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Comprehensive Viral Detection and Profiling of Plasma Cell-Free RNA in Patients With Suspected Hemophagocytic Lymphohistiocytosis.

Hemophagocytic lymphohistiocytosis (HLH) is a severe, rapidly progressive disease. While viral infection is considered a common etiology of pediatric HLH, specific causative viruses other than the Epstein-Barr virus (EBV) have been rarely identified. This study utilized metagenomic next-generation sequencing (NGS) to identify potential causative pathogens in plasma samples from 17 pediatric patients with suspected HLH. Additionally, one case each of confirmed EBV- and cytomegalovirus (CMV)-associated HLH was analyzed for methodological validation. Plasma cell-free RNA (cfRNA) profiling was performed using NGS data to assess the host transcriptome response. Significant viral reads of human herpesvirus-6B, human herpesvirus-7, and Hubei reo-like virus (HRLV) 14 were detected using metagenomic NGS in one patient each. Plasma cfRNA profiles from five patients with viral infection (including EBV and CMV) were compared to those of 14 patients without viral infection. By comparing the two patient groups, 1053 differentially expressed genes were identified. The gene ontology (GO) term of "adaptive immune response" (GO: 0002250) was significantly enriched among upregulated genes in the virus-positive group. Furthermore, an isolated cluster consisting specifically of mitochondrial RNAs, was identified in the upregulated genes of the virus-positive group. Using metagenomic NGS, several candidate viral pathogens were identified in patients with suspected infection-related HLH. The viral genome of HRLV 14, previously undetected in human clinical samples, was identified in one patient. The results from plasma cfRNA profiling suggest that mitochondrial RNAs may reflect the underlying pathogenesis of virus-associated HLH and have potential utility as disease biomarkers.

Humans

In vitro-labeled DNA for detecting viral genomes in multiple sclerosis: I. Papovaviruses.

Papovaviruses appear to be neurotropic and one, JC virus, is implicated as the cause of one type of demyelinating disease, progressive multifocal leukoencephalopathy. To investigate whether human papovaviruses play a role in multiple sclerosis, radioactively labeled DNA from BK virus, human papilloma virus, and simian virus 40 was used as a probe in order to detect related unlabeled DNA sequences in DNA isolated from multiple sclerosis brain and/or spinal cord. Labeled viral probes were denatured and DNA allowed to reassociate in the presence of excess unlabeled DNA from multiple sclerosis tissue or from controls. The reassociation rate of the probe is proportional to the concentration of viral DNA present, and an increase in the reassociation rate of the probe over that of control reactions would indicate the presence of unlabeled viral DNA in multiple sclerosis cellular DNA. However, addition of DNA derived from multiple sclerosis patients did not increase rates of reassociation of viral probes. Known human papovaviruses probably have no role in the pathogenesis of multiple sclerosis.

BK Virus

[Experience in examining infectious hepatitis patients by using a viral detection method].

Immune electron microscopy was shown to be an acceptable diagnostic method which allowed to detect hepatitis A virus particles in the stools of patients with infectious hepatitis even at the late stages of the disease. The examination of stools from 225 children (aged 2--17 years) hospitalized with diagnosed infectious hepatitis during the period of a seasonal rise in morbidity rate yielded positive findings at an average rate of 10.5%. This rate also depended on the severity of the disease and the patient's age. The results obtained indicated that patients with infectious hepatitis could shed the virus for a long time (about 2 weeks) after the onset of the disease. Patients with the atypical, or anicteric, form of the disease could be equally considered a source of virus shedding. The prolonged excretion of viral particles did not correlate with delayed IgM response. The particles identified as hepatitis A virus seemed to be antigenically very close, if not identical, to the virus circulating in some other countries (USA, Australia, West Germany).

Adolescent

Detection of viral proteins in mouse mammary tumors by immunoperoxidase staining of paraffin sections.

An indirect immunoperoxidase method is described, which can readily detect viral antigens in paraffin sections of primary, transplanted, and metastatic mammary tumors of mice. In addition to having the obvious advantage of not being limited to fresh specimens, immunoperoxidase staining of paraffin sections proved to be superior in many respects when compared with immunofluorescence and frozen sections. Immunoperoxidase staining of paraffin sections is permanent and provides the kind of histological detail required for precise cytological identification and localization with light microscopy. All of 25 tumors and 4 metastatic lesions showed evidence of glycoprotein gp52 as well as other mouse mammary tumor viral antigens. The pattern and intensity of the stain were related to the degree of histologic differentiation of the tumor. Wide variations in expression of viral antigens by individual malignant cells were observed within the same tumor.

Animals

Sensitivity of radionuclide brain scan and computed tomography in early detection of viral meningoencephalitis.

The sensitivity of radionuclide imaging and computed tomography (CT) was evaluated in 25 patients for early detection of viral meningoencephalitis. Diagnosis was based on clinical evidence, cerebrospinal fluid (CSF) studies, electroencephalography (EEG) and radionuclide imaging. Computed tomography with contrast enhancement was performed within four days after onset of neurological signs or symptoms in 23 patients; no significant findings such as low-absorption abnormalities, mass effect or abnormal enhancement were seen. Radionuclide imaging demonstrated a sensitivity of 90% in the detection of viral meningoencephalitis; the temporal lobe was most commonly involved in patients with herpes encephalitis. Radionuclide imaging should be considered as the first diagnostic procedure in suspected early viral meningoencephalitis.

Adolescent

Biochemical approaches to detection of viral related information in human acute leukemic cells.

Two biochemical markers that have been utilized for the detection of viral related information in human acute leukemic cells are: 1) The reverse transcriptase and 2) a high molecular weight RNA with viral related nucleotide sequences. This paper summarizes evidence that shows that the reverse transcriptase isolated from human acute leukemic cells is biochemically related to the reverse transcriptase from RNA tumor viruses.

Acute Disease

Detection of viral hepatitis type A.

Several technics are currently being used to detect hepatitis type A antigen or its antibody. These include immunoelectronmicroscopy, immune adherence, and complement fixation. This paper describes another promising technic, a microtiter solid-phase immunoradiometric assay, in which hepatitis A antigen and antibody are detected. Such a method can be utilized for biochemical and biophysical analysis of purified particles, for the seroepidemiology of type A hepatitis, and as a means for monitoring hepatitis A antigen in cell cultures.

Animals

Metaproteomic profiling reveals viral proteins and associated host proteomic alterations in glioblastoma.

Glioblastoma (GB) is a WHO grade 4 brain cancer with dismal prognosis, yet its aetiology remains poorly defined. Although viral involvement has been proposed, findings across studies remain inconsistent, reflecting inherent limitations of individual technologies and cohort size. Here we applied metaproteomic profiling to a publicly available GB proteome dataset (12 control, 21 adjacent, 159 tumour) and an independent cohort of 81 samples (37 control, 44 tumour) to detect viral proteins in tumour and controls tissues. Across cohorts, we detected viral proteins from diverse species, with human herpesviruses (HHV-1, 2, and 8) more frequently detected in GB tumours compared with control tissues. Analysis of the host tumour proteome revealed differential abundance of proteins related to transcriptional regulation, RNA processing, protein translation, immune responses, and mitochondrial-associated metabolism. Correlation analysis identified associations between viral and human proteins, with several linked to biological processes previously implicated in DNA virus-host interactions. Further stratification of tumour by HHV-1 status showed consistent alterations in proteins associated with mitochondrial-associated metabolism, protein turnover, and cell adhesion/signalling.In summary, this study demonstrates the feasibility of metaproteomics for detecting viral components in archival GB tissues. Using this approach, we observed differences in viral protein landscape across cohorts and identified associations between viral presence and host proteomic features, providing a protein-level framework for future studies of virus-host interactions in GB.

Humans

Identification of murine leukemia viral antigens detected on mouse cells by H-2 alloantisera.

H-2 alloantisera have been previously reported to contain antibodies against murine leukemia viral antigens, but the nature of the viral antigens on mouse cells which interact with these antibodies has not been established. We have found that H-2 alloantisera recognize components of molecular weight 70 000-80 000 mouse lymphocytes and leukemia cells. These components were also detected by a goat antiserum against the murine leukemia virus (MuLV) glycoprotein (gp 70) and are therefore closely related to or identical with that viral protein. Although most H-2 alloantisera detected gp 70-like molecules on lymphocytes and leukemia cells from a great variety of mouse strains, only one H-2 alloantiserum was found to interact with a gp 70 component on cells from C57BL/10 and C57BL/6 mice. Animals such as C57BL/10 mice that lacked the component reacting with most H-2 alloantisera showed increased serum levels of anti-MuLV antibodies after injection of B10.A spleen cells having a gp 70 component detectable by other H-2 alloantisera. In contrast, strains with cells reactive to antiviral antibodies in the H-2 alloantisera had low responses to MuLV antigens after a similar immunization procedure. Serum levels of anti-MuLV antibodies in both groups of mice, however, were increased after injection of Freund's adjuvant. These observations suggest that anti-MuLV antibodies in mouse alloantisera may arise from a response to viral antigens on the immunizing cells and general stimulation of the immune system.

Animals

Viral surveillance beyond detection: JMTV and the need for ensemble approaches in emerging virus discovery.

The recent report by T. Murillo, L. E. Enrique Chaves-González, S. Temmam, S. Bermúdez, et al. (Microbiol Spectr 14:e04078-25, 2026, https://doi.org/10.1128/spectrum.04078-25) expands the known geographic and ecological range of Jingmen tick virus (JMTV) by detecting the virus in Amblyomma mixtum ticks collected from horses in Costa Rica. This is an important finding because A. mixtum can feed on wildlife, domestic animals, and humans, creating a possible interface for virus movement across various hosts. The study also places the Costa Rican virus in a wider phylogenetic context, linking it to JMTV diversity reported from other regions. However, the detection of viral RNA in ticks should not be interpreted as proof of local disease, human infection, or active transmission, especially in the absence of supporting results. Instead, it reflects an important signal for careful viral surveillance. Here, I discuss how JMTV illustrates the need for ensemble approaches that combine field sampling, phylogeny, segment-level genome analysis, serology, experimental validation, and data-driven virus discovery tools.

emerging viruses

An immunofluorescence diagnostic test for avian infectious laryngotracheitis.

An antiserum to a recent field isolate of infectious laryngotracheitis virus was conjugated with fluorescein isothiocyanate and used to detect viral antigen to infected chorioallantoic membranes and trachael epithelium by the direct fluorescent antibody technique. In experimentally infected birds, viral antigen was detected with the fluorescent antibody technique from 2 to 14 days post-inoculation but histological evidence of tracheitis was only observed from day three to day ten. The fluorescent antibody test detected 22 of 23 histologically confirmed cases of ILT and was more accurate than virus isolation when used in the diagnosis of respiratory disease from field outbreaks. It was concluded that the speed, accuracy and sensitivity of the fluorescent antibody technique make it a useful tool in the diagnosis of infectious laryngotracheitis.

Animals

Detection of viral sequences of low reiteration frequency by in situ hybridization.

The sensitivity of in situ hybridization has been increased at least 10-fold by hybridizing in cDNA excess, by increasing the diffusion of the cDNA through the cells, by hybridizing at optimum temperature, and by stabilizing hybrids during autoradiography. Saturation of intracellular RNA with [3H]cDNA has been achieved. The assay is quantitative. In situ hybridization has been used to detect and quantitate visna virus RNA in infected cells. By using [3H]cDNA with specific activity of 2 X 10(8) dpm/micrograms and conditions that reduce background to negligible levels, 10--20 copies of viral RNA per cell can be detected and quantitated after 2 days of autoradiographic exposure.

Animals

Biological characteristics of a stimulatory factor for viral replication detected in egg fluids.

Yolk sac and allantoic fluids and albumen from uninfected chicken eggs contain a low-molecular weight factor which, after 90 minutes of contact with cell cultures, significantly enhances viral replication. Of several viruses tested, Semliki Forest virus exhibited the highest (10(5)-fold) responsiveness to this enhancing factor. Maximal enhancement was obtained with cells subjected to low multiplicities of infection. The stimulation of viral replication was linear with the age of the cells in culture. The enhancing factor did not operate through an anti-interferon mechanism. However, it exerted a boosting effect on the low-grade cellular metabolism of ageing cell monolayers. Some of the physico-chemical features of the enhancing factor were determined and conjectures concerning its chemical makeup are discussed.

Allantois

Type C oncornavirus isolation studies in systemic lupus erythematosus. II. Attempted detection by viral RNA-dependent DNA polymerase assay.

Isolation of type C oncornavirus was attempted from 20 tissues and cell cultures of patients with systemic lupus erythematosus. Chemical inducers, cocultivation and fusion with cells from multiple other species, prolonged subculturing, and the RNA-dependent DNA polymerase assay for virus detection were used. A type C virus was isolated, but was shown to be the endogenous rat virus. Thus the methods, although generally appropriate, were not specifically permissive for replication of a human type C virus. This agrees with the failure of other investigators to isolate a virus of undisputed human origin. Combining available evidence, a fundamental role for type C viruses in lupus erythematosus remains an attractive hypothesis.

Adolescent