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At least 91 records · Page 5Linked to original sources
PubMed2006

Respiratory syncytial virus detection by Remel Xpect, Binax Now RSV, direct immunofluorescent staining, and tissue culture.

The performance characteristics of Xpect RSV (XP) and Binax Now RSV (BN) were compared to those of direct fluorescent-antibody staining and/or tissue culture for detection of respiratory syncytial virus (RSV) in nasopharyngeal aspirate and wash samples from children (n = 110) and adults (n = 66). The sensitivity, specificity, positive predictive value, and negative predictive value of XP were 75%, 98%, 95%, and 90%, respectively; and those of BN were 74%, 100%, 100%, and 90%, respectively. The performances of the assays were similar within a given age group and specimen type (nasopharyngeal aspirate or wash specimen). XP and BN are useful for screening for RSV in respiratory specimens when large volumes are tested or low levels of staffing occur.

Adolescent↗
PubMed2002

Type-specific antibody for hepatitis C virus detected by use of NS-4 peptide and hepatitis C virus genome in Korea.

Hepatitis C virus, often possessing mutant genes, have the features that allow them to avoid host's immunologic response and further cause chronic progressive infections. Therefore, it is essential for those patients infected of HCV to receive improved diagnostic procedures. And it is equally important to investigate the course of disease progression and the response to treatment. The goal of this study is to review the efficacy of the third generation immunoblot assay and standardized RT-PCR-Hybridization assay, and in contrast with genotype identification(genotyping), the followings were briefly evaluated for the efficacy of serotype identification(serotyping) by using NS-4 peptide in the observation of the course and in the treatment of patients with HCV hepatitis. 1. The true positive rate in 132 cases showing repeated positives with 3rd generation anti-HCV EIA are 81.8% by immunoblot assay and 75.8% by RT-PCR-Hybridization assay. 2. The 79.5% concordance of immunoblot and RT-PCR-Hybridization assay is shown. The negative results from immunoblot assay are also negative in RT-PCR-Hybridization assay. 3. Among 95 patients with HCV hepatitis patients in 95 cases, the serotype 1, 2 and 4 were 53.2%, 45.2%, and 1.6%, respectively. In 29 cases, the genotypes of patients with HCV showed 1b in 15 cases, 2a/2c in 8 cases, 2b in 2 cases and mixed type in 4 cases. 4. In comparison between serotype and genotype, they showed 75.9% concordance. But serotyping showed higher efficacy in experimental procedures and sampling conditions, with more convenience. Based on above evaluation and reference review, it is reasonable to check with 3rd generation immunoblot assay the samples producing repeated positive results from anti-HCV EIA. For more definitive diagnosis of HCV infection, it is appropriate to confirm and double-check with standardized RT-PCR-Hybridization assay. Lastly, it is strongly suggested that for observation of progression and for choice of interferon treatment, serotype identification(serotyping) is more useful in practice than genotype identification (genotyping).

Genome, Viral↗
PubMed2005

[Oligonucleotide microarray for human immunodeficiency virus detection].

OBJECTIVE: To develop an oligonucleotide microarray for fast detection of human immunodeficiency virus (HIV). METHODS: With complete genome sequence of HIV-1 subtype B (U26942) as the target sequence and bioinformatics software such as DNAClub, Oligo6.0, BLAST, Alignment, oligonucleotide probes of high specificity with identical length and similar melting temperature (T(m)) were designed and synthesized. Oligonucleotide microarray was prepared using Cartesian Microarrayer. Using the plasmids of HIV-1 subtype B(U26942), C(U46016), F(AF075703), G(AF061640) and restriction display technique, Cy3-labeled HIV DNA fragments were amplified and the hybridization results were scanned and analyzed with Array-Pro. RESULTS AND CONCLUSION: Twenty-two optimized oligonucleotide microarray probes were obtained and used to prepare the oligonucleotide microarray for further screening studies. The microarray prepared significantly enhanced the sensitivity, reliability and speed of DNA assay, and possesses the potential for application in clinical setting.

Base Sequence↗
PubMed1974

Development of methods for detecting viruses in solid waste landfill leachates.

Methods were developed for detecting and concentrating enteric viruses in municipal solid waste landfill leachates. Poliovirus added to a leachate was not readily detectable, possibly because the virus was adsorbed to the leachate particulates. The masking effects associated with suspended solids in the leachate were overcome by adding a final 0.1 M sodium (tetra)ethylenediaminetetraacetate concentration to the leachate. A sodium (tetra)ethylenediaminetetraacetate-treated leachate could be clarified by filtration at pH 8.0 without a loss of virus. The clarified and sodium (tetra)ethylenediaminetetraacetate-treated leachate contained interfering materials of an anionic nature which prevented virus adsorption to epoxy-fiber glass filters. This interfering effect was overcome by treating the leachate with an anion-exchange resin. Viruses in the resin-treated leachate were concentrated by adjusting the leachate to pH 3.5, adding AlCl(3) to a final 0.005 M concentration, adsorbing the viruses to an epoxy-fiber glass virus adsorbent, and eluting the adsorbed viruses in a small volume. When this method was used to concentrate poliovirus 100-fold in a variety of leachates, the average virus recovery efficiency was 37%. With the methods described in this study, it should be possible to efficiently monitor solid waste disposal site leachates for enteric viruses.

Adsorption↗
PubMed2003

Cloning and nucleotide analysis of segment A gene of infectious bursal disease virus detected in Korea.

A strain of infectious bursal disease virus (IBDV) was detected from bursal tissues of chicks which suffered from infectious bursal disease (IBD) in Chinju, Korea and provisionally named as Chinju strain. A full-length cDNA clone for segment A gene of the virus was constructed, and complete nucleotide sequence of the gene including noncoding region was determined and analyzed by comparison with that of other IBDV strains. The segment A gene of Chinju strain consisted of 3,269 nucleotides including 862 adenine (26.4%), 917 cytosine (28.0%), 854 guanine (26.1%) and 636 thymine (19.5%). There were regions for two open reading frames (ORFs), ORF1 encoding the VP5 with ATG codon at nucleotides 98-100 and ORF2 encoding the polyprotein of VP2, VP4 and VP3 in the nucleotides 132-3,170. In deduced translation the ORF2 encoded 1,012 amino acids. The full nucleotide sequence of segment A gene and amino acid sequence of ORF2 of the Chinju strain showed 98-99% homology with those of the very virulent IBDVs (vvIBDVs) such as HK46, OKYM, UK661, UPM97/ 61, D6948 and BD3/99. In phylogenetic analysis of nucleotide and amino acid sequences, the Chinju strain was also related closely to the vvIBDVs. Hence, it was suggested that the Chinju strain is a vvIBDV. The nucleotide and amino acid sequences of the Chinju strain with pertinent information can be useful for the development of genetically engineered vaccines and diagnostic reagents against vvIBDV.

Amino Acid Sequence↗
PubMed2002

Viruses detected in the caecum contents of healthy pigs representing a new genetic cluster in genogroup II of the genus "Norwalk-like viruses".

Viruses of the genus 'Norwalk-like viruses' (NLVs) detected in humans have been genetically classified into two major genetic groups, genogroups I and II (GI and GII), which together are made up of at least 14 genetic subgroups. However, a comparable classification of NLVs in other species remains to be carried out. We sequenced a 2-kb region from within the RNA polymerase gene to the 3' end of open reading frame 2 (ORF2) of two NLV strains previously detected in the caecum contents of healthy pigs. The sequences of the entire ORF2 of these two NLV strains were analyzed for their genetic relationships to 15 human strains, which have already been reported and used as references for the genetic classification of human NLV strains, and additional two strains; one, a human strain which has recently been reported and appears to represent a new genetic subgroup of GII; and the other, an animal NLV strain. Analysis of a matrix showing pairwise identities and topology of a neighbor-joining tree showed that the two swine strains could be classified into a new genetic subgroup of GII on the basis of the amino acid sequences of the entire capsid protein. Grouping of the two swine strains was well corroborated by results of similar analyses of nucleotide sequences of the entire ORF2 and of a 510 base region at the 3' end of ORF1.

Animals↗
PubMed1992

Variation in the biological function of envelope protein epitopes of yellow fever vaccine viruses detected with monoclonal antibodies.

Three different virus strains (17D-204, 17DD and the French neurotropic vaccine) have been used as live attenuated yellow fever (YF) vaccines and are manufactured in different centres around the world. The envelope proteins of these vaccine viruses were examined and compared using mouse monoclonal antibodies (MAbs) in haemagglutination inhibition (HAI) and neutralization (N) tests. The epitopes eliciting HAI and/or N were found to vary depending on the virus examined. Such variation was also found between vaccine viruses of the same strain manufactured in different centres. These data were confirmed by the use of mouse polyclonal antisera. On the basis of the MAb results in HAI tests a dendrogram of the similarity coefficients between the viruses was constructed and showed that the viruses could be placed into three major groups. Thus, it is concluded that YF vaccines manufactured in different centres are antigenically distinct as recognized by the mouse immune system.

Antibodies, Monoclonal↗
PubMed2005

[Preliminary study on genotype of hepatitis B virus detected from Tibetans in China].

OBJECTIVE: To determine the main genotype of hepatitis B virus (HBV) detected from Tibetans in China and provide basic data for hepatitis control and prevention. METHODS: The S gene and C gene were amplified by PCR from the sera of HBsAg positive Tibetans. After sequencing, the gene sequences were analyzed and the phylogenetic trees were drawn by the software MEGA3. RESULTS: In trees based on S gene, the sequences of most samples clustered at genotype D, while in trees based on C gene, the sequences of all samples clustered at genotype C. CONCLUSION: The dominant genotype of HBV detected from Tibetans in China is a C/D hybrid.

Genotype↗
PubMed1998

Aujeszky's disease (pseudorabies) virus detection in cerebrospinal fluid in experimentally infected pigs.

The presence of Aujeszky's disease virus in cerebrospinal fluid of experimentally infected pigs was studied using the techniques of virus isolation and PCR. Pigs, some of which were previously vaccinated against Aujeszky's disease, were inoculated with different doses of the Aujeszky's disease NIA-3 strain. At the time of death or sacrifice, a sample of cerebrospinal fluid was taken and tested for the presence of virus using the mentioned techniques. Virus was isolated only from one sample, while it was detected by PCR in most of them. The higher sensitivity of the PCR technique and the possible presence of antiviral antibodies in the cerebrospinal fluid are reasons that can be argued to explain this fact. By PCR, the virus was detected more efficiently when digested cerebrospinal fluid cells were used as DNA source than when using whole cerebrospinal fluid, suggesting that the virus could be cell-associated. Aujeszky's disease virus could not be detected by PCR in pigs which survived the acute phase of the infection and were euthanased at 8 weeks post-inoculation, when they were latently infected. This indicated that the cerebrospinal fluid is not an adequate sample for the diagnosis of latency. Since Aujeszky's disease virus was detected from most of the tested samples, we believe that this could be an adequate procedure for the quick diagnosis of Aujeszky's disease.

Animals↗
PubMed1980

Further studies on the use of protein A in immune electron microscopy for detecting virus particles.

The immune electron microscopic technique which involves pre-coating electron microscope grids with protein A before coating them with the specific antiserum, has been found suitable for detecting isometric insect and plant viruses. With the three virus-antibody combinations tested, the optimum antiserum dilution for protein A plus antiserum treatment was found to be 1:100 or less, whereas in the case of grids treated with antiserum alone it ranged from 1:1000 to 1:2000 although the titre of the antisera ranged from 1:512 to 1:4096. The increase in the number of particles on grids treated with protein A plus antiserum over those treated with antiserum alone, at each optimal antiserum dilution, was 25.7-fold (sugarcane mosaic virus), 2.1-fold (tobacco mosaic virus) and 1.6- and 2.4-fold (Erysimum latent virus--two different sap dilutions). Protein A plus antiserum-coated grids can be stored for up to 6 months at 4 degrees C, but not at room temperature or in a desiccator, and still retain about 25% of their activity, sufficient to detect any virus using the electron microscope. Antisera preserved in glycerol can be used successfully for detecting viruses by immune electron microscopy.

Immunologic Techniques↗
PubMed2005

Multiplexed flow cytometric immunoassay for influenza virus detection and differentiation.

Microsphere-based immunoassay by flow cytometry has gained popularity lately in protein detection and infectious disease diagnosis due to its capacity for multiplexed analysis and simple assay format. Here, we demonstrated the power of microsphere-based immunoassay for high-sensitivity detection and accurate differentiation of influenza viruses. The effects of sample volume and bead number on the assay sensitivity of viral antigen detection were studied. Compared to enzyme-linked immunosorbent assays, flow-based bead assays provided approximately 10-fold lower detection limit for viral particle detection and performed similarly for recombinant viral hemagglutinin protein detection. A four-plexed assay for influenza virus typing and influenza B virus sublineage characterization was developed to demonstrate the potential for multiplexed viral antigen detection and differentiation.

Flow Cytometry↗
PubMed2005

West Nile virus detection in urine.

We report West Nile virus (WNV) RNA in urine collected from a patient with encephalitis 8 days after symptom onset. Viral RNA was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Sequence and phylogenetic analysis confirmed the PCR product to have > or = 99% similarity to the WNV strain NY 2000-crow3356.

Aged↗
PubMed1972

Virus-specific ribonucleic acid in cells producing rous sarcoma virus: detection and characterization.

Cells producing Rous sarcoma virus contain virus-specific ribonucleic acid (RNA) which can be identified by hybridization to single-stranded deoxyribonucleic acid (DNA) synthesized with RNA-directed DNA polymerase. Hybridization was detected by either fractionation on hydroxyapatite or hydrolysis with single strand-specific nucleases. Similar results were obtained with both procedures. The hybrids formed between enzymatically synthesized DNA and viral RNA have a high order of thermal stability, with only minor evidence of mismatched nucleotide sequences. Virus-specific RNA is present in both nuclei and cytoplasm of infected cells. This RNA is remarkably heterogeneous in size, including molecules which are probably restricted to the nucleus and which sediment in their native state more rapidly than the viral genome. The nature of the RNA found in cytoplasmic fractions varies from preparation to preparation, but heterogeneous RNA (ca. 4-50S), smaller than the viral genome, is always present in substantial amounts.

Aspergillus↗
PubMed1975

Serological analysis of an oncornavirus (PMF virus) detected in malignant permanent human cell lines.

Immunodiffusion analysis of the PMF virus which was detected in malignant permanent human cell lines revealed positive reactions with antisera against the Mason-Pfizer monkey virus (MPMV). No cross-reactivity was demonstrated with murine leukemia virus (MuLV), rat leukemia virus (RaLV), hamster leukemia virus (HaLV), feline leukemia virus (FeLV), simian (woolly monkey) sarcoma virus (SSV-1) and mouse mammary tumor virus (MTV). The cross-reactive antigens of the PMF virus and the MPMV are considered as evidence for the human origin of the PMF virus.

Animals↗
PubMed2004

Monkeypox virus detection in rodents using real-time 3'-minor groove binder TaqMan assays on the Roche LightCycler.

During the summer of 2003, an outbreak of human monkeypox occurred in the Midwest region of the United States. In all, 52 rodents suspected of being infected with monkeypox virus were collected from an exotic pet dealer and from private homes. The rodents were euthanized and submitted for testing to the United States Army Medical Research Institute of Infectious Diseases by the Galesburg Animal Disease Laboratory, Illinois Department of Agriculture. The rodent tissue samples were appropriately processed and then tested by using an integrated approach involving real-time polymerase chain reaction (PCR) assays, an antigen-detection immunoassay, and virus culture. We designed and extensively tested two specific real-time PCR assays for rapidly detecting monkeypox virus DNA using the Vaccinia virus F3L and N3R genes as targets. The assays were validated against panels of orthopox viral and miscellaneous bacterial DNAs. A pan-orthopox electrochemiluminescence (ECL) assay was used to further confirm the presence of Orthopoxvirus infection of the rodents. Seven of 12 (58%) animals (seven of 52 (15%) of all animals) tested positive in both monkeypox-specific PCR assays and two additional pan-orthopox PCR assays (in at least one tissue). The ECL results showed varying degrees of agreement with PCR. One hamster and three gerbils were positive by both PCR and ECL for all tissues tested. In addition, we attempted to verify the presence of monkeypox virus by culture on multiple cell lines, by immunohistology, and by electron microscopy, with negative results. Sequencing the PCR products from the samples indicated 100% identity with monkeypox virus strain Zaire-96-I-16 (a human isolate from the Congo). These real-time PCR and ECL assays represent a significant addition to the battery of tests for the detection of various orthopoxviruses. In light of the recent monkeypox virus transmissions, early detection of the virus is crucial for both natural outbreaks and potential acts of bioterrorism.

Animals↗
PubMedDate not supplied

Genetic heterogeneity in the VP2 gene of infectious bursal disease viruses detected in commercially reared chickens.

The genetic heterogeneity of infectious bursal disease virus (IBDV) vaccine strains was compared with IBDV detected in bursa tissue of commercially reared chickens. The vaccine strains tested represented classic viruses from the United States, South Africa, England, and France plus variant viruses from the United States. Bursa tissue samples used for the detection of IBDV from commercially reared chickens were from the United States. Genetic heterogeneity was examined using restriction fragment length polymorphisms (RFLP) of a 743-bp fragment of the VP2 gene that was amplified using reverse transcriptase/polymerase chain reaction (RT/PCR). The RT/PCR products were digested using restriction enzymes BstNI and MboI. On the basis of RFLP profiles, viruses were placed into molecular groups. Thirty-eight vaccine and laboratory strains of IBDV were placed into five molecular groups. Groups 1 and 2 contained variant viruses, groups 3 and 4 contained classic viruses, and group 5 contained Lukert/Edgar strain classic viruses. In contrast to these five molecular groups, 22 molecular groups were observed for 70 IBDV-positive samples from U.S. chicken flocks. Twenty-two of the 70 IBDV-positive samples were placed into molecular groups 1 (2 samples), 2 (18 samples), and 5 (2 samples). Nineteen new RFLP profiles that did not match the five molecular groups observed for vaccine strains were detected. No viruses were observed to have RFLP profiles like molecular groups 3 and 4 observed in the vaccine strains. The genetic heterogeneity was greater among IBDV strains circulating in commercially reared poultry compared with vaccine strains of the virus.

Animals↗
PubMed1978

Preadsorption of boar semen with kaolin: increased efficiency of foot-and-mouth disease virus detection.

The boar semen-associated cytotoxic factor(s), but not the antiviral activity, were removed by adsorption with kaolin. Although foot-and-mouth disease virus was efficiently removed from medium by kaolin or kieselguhr, the virus was not removed from semen-virus mixtures. Because the cytotoxicity induced by boar semen apparently altered the ability of tissue culture cells to support virus replication, preadsorption with kaolin increased the probability of detecting this virus in semen samples.

Adsorption↗