Solid-phase enzyme-immunoassay for the detection of herpes simplex virus antigens in clinical specimens.

An indirect solid-phase enzyme-immunoassay (EIA) for the detection of herpes simplex virus (HSV) antigens in clinical specimens was developed. Rabbits and guinea pigs were hyperimmunized with highly purified nucleocapsids of HSV type 1. Microtitre plates were coated with 0.25 microgram of guinea pig anti-herpes simplex type 1 immunoglobulins per well. Clinical specimens, diluted in phosphate buffered saline containing fetal calf serum and detergents, were sonicated and incubated in the test wells overnight at 37 degrees C. Rabbit anti-HSV immunoglobulins were added as a secondary antibody at a concentration of 3.2 micrograms per well, and peroxidase conjugated swine antibodies against rabbit immunoglobulins, diluted 1:1,000, were used as a fourth layer. Clinical specimens which were sent for virus isolation or for isolation of Chlamydia trachomatis were tested by the developed assay and 20 out of 27 isolation positive specimens were found positive by EIA. Five out of 67 specimens negative by isolation gave positive results by EIA. The specificity of the results was confirmed by a control test using wells coated with normal guinea pig immunoglobulins. The test detected antigens from both serotypes of HSV. Cross reactions with varicella-zoster- or with cytomegalovirus were not found, and antigens from uninfected cells did not result in false positive results.

A "reverse" solid-phase radio-immuno-assay for IgM-antibodies to hepatitis A virus.

A "reverse"solid-phase radio-immuno-assay for IgM antibodies to hepatitis A virus (HAV) was developed. Anti-human IgM immunoglobulins were bound on the wells of polyvinylchloride microtiter plates. Serum specimens were incubated in the anti-human IgM coated wells and bound IgM antibodies were then assayed for antigen specificity by subsequent incubations with HAV antigen and 125I-labelled human anti-HAV IgG. The test showed a high sensitivity and specificity for anti-HAV IgM antibodies. No false-positive reactions were observed either in the sera from patients with hepatobiliary disorders other than HAV infection or in the sera containing both rheumatoid factor and anti-HAV IgG antibodies. In acute HAV infections specific IgM antibodies were present already in the first specimens taken within a few days after the onset of jaundice. The persistence of the IgM antibodies was from 4 to 6 months. IgM antibody titers up to 1,000,000 were observed in the acute phase of HAV infection. In routine diagnostic work the titration of the sea was not necessary, since a reliable qualitative result was obtained by testing the sera in a single dilution of 1:100. A similar "reverse" immuno-assay principle may be adaptable for the diagnostic determination of IgM antibodies to different viral and microbial antigens.

Solid-phase enzyme immunoassay for determination of antibodies to cytomegalovirus.

A solid-phase enzyme immunoassay for the determination of immunoglobulin H (IgG) and IgM antibodies to cytomegalovirus is described. The enzyme immunoassay gave reliable and consistent results which were in concordance with those obtained by the complement fixation test and the indirect immunofluorescence test. Antibodies to herpes simplex and varicella-zoster viruses did not interfere in the enzyme immunoassay for cytomegalovirus IgM antibodies. In a few patients with IgM antibodies to Epstein-Barr virus, cytomegalovirus IgM antibodies were also detected. False-positive cytomegalovirus IgM antibody results were observed in sera containing both the rheumatoid factor and cytomegalovirus IgG antibodies. This rheumatoid factor interference was overcome by the absorption of sera with polymerized human gamma globulin. The absorption did not affect true cytomegalovirus IgM antibody titers. Also described is a simple enzyme immunoassay that makes possible a more sensitive detection of the rheumatoid factor than the latex agglutination test.

[Ultrasound testing in the diagnosis and management of maxillary sinusitis in children (author's transl)]. / Die Verlaufskontrolle des Heilungsprozesses der kindlichen Kieferhöhlenentzündung mit der Ultraschalldiagnostik.

A- and B-mode ultrasound testing was used in conjunction with X-ray studies as screening tests for the detection of maxillary sinusitis in 280 children (aged 4-10 years) about to undergo adenoidectomy, and/or tonsillectomy and/or tympanostomy. X-rays were abnormal in 114 (41%) children, while sinus disease with the retention of discharge was diagnosed by ultrasound in 63 (23%) children. These latter children were then treated for ten days with amoxicillin and decongestant (Rinexin). At time of initial surgery, 63 maxillary sinuses (in 43 of the children) were punctured and irrigated. Another group (consisting of 27 sinuses in 20 children) was treated without antral puncture. All children were reexamined after 10, 20 and 30 days with ultrasound and radiography. After 10 days, there was no discharge found in 86% of the sinuses treated by antral puncture and in 85% of the control sinuses. After one month, all sinuses which had been irrigated were found to be free of disease, while three (11%) of the control group contained discharge. However, the healing process in all patients seemed to be slower, as evidenced by sequential sinus X-rays.--We believe that ultrasonic evaluation of the maxillary sinuses is a simple, rapid and reliable method for the diagnosis and followup of maxillary sinusitis in children. With this technique, it is possible to follow the disappearance of residual sinus discharge as well as diagnose cases which need active treatment.

Local IgA-class antibodies against respiratory viruses in middle ear and nasopharyngeal secretions of children with secretory otitis media.

The presence of antibodies against some important respiratory viruses in the middle ear and nasopharyngeal secretions of 52 children with secretory otitis media (SOM) was investigated in order to find out about the role of these viruses in the development of SOM. The method employed was a sensitive radioimmunoassay test. Antibodies to adeno, syncytial, and parainfluenza 3 viruses were detected in about 50%, 50% and 20% of the patients, respectively. In eight patients the middle ear secretion/nasopharyngeal secretion titer ratio of antibodies to one virus was selectively increased in comparison with the other viruses tested, indicating an active local production of specific antibodies in the middle ear. Further studies are required to determine the cause of such an active antibody synthesis and its possible role in the etiopathogenesis of SOM.

[Otosclerosis in children (author's transl.)]. / Otosklerose im Kindesalter

11 children (13 ears) have been operated upon for otosclerosis. In ten cases the stapes footplate was found totally fixed, with histological verification of the diagnosis of otosclerosis. Conservative surgery of the oval window was possible in only four cases. All other patients required total removal of staples or footplate. Findings indicate that clinical otosclerosis occurring in childhood results in an otosclerotic process in the oval window region with total fixation of the footplate. Thus, correction usually requires removal of at least the entire footplate.

Hemagglutination by BK virus, a tentative new member of the papovavirus group.

Some characteristics of hemagglutination (HA) by the BK virus, a new candidate for the papovavirus group, have been studied. Hemagglutinin prepared from cell cultures was found to be partially masked by inhibitors which could be dissociated from the virus by incubation at 37 C or by fluorocarbon extraction. Optimal conditions for HA are outlined. In routine tests, 0.5% human erythrocytes were used. The reaction was carried out at pH 7.0 on ice-water slurry. BK hemagglutinin receptors on human erythrocytes were found to be more resistant to neuraminidase than polyoma receptors. By gradient centrifugation analysis, two types of particles were found to be responsible for HA: (i) full, deoxyribonucleic acid-containing particles with a density of 1.325 g/cm(3) and (ii) empty capsids with a density 1.29 g/cm(3). Based on particle counting, one HA unit was calculated to correspond to 3 x 10(6) virus particles.