Dot immunoperoxidase assay using monoclonal antibody for direct detection of cytomegalovirus in urine samples.

A rapid, simple dot immunoperoxidase assay for the direct detection of cytomegalovirus in clinical urine samples was developed. The assay was performed on nitrocellulose paper dotted with urine pellets free of cellular debris. Cytomegalovirus was detected with a monoclonal antibody to the capsid antigen, and the complex was visualized by immunoperoxidase staining.

Stimulating effect of heat shock on the early stage of human cytomegalovirus replication cycle.

The effect of mild heat shock on the replication of human cytomegalovirus (HCMV) was studied in human embryo fibroblasts. Treatment of cell cultures at 44 degrees C for 10 min just before infection or at 24 h post infection (p.i.

Detection of specific immunoglobulin M antibodies to cytomegalovirus by using monoclonal antibody to immunoglobulin M in an indirect immunofluorescence assay.

The detection of immunoglobulin M (IgM) antibodies to cytomegalovirus-induced late antigens by an indirect immunofluorescence assay was improved by the use of monoclonal antibodies to human IgM. Nonspecific background fluorescence was absent, facilitating the reading of the slides and the detection of a specific fluorescence reaction in sera with low levels of specific IgM. Moreover, the indirect immunofluorescence assay with monoclonal antibodies to IgM proved more sensitive than the indirect immunofluorescence assay with polyclonal antibodies to IgM.

Rapid diagnosis of viral infections by an alkaline phosphatase immunocytochemical method.

The use of alkaline phosphatase immunocytochemical staining was explored for the rapid diagnosis of poliovirus, adenovirus, herpes simplex virus and cytomegalovirus infections in cell cultures. In this test, viral antigens treated with their relative antibody were incubated with alkaline phosphatase-labelled antisera. The enzyme label was developed with a naphthol salt in the presence of a diazonium salt (Fast Blue) in order to obtain a blue coloured precipitate at the site of the enzyme.

Indirect alkaline phosphatase immunoenzymatic staining for the detection of antibodies to Epstein-Barr virus-induced virus capsid antigens and early antigens.

An indirect alkaline phosphatase immunoenzymatic staining technique was developed for the detection of antibodies against Epstein-Barr virus-induced virus capsid antigens and early antigens in cell smears. The presence of antibodies against Epstein-Barr virus-induced virus capsid antigens and early antigens was revealed by a dark blue staining of cells expressing the antigens. The alkaline phosphatase assay gave a permanent record of the reaction that could be visualized under an ordinary light microscope.

Double immunoenzymatic staining for the simultaneous detection of Epstein-Barr virus induced antigens.

A double indirect immunoenzymatic staining was developed for the simultaneous visualization of Epstein-Barr virus-induced early antigens and virus capsid antigens in P3HR1 lymphoblastoid cell line. The double immunocytochemical staining was performed with a four-stage and a two-stage procedure employing human sera and monoclonal antibodies against Epstein-Barr virus-induced antigens, followed by the addition of specific alkaline phosphatase and peroxidase labeled antisera. The selection of substrates yielding reaction products of contrasting colours enabled the observer to distinguish cells expressing Epstein-Barr virus capsid antigens (blue) from cells expressing Epstein-Barr virus early antigens (brown).

Relationship between smooth muscle and cytoskeleton antibodies in neuroblastoma.

The previously reported high prevalence of smooth muscle antibodies in neuroblastoma has been found to be associated with a similarly elevated prevalence of anti-cytoskeleton antibodies. The most relevant finding is related to anti-microfilaments (anti-actin) and anti-microtubules antibodies, which were detected with highly significantly different prevalences when compared with a disease control group (p less than 0.001 and p less than 0.

Alkaline phosphatase immunoenzymatic staining for detection of antigens induced by cytomegalovirus.

An indirect alkaline phosphatase immunoenzymatic staining method was developed to localise antigens in human embryo fibroblasts that have been induced by cytomegalovirus. The enzyme label was developed with a naphthol salt and fast blue to obtain a bright blue staining of the antigens that could be clearly visualised under an ordinary light microscope. The procedure is rapid, sensitive, and specific and can be used in diagnostic laboratories to detect active infection caused by cytomegalovirus.

Detection of active Epstein-Barr infection in pregnant women.

Sera taken from pregnant patients and their newborns at delivery were examined for evidence of primary or reactivated Epstein-Barr virus infection. Of 102 women, 37 showed serological signs of reactivated and two signs of primary infection. A mild congenital defect was observed in association with one of the two cases of maternal primary infection.

Rapid neutralization assay for human cytomegalovirus antibody.

Human cytomegalovirus induces the appearance of immediate early antigens in infected cells 1 h after infection. This provided the basis for the development of a rapid neutralization assay for cytomegalovirus antibody which was able to yield results within a single day. Indirect immunofluorescence to visualize immediate early antigen-positive cells was applied to the rapid determination of cytomegalovirus-neutralizing antibody.

Relationships between smooth muscle and cytoskeleton antibodies in human serum samples.

In order to characterize the relationship between smooth muscle antibodies (SMA) and antibodies directed to the cytoskeleton components (microfilaments, intermediate filaments and microtubules), 338 serum samples from patients with various diseases and from healthy control subjects were tested for SMA and anti-cytoskeleton antibodies using a conventional immunofluorescence method employing traditional substrates and vinblastine-treated fibroblasts. The correspondence between SMA and these antibodies turned out to be incomplete and more evident for anti-microfilaments and anti-microtubules than for anti-intermediate filaments antibodies. Our data confirm that SMA are a quite heterogeneous family of antibodies, whose specificities are not entirely related to the cytoskeleton components.

Effect of heat shock on Epstein-Barr virus and cytomegalovirus expression.

The effect of heat shock was investigated on lymphoblastoid cell lines Raji and P3HR1 harbouring Epstein-Barr virus (EBV) genomes, and on Vero cells abortively infected with human cytomegalovirus (HCMV). A heat shock at 44 degrees C for 10 min induced the appearance of EBV early antigens in Raji cells and increased the percentage of cells expressing EBV viral capsid antigens in P3HR1 cells. Heat shock performed on Vero cells just before HCMV infection resulted in an approximately fourfold increase in the number of cells exhibiting early nuclear antigens, and in the appearance of HCMV-induced Fc receptors.

Serological screening for the prevention of transfusion-acquired cytomegalovirus infection.

The presence of antibodies against cytomegalovirus (CMV)-induced immediate-early antigens (IEA), early antigens (EA) and late antigens (LA) was sought in 500 healthy blood donors. Antibodies to late antigens were detected in 76% and antibodies to immediate-early and early antigens were detected in 9.6% and 10.

Rapid detection of antibodies against cytomegalovirus induced immediate early and early antigens by an enzyme linked immunosorbent assay.

An enzyme linked immunosorbent assay (ELISA) for detecting antibodies against cytomegalovirus induced immediate early antigens and early antigens was developed using purified nuclear antigens and was compared with the indirect immunofluorescence test. The tests were comparable in their ability to detect positive and negative sera, and antibody titres determined by both assays were similar. The use of ELISA for the detection of antibodies against cytomegalovirus induced immediate early and early antigens is advocated in diagnostic and research laboratories.

Brief clinical report: chondroectodermal dysplasia (Ellis-van Creveld) with anomalies of CNS and urinary tract.

We report on a 19-month-old girl with chondroectodermal dysplasia (Ellis-van Creveld) and other previously undescribed visceral and central nervous system anomalies. These anomalies include cerebral heterotopias, renal agenesis, and congenital megaureter.

Bone-metastasizing primary renal tumors in children.

Six cases of childhood renal tumor with extensive bone involvement are reported. The neoplasms had been classified originally as Wilms tumor with atypical clinical and pathologic features. Subsequent to a retrospective histologic analysis, the lesions were reclassified as follows: three cases as bone-metastasizing renal tumors of childhood, one case as rhabdomyosarcoma, one case as undifferentiated sarcoma, and one case as undifferentiated malignant neoplasm.

Can Epstein-Barr virus infect and transform all the B-lymphocytes of human cord blood?

Quantitative aspects of Epstein-Barr virus infection and transformation of human neonatal B-lymphocytes have been investigated. 72 to 90% B-cells were obtained with enrichment. Of the B-cells, 19 to 97% showed nuclear antigen (EBNA) 2 days after infection.

Influence of cell cycle on the efficiency of transfection with purified human cytomegalovirus DNA.

The efficiency of transfection of purified HCMV genome was studied in synchronized human embryo fibroblasts. The data show that HCMV DNA infection performed in S phase is more efficient than in cells infected in G2, M and G1 phases of cell cycle, respectively. These findings stress the importance of the metabolic state of cell cultures in HCMV genome transfection.

Human cytomegalovirus cycle in synchronized cells: electron microscope study.

Human Cytomegalovirus (HCMV) was inoculated in synchronized human fibroblasts at different phases of the cell cycle. the virus replication appeared strongly dependent on the host metabolic state, being faster when the infection was carried out in G2 and S phase.

Serum antibody profile against herpesviruses in renal transplant recipients.

Forty-eight renal allograft recipients who had received a kidney graft 2 months to 8 years previously, were followed for the presence of serum antibody against Cytomegalovirus (CMV), Epstein-Barr virus (EBV), Herpes simplex virus (HSV), and Varicella-Zoster virus (VZV). These investigations were performed over a 6 months period and in at least two serum samples from each patient, obtained at intervals of 2-3 months. The presence of serological signs of recent or active infection were observed in 31 patients for CMV, in 22 patients for EBV (18 patients were positive both for CMV and EBV) and only in 3 patients for HSV, while no patients showed serological signs of recent or active infection by VZV.

Inhibition of a complete replication cycle of human cytomegalovirus in actinomycin pre-treated cells.

The study of human cytomegalovirus (HCMV) in cultures of human embryo lung fibroblasts, pre-treated with actinomycin D, has shown that under these conditions the virus infection does not proceed beyond the 'early' events of the virus replication cycle. In the same experimental conditions the growth of poliovirus type I, vaccinia virus and herpes simplex type I virus, was completely unaffected. These results suggest that the complete HCMV replication cycle requires some cellular function(s) between early transcription of the input virus genome and virus DNA synthesis.

Cytomegaloviru; antibody in the urine of renal transplant patients.

Small amounts of cytomegalovirus (CMV) antibody were detected in the urine of renal transplant patients excreting the virus. The antibody was probably produced locally, as a result of active CMV infection of the urinary tract.