of immunogenic proteins were identified. MoAbs directed against 60 kDa, 113 kDa, and 143 kDa/108 kDa glycoproteins had virus neutralizing activities, but those against 170 kDa protein did not. Furthermore, MoAbs to the 60 kDa glycoprotein also showed hemagglutination-inhibition activity, indicating that this glycoprotein might be the hemagglutinin.

Animals↗

Identification of lipid components of human serum lipoproteins involved in the inhibition of Sindbis virus infectivity, hemagglutination, and hemolysis.

Human serum high density lipoproteins (HDL), low density lipoproteins (LDL) and very low density lipoproteins (VLDL) were isolated and tested for their ability to inhibit Sindbis virus infectivity, hemagglutination and hemolysis. VLDL and LDL produced a strong reduction on both viral infectivity on Vero cell monolayers and attachment and fusion with erythrocytes, whereas HDL appeared to be only a weak inhibitor. Lipid and protein components were extracted from each class of lipoproteins to identify the molecules responsible for the inhibiting activity. Only the lipid moiety was found to inhibit Sindbis virus biological activities. Among the individual lipid components of lipoproteins, neutral lipids (cholesterol, oleic acid and palmitic acid) and negatively charged phospholipids (phosphatidylserine and phosphatidylinositol) and glycolipids (GM 3 ganglioside and cerebroside sulphate) were able to neutralize the virus suggesting that either hydrophobic or electrostatic interactions are involved in the inhibition.

Apoproteins↗

Comparison of indirect hemagglutination and 51chromium release tests for detection of Herpes Simplex Virus type 1 and 2 antibodies in patients with recurrent Herpes infections.

Indirect hemagglutination and 51Cr release tests (IHAT and 51-CRT respectively) were compared in patients with recurrent herpes simplex virus (HSV) infections from whom HSV-1 or HSV-2 was isolated. Both tests were equally sensitive and specific in detecting HSV antibodies. However, IHAT was more specific in detecting homologous HSV antibody response in patients with recurrent HSV-2 infections. Past infections with HSV-1 in the patients with dual infections were detected by determining HSV-type specific antibodies by inhibition of IHAT. Cross absorption studies showed that the antibody reactivity measured by the two tests was qualitatively and quantitatively different. Nevertheless, IHAT has been found to be more appropriate test for seroepidemiologic studies of HSV-2 infections because of its specificity, rapidity and less cost, whereas, 51-CRT appears to measure antibodies against recent and more predominant type of infecting HSV.

Antibodies, Viral↗

Purification and hemagglutinating properties of egg drop syndrome 1976 virus.

We purified three populations of virus particles, F7, F9 and F17, with buoyant densities of 1.34, 1.33 and 1.29 g/ml, respectively, in CsCl equilibrium density gradients from cultures of chick embryo liver cells infected with the H-162 strain of the virus of egg drop syndrome 1976. F9 particles were infectious complete virions and most F17 particles were empty particles. F7 particles were less infectious, and had little capacity of hemagglutination (HA). HA titers were the same at 4 degrees and 37 degrees C and maximal between pH 6.4 and 8.4 and ionic strength from 0.14 to 0.54 M of NaCl. HA titer was inversely proportional to erythrocyte concentration. Potassium periodate destroyed markedly the infectivity of the virus and partially its HA activity at 37 degrees C. HA activity was stable at 56 degrees C or lower temperatures and destroyed at 80 degrees C. Trypsin, alpha-chymotrypsin, papain, ficin and neuraminidase had no effect on HA activity. Alpha-chymotrypsin destroyed the receptor for the virus on chicken erythrocytes, whereas trypsin and neuraminidase did not affect the receptor.

Adenoviridae↗

Changes of the rotavirus concentration in faeces during the course of acute gastroenteritis as determined by the immune adherence hemagglutination test.

Concentrations of rotavirus were studied in serially sampled faecal specimens from 55 patients with acute gastroenteritis, utilizing the immune adherence hemagglutination (IAHA) test. Rotavirus antigen was detected in 45 (82%) out of 55 cases. In the faeces which were positive for rotavirus antigen, the concentrations reached their maximum titer shortly after the onset of the illness and diminished gradually until the 9th day.

Acute Disease↗

Hemagglutination of neuraminidase-treated human erythrocytes by Leishmania enriettii infected guinea-pig sera.

Sera from guinea-pigs infected with the protozoan parasite Leishmania enriettii showed higher hemagglutination (HA) titres for neuraminidase treated human erythrocytes than those of normal guinea-pig sera. This HA activity was associated mostly with the 19S fraction of the immune serum and could be absorbed out with an antigenic fraction of the parasite membrane. Antigenic determinants involved in this HA reaction consisted of, at least, beta-D-galactosyl or lactosyl residues.

Animals↗

Adherence and hemagglutination of Corynebacterium group D2.

Sixty-one strains of Corynebacterium group D2 were examined for their ability to adhere to human uroepithelial cells and to agglutinate human and guinea-pig erythrocytes. Strains were isolated from samples of two origins: urine of bacteriuric patients and healthy skin of patients without urinary infection. In addition, the isolates were examined by scanning and transmission electron microscopy. Heavy adherence to the uroepithelial cells but weak hemagglutination were noted. No statistical association was demonstrated between the adherence and the origin of the strains (65.2% of urinary isolates and 80% of healthy skin isolates were adherent). On transmission electron microscopy, a close association was observed between adherent bacteria and cells on thin sections and only few strains were piliated with negative staining. These results do not support a role of adherence as a predictor of pathogenicity of Corynebacterium group D2 which seems to act as an opportunistic pathogen in urinary tract infections.

Aged↗

Evaluation of a passive hemagglutination assay as screening test and of a recombinant immunoblot as confirmatory test for serological diagnosis of Lyme disease.

In order to evaluate a commercially available passive hemagglutination assay (PHA) as a screening test for the diagnosis of Lyme disease, 173 sera were tested by PHA and the results compared with those obtained by an indirect immunofluorescence assay (IFA) and a conventional immunoblot using whole cell antigen (IB). Identical results were found by PHA and IFA in 80% of all cases. The sensitivity of the PHA was comparable to that of the IB (96%). However, confirmation of positive PHA results was necessary due to lack of specificity. A commercially available recombinant immunoblot (RIB) was compared to a conventional IB for its efficiency as a confirmatory assay. The rate of agreement was 86% of 64 sera tested positive or negative by IB. However, in order to obtain this high concordance of the RIB and IB, it was necessary to modify the RIB interpretation criteria of the manufacturer. Thus, screening of serum specimens by the PHA and confirmation of test results by the RIB appears to be a convenient test combination that allows the serological diagnosis of Lyme disease in most cases.

Fluorescent Antibody Technique↗

Comparative evaluation of different fixation procedures and different coupling reagents for the demonstration of influenza virus-specific antibodies by the indirect hemagglutination test.

The indirect hemagglutination technique has been improved by fixing the carrier erythrocytes successively with glutaraldehyde and sulfosalicylic acid. Sensitization by covalent conjugation of influenza virus antigens to the erythrocytes with various coupling reagents, which resulted in stable and highly sensitive test cells, has been defined. An economical affinity chromatography procedure using antibody-coated agarose has been developed to prepare sufficiently pure antigens from fowl plague virus-infected choriollantoic membranes.

Animals↗

A comparative study of passive hemagglutination and immunofluorescence methods for the detection of mitochondrial antibodies in sera from patients with liver diseases.

Serum mitochondrial antibodies were capable of demonstration by the passive hemagglutination method (PHA) using sheep red blood cells coated with mitochondrial inner membrane components. Titers of mitochondrial antibodies obtained by PHA were well correlated with the antibody titers by the immunofluorescence method (IF) and the former method was more sensitive than the latter. Of 15 patients with primary biliary cirrhosis, mitochondrial antibodies were detected by PHA in the sera from 14 patients including two patients with negative results by IF. The antibodies were not detected by PHA in any sera from both 30 patients with chronic hepatitis and 26 with liver cirrhosis. In conclusion, PHA could be an useful diagnostic tool for the detection of serum mitochondrial antibodies in the patients who were suspected of primary biliary cirrhosis in spite of negative results by the conventional IF.

Antibodies↗

Detection of anti-extractable nuclear antigens in connective tissue diseases: comparison between passive hemagglutination, counterimmunoelectrophoresis and double immunodiffusion.

Antibodies to the three major components of the complex called soluble extractable nuclear antigen (ENA) were detected by passive hemagglutination (HA), counterimmunoelectrophoresis (CIE) and double immunodiffusion (DI) in 256 patients with connective tissue diseases. Anti-ENA antibodies were demonstrated by all the three employed methods in only 44.9% of the cases. These methods were not able to detect all antibodies to these antigens or any single specificity; CIE was however the most sensitive method for anti-RNP and HA for anti-Sm antibodies, while DI was the most suitable technique for serum samples with multiple anti-ENA specificities. Only in less than 50% of the cases the specificity detected by HA was comparable with that given by CIE or DI. Hence, for detecting anti-ENA antibodies a combination of these methods should be maintained, at least until more precise and reliable methods will become available.

Antibodies, Antinuclear↗