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At least 109 records · Page 6Linked to original sources
PubMed1998

Epstein-Barr virus detection in kidney biopsy specimens correlates with glomerular mesangial injury.

To determine the relationship between the detection of Epstein-Barr virus (EBV)-specific DNA and glomerular injury, 33 renal needle-biopsy specimens that had been formalin-fixed and paraffin-embedded were analyzed using polymerase chain reaction (PCR) with subsequent nonradioactive Southern blot technique. Light microscopic examination and immunofluorescence were also performed. In 30 of 33 renal biopsy specimens, the beta globin gene could be successfully amplified as integrity controls. These 30 patients consisted of 12 patients with immunoglobulin A nephropathy (IgAN), 10 patients with minor glomerular abnormalities, 6 patients with membranous nephropathy, and 2 patients with focal/segmental lesions. EBV was detected in 7 of 12 patients with IgAN (58%), 3 of 6 patients with membranous nephropathy (50%), 0 of 10 patients with minor glomerular abnormalities (0%), and 2 of 2 patients with focal/segmental lesions. EBV detection was not disease specific. The EBV detection ratio of the group with glomerular mesangial lesions (64%; 9 of 14 patients) was significantly greater than those without (19%; 3 of 16 patients; P < 0.012, chi-square test). The EBV detection ratio of the group with glomerular lesions (60%; 12 of 20 patients) was significantly greater than those without (0%; 0 of 10 patients; P < 0.0016, Fisher's exact test), and the EBV detection ratio of the group with fibrinogen deposits observed in immunofluorescence (73%; 11 of 15 patients) was significantly greater than those without (7%; 1 of 15 patients; P < 0.0002, chi-square test). The EBV detection ratio of the group with immunoglobulin deposits (57%; 12 of 21 patients) was also significantly greater than those without (0%; 0 of 9 patients; P < 0.0040, Fisher's exact test). These data suggest that EBV can damage the glomerular mesangium beyond disease units and be mediated by immunoglobulin in patients with various chronic glomerulonephritides.

Biopsy, Needle↗
PubMed1989

Development of an enzyme-linked immunosorbent assay for equine infectious anemia virus detection using recombinant Pr55gag.

To provide more sensitive and convenient methods for the detection of equine infectious anemia virus (EIAV), we developed an enzyme-linked immunosorbent assay (ELISA) employing the EIAV gag precursor (Pr55gag) produced by using recombinant DNA techniques. The antigenic reactivity of the recombinant EIAV Pr55gag was found to be equivalent to that of the virion p24gag and elicited high-titered antiserum in rabbits. When a large number of horse sera were analyzed for the presence of antibodies to EIAV by this ELISA, a radioimmunoassay for EIAV p15gag, or the standard agar gel immunodiffusion test, there was 98.7% concordance among the assays. By using the ELISA it was possible to specifically detect antibodies earlier after experimental infection of horses with EIAV than with the other two tests. A competition ELISA developed in order to detect EIAV gag antigens was found to be approximately 15 times more sensitive than the radioimmunoassay for EIAV p15gag. Antigens of other animal lentiviruses as well as those of the prototype oncovirus failed to compete in this assay.

Animals↗
PubMed1975

Morphological methods of Kilham virus detection.

Morphological investigations on organs from Kilham virus (KV)-infected Syrian hamsters revealed acute glomerulonephritis with a clear-cut haemorrhagic syndrome. No pathological changes were found in internal organs from KV-infected rats.

Animals↗
PubMed2004

Wild rabies virus detection by plaque assay from naturally infected brains in different species.

A simple, sensitive and specific plaque assay protocol for the detection of wild type rabies virus in different species is described using confluent monolayers of chicken embryo cells in 6-well plates. Plaques are produced after application of either agarose or Sephadex G-100 overlay onto cell monolayers and incubation for 96 h after virus infection at 37 degrees C. The parameters affecting plaque appearance include cell seeding concentration, overlay composition and time of incubation after infection. Optimal conditions are seeding at a concentration of 4 x 10(6) cell/cm(3), incubation at 37 degrees C in 5% CO(2) atmosphere during 96 h, using either 1% agarose or 2% Sephadex G-100 overlays. The described plaque assay would be a new valuable tool in conducting various quantitative investigations, since the chicken embryo cells are susceptible to rabies virus infection from all species studied.

Animals↗
PubMed1999

A fast, sensitive and specific method for rice dwarf virus detection by northern blot hybridization.

Based on stability of double-stranded (ds) RNA, a new, fast, sensitive, and specific method for detection of genomic rice dwarf virus (RDV) dsRNA by molecular hybridization was developed. In contrary to the commonly used, standard Northern blot analysis, dsRNA is denatured in the immobilized state on the blot. Therefore, risk of degradation of single-stranded (ss) RNA by ribonuclease (RNase) during sample preparation, electrophoresis and blotting is eliminated. This method overcomes disadvantage of incomplete denaturation of dsRNA in Northern blot analysis. In conclusion, the newly developed method is reliable, sensitive, very specific, and gives a low background. The entire procedure is also less time consuming; it can be completed within 2-3 days. The new method may be regarded as a modification of the standard Northern blot analysis.

Blotting, Northern↗
PubMed2006

JC virus detection in bodily fluids: clues to transmission.

JC virus in saliva, oropharyngeal fluid, blood, and urine samples obtained from 58 human immunodeficiency virus-infected persons and 58 matched controls was investigated by performing quantitative polymerase chain reaction. JC virus was rarely present in oropharyngeal fluid and blood samples, even in those obtained from immunosuppressed individuals, but it was commonly detected in urine samples from both groups, suggesting that urine contributes to transmission.

Adult↗
PubMed2002

Partial nucleotide sequencing of hepatitis E viruses detected in sera of patients with hepatitis E from 14 cities in China.

OBJECTIVE: To investigate the genotypes of hepatitis E viruses (HEV) detected in sera of patients from different regions of China. METHODS: The partial genome (nt6461-6860, nt5994-6294) of open reading frame 2 (ORF2) of 45 HEV strains detected from 14 cities of China was amplified and sequenced using polymerase chain reaction (PCR) and direct sequencing. RESULTS: Forty-one of 45 strains (91%) share the same genotype with HEV Burma strain (B), with nucleotide identities higher than 98% with the representative HEV Chinese strain. Only 4 HEV strains are significantly divergent from the 3 prototype strains of HEV, with nucleotide identities of 77%-80% with HEV Burmese/Chinese strain, 74%-76% with Mexican strain and 74%-77% with the newly discovered HEV US/swine strain, respectively. Phylogenetic analysis suggests that these 4 strains may represent 2 different subtypes that belong to a novel genotype of HEV, which is significantly divergent from the prototype Mexico, Burmese and US/swine strains. CONCLUSION: Among patients with hepatitis E in China, most are infected by the Chinese prototype HEV, and only a small part by the new genotype HEV.

Base Sequence↗
PubMed2004

Smallpox and pan-orthopox virus detection by real-time 3'-minor groove binder TaqMan assays on the roche LightCycler and the Cepheid smart Cycler platforms.

We designed, optimized, and extensively tested several sensitive and specific real-time PCR assays for rapid detection of both smallpox and pan-orthopox virus DNAs. The assays are based on TaqMan 3'-minor groove binder chemistry and were performed on both the rapid-cycling Roche LightCycler and the Cepheid Smart Cycler platforms. The hemagglutinin (HA) J7R, B9R, and B10R genes were used as targets for the variola virus-specific assays, and the HA and DNA polymerase-E9L genes were used as targets for the pan-orthopox virus assays. The five orthopox virus assays were tested against a panel of orthopox virus DNAs (both genomic and cloned) at the U.S. Army Medical Research Institute of Infectious Diseases (USAMRIID). The results indicated that each assay was capable of detecting both the appropriate cloned gene and genomic DNA. The assays showed no cross-reactivity to the 78 DNAs in the USAMRIID bacterial cross-reactivity panel. The limit of detection (LOD) of each assay was determined to be between 12 and 25 copies of target DNA. The assays were also run against a blind panel of DNAs at the Centers for Disease Control and Prevention (CDC) on both the LightCycler and the Smart Cycler. The panel consisted of eight different variola virus isolates, five non-variola virus orthopox virus isolates, two varicella-zoster virus isolates, and one herpes simplex virus isolate. Each sample was tested in triplicate at 2.5 ng, 25 pg, 250 fg, and 2.5 fg, which represent 1.24 x 10(7), 1.24 x 10(5), 1.24 x 10(3), and 1.24 x 10(1) genome equivalents, respectively. The results indicated that each of the five assays was 100% specific (no false positives) when tested against both the USAMRIID panels and the CDC blind panel. With the CDC blind panel, the LightCycler was capable of detecting 96.2% of the orthopox virus DNAs and 93.8% of the variola virus DNAs. The Smart Cycler was capable of detecting 92.3% of the orthopox virus DNAs and between 75 and 93.8% of the variola virus DNAs. However, all five assays had nearly 100% sensitivity on both machines with samples above the LOD (>12 gene copies). These real-time PCR assays represent a battery of tests to screen for and confirm the presence of variola virus DNA. The early detection of a smallpox outbreak is crucial whether the incident is an act of bioterrorism or an accidental occurrence.

Base Sequence↗
PubMed1998

Epstein-Barr virus detection in nasopharyngeal tissues of patients with suspected nasopharyngeal carcinoma.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is strongly associated with Epstein-Barr virus (EBV). Detection of EBV in biopsy specimens may serve as a tumor marker. METHODS: To assess the sensitivity and specificity of the polymerase chain reaction (PCR) and in situ hybridization in the diagnosis of NPC, formalin fixed, paraffin embedded nasopharyngeal biopsies from patients in Taiwan suspected of having NPC were studied. RESULTS: In specimens from 107 patients with NPC, EBV was detected by PCR in 97 cases (90.7%) and by EBER in situ hybridization in 105 cases (98.1%). In specimens from 61 patients without neoplasia, EBV was detected by PCR in 7 cases (11.5%) and by EBER in situ hybridization in 0 cases. CONCLUSIONS: These results suggest that although EBV DNA may occasionally be detected in nonneoplastic nasopharyngeal tissues, cells expressing EBER are not. EBER in situ hybridization may therefore prove to be a useful adjunct in the diagnosis of NPC.

Biopsy↗
PubMed2002

[Evaluation of Norwalk virus detection kit by enzyme immunoassay].

We briefly examined detection kit using the EIA method for Norwalk virus, and compared the results of the tests using the EIA method with those using RT-PCR method. In reproducibility, an amount of variation was observed in data obtained from positive controls and in lower values. The sensitivity obtained from the EIA method was about 300 times lower than that obtained from the RT-PCR method. Results accordance ratio between EIA method and RT-PCR method was 70%. This results discrepancy was presumably caused by a difference of sensitivity and specificity between these two methods. In conclusion, this detection kit using the EIA method is easily manually operated so that this kit can be considered as an effective and simple detection tool for Norwalk virus.

Immunoenzyme Techniques↗
PubMed1992

Lack of correlation between virus detection and serologic tests for diagnosis of active cytomegalovirus infection in patients with AIDS.

The aim of this study was to evaluate the usefulness of serologic analysis for the diagnosis of active cytomegalovirus infection in patients with AIDS. Active cytomegalovirus infection was diagnosed by virus isolation from urine and saliva and detection of antigenemia. Serologic analysis was done by several conventional and innovative procedures. The results indicate no correlation between any of the most popular serologic procedures and virus detection by culture in urine or saliva or antigenemia.

Acquired Immunodeficiency Syndrome↗
PubMed1998

Hepatitis A virus detection in wastewater by PCR and hybridization.

Hepatitis A virus is a member of the Picornaviridae family and is a principal agent of acute hepatitis worldwide, causing from mild to severe illness. Although the incidence of hepatitis A is in decline, the risk of this disease is still high in the Mediterranean area. Detection of hepatitis A in the environment is difficult because this virus needs a prolonged incubation in cell culture, therefore we used an antigen capture PCR (AC-PCR) followed by a hybridization on membrane to identify HAV in wastewater samples. The raw sewage, concentrated by ultrafiltration, showed 8 positive samples out of 10 (80%), while after the oxidation step of the sewage, 2 out of 10 (20%) and 3 out of 10 (30%) were found positive respectively after concentration by electronegative (HAWP Millipore) and electropositive (1MDS Cuno-Div.) membranes. In the final effluent the positivity was 1 out of 10 (10%) for the electronegative membranes and 3 out of 10 (30%) for the electropositive membranes. Our results indicate: i) the possibility of HAV to cross the wastewater treatment plant and contaminate water and food (such as mussels); ii) PCR-hybridization as a rapid method for HAV identification in the environment.

Hepatitis A↗
PubMed2002

[Newly developed Norwalk-like viruses detection ELISA by combination with monoclonal antibodies and hyperimmune rabbit sera].

Norwalk-like viruses(NLVs) antigen detection ELISA was newly developed. It is constructed with monoclonal antibodies(MAbs) as a capture antibodies and hyperimmune rabbit sera as a detector antibodies. The two MAbs reacted broadly and specifically with genogroup I and genogroup II NLVs, respectively. Total detection time is less than 3 hours and at least 90 stool samples could be tested at one time. The concordance rate with ELISA and RT-PCR was 69% and it could be raised to 71% if capsid primer sets were used. Epidemiological study revealed that among 664 normal persons one sample(0.2%) was confirmed positive by both ELISA and RT-PCR, suggested that NLV carrier may exist in the normal person. The ELISA kit has advantages such as rapid, simple and multi samples detection, and useful for the first screening test. Also it is big tool for epidemiological surveillance of NLVs invasions.

Animals↗
PubMed2003

Respiratory viruses detected in hospitalised paediatric patients with respiratory infections.

Over 200 strains of respiratory viruses cause a variety of human infections ranging from common cold to life-threatening pneumonia. Respiratory viruses implicated in this study are respiratory syncytial viruses (RSV), adenovirus, influenza viruses and parainfluenza viruses. The objective of this study is to determine the epidemiology of respiratory viruses in paediatric patients with lower respiratory tract infection. The methods used were direct antigen detection method, shell vial culture method and conventional tube culture method. The samples included in this study are paediatric patients seen in Universiti Kebangsaan Malaysia Hospital, Kuala Lumpur with suspected acute viral respiratory infection, presenting with acute laryngotracheobronchitis (croup), bronchiolitis and pneumonia. Nasopharyngeal aspirates were collected and processed almost immediately. A total of 222 specimens were received during February 1999 to January 2000 showing a dual peak pattern in the months of April and December. The mean age of the patients was 13 months. Pneumonia (77.9%) was the most common clinical diagnosis in children with lower respiratory tract infection. This was followed by bronchiolitis (19.4%) and croup (27%). Viral aetiologies were confirmed in 23.4% of the patients. The most common respiratory virus isolated or detected was RSV, followed by parainfluenza viruses, influenza viruses and adenovirus.

Bronchiolitis, Viral↗
PubMed2006

Characterisation of the discrepancy between PCR and virus isolation in relation to classical swine fever virus detection.

In order to confirm and characterise further the discrepancies observed between diagnostic RT-nPCR and virus isolation results for the detection of classical swine fever virus (CSFV), a test panel of three new RT-PCRs was designed, amplifying parts of the NS2, NS3 and NS5A regions. Screening of negative samples by virus isolation with the new panel not only confirmed the discrepancies previously observed but also indicated that these were not associated with a specific genomic region. However, none of the PCR-positive samples were positive on all the different PCRs and preferential amplification was not obtained even when a more sensitive real-time RT-PCR was used. Furthermore, the primer-dependent amplification, most likely caused by the presence of viral fragments, demonstrates the necessity of confirming a single positive PCR result, certainly in the presence of contradictory virus isolation results. The new PCR panel, in combination with sequencing, can be used as a tool to provide additional information on the nature of the viral RNA present in the sample.

Animals↗
PubMed2001

Development of a highly sensitive nested RT-PCR method for Beet necrotic yellow vein virus detection.

A diagnostic test incorporating reverse-transcription polymerase chain reaction (RT-PCR) and nested polymerase chain reaction (nPCR) was developed for the detection of Beet necrotic yellow vein virus (BNYVV). The RT-PCR used the primers designed by (Henry et al., J. Virol. Methods 54 (1995)15) but refinements were made to the protocol including simplification of the extraction method, the use of standard reagents and adoption of a one-step procedure. None of these changes impaired sensitivity or specificity. The RT-PCR could also be used to amplify immunocaptured virus but this was slightly less sensitive than amplification from purified RNA. In nPCR, a second round of amplification was performed using primers, which produce a specific 326 base-pair product. Both RT-PCR and nPCR detected a range of 21 isolates collected from Europe, America and Asia (including A, B and P pathotypes) isolated from either sugar beet or Chenopodium quinoa. Neither assay produced PCR products using total RNA extracted from the roots of healthy sugar beet or beet infected with Beet soil-borne virus. However, the sensitivity of the nPCR was 1000 times greater than the standard RT-PCR. The reliability of the standard RT-PCR and nPCR was demonstrated using a range of cultivars collected from an infected field site. The use of the nPCR assay is recommended for applications where its improved sensitivity over standard RT-PCR is necessary, for example in the early detection of infection from bait-test soils and for quarantine and breeding purposes.

Chenopodiaceae↗
PubMed2004

An ELISA optimized for porcine epidemic diarrhoea virus detection in faeces.

Monoclonal antibodies to porcine epidemic diarrhoea virus (PEDV) membrane protein M were prepared and used for the comparative assessment of three blocking ELISA variants to detect PEDV. The competitive blocking ELISA (CB-ELISA) format showed the highest sensitivity, allowing detection of 10(2.5) plaque-forming units of PEDV/ml in culture medium. Its specificity was verified by inclusion of control samples containing transmissible gastroenteritis virus (TGEV) and rotavirus A in each analysis. Eighty porcine field samples of faeces obtained from 38 herds affected with diarrhoea were examined, and PEDV was found in 15 (19%) samples from 6 (16%) herds. The suitability of the CB-ELISA for the screening herds in epizootiologic situations is discussed.

Animals↗
PubMed1990

[Ultrastructural papilloma virus detection in precancerous and early stages of cervix cancer].

100 women with pathologic findings in the uterine cervix are the basis of this study. To assess papilloma virus articles methods of electron microscopy have been used in specimens of 59 women if koilocytes were present. Inside the nucleus of koilocytes in 51 cases virus particles could be detected, in 31 cases isolated and in 20 cases agglomerated between the chromatin. In all 10 condylomas without nuclear atypia we could assess virus particles. With increasing atypia the detection of papilloma virus decreases.

Biopsy↗