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At least 127 records · Page 7Linked to original sources
PubMed1993

Herpes simplex virus detection from genital lesions: a comparative study using antigen detection (HerpChek) and culture.

The sensitivity of a rapid enzyme immunoassay test (HerpChek Direct Herpes Simplex Virus Antigen Test [DuPont Medical Products, Wilmington, Del.]) for the detection of herpes simplex virus (HSV) antigens in patient specimens was compared with HSV culture. HerpChek positivity for HSV occurred with 179 (65%) of 275 lesion specimens, whereas culture for HSV was positive for 145 (53%) lesions (P = 0.002). HerpChek was twice as sensitive as culture for the detection of HSV in late-stage lesions and was equivalent to culture for the detection of HSV in early lesions. We conclude that HerpChek provides greater sensitivity than culture for HSV detection in late-stage genital lesions.

Adolescent↗
PubMed2002

[A retrospective study of clinical manifestation and human papilloma virus detection of oral condyloma acuminate in children].

OBJECTIVE: The aim of this study is to study genotype, transmission, clinical and pathological characteristics, and prognosis of oral condyloma acuminate (CA) in children. METHODS: The authors retrospected the clinical characteristics and slices of HE staining of six cases which have been diagnosed as oral CA and, performed inmunohistochemistry (IHC) and in situ hybridization (ISH) analysis to detect the DNA of human papilloma virus in 5 cases. RESULTS: Oral CA often happened in the hard or soft plates of children of two-year-old. Most of them came from the families had been infected by human papilloma virus (HPV). Histological examination demonstrated that koilocytes were common in the upper spinous and corneal layers. HPV was detected in all cases. HPV16/18-E6 antigen was positive in 4 of 5 cases examined. The result of ISH only show one case was HPV6- and HPV11-positive, and the other case was HPV-positive, but HPV could not be detected when recurring. CONCLUSION: The pathogen leading to oral condyloma acuminate (CA) and the transmission way of children may be different from that of adults.

Child, Preschool↗
PubMed1985

Factors influencing the sensitivity of herpes simplex virus detection in clinical specimens in a simultaneous enzyme-linked immunosorbent assay using monoclonal antibodies.

A rapid simultaneous enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies was investigated for herpes simplex virus (HSV) detection. All HSV isolated (n = 127) were detected, whereas no response was obtained with HSV negative preparations. Equivalent results were obtained from 275 of 277 clinical specimens in the monoclonal ELISA and in an ELISA using polyclonal antibodies, confirming that appropriately selected monoclonal antibodies may be as efficacious as polyclonal antibodies in antibody-based assays. In clinical specimens, the rate of HSV detection (sensitivity) relative to tissue culture isolation was low for both assays, and the major factor responsible for this was the low concentration of virus present in some specimens. The sensitivity of ELISA obtained in routine use varied with different panels of unselected specimens and was related to the speed of development of the cytopathic effect. These results emphasise the need for caution in assigning a definitive sensitivity level to ELISA tests evaluated on different panels of specimens.

Antibodies↗
PubMed1985

Persistence of serum IgA antibodies to herpes simplex, varicella-zoster, cytomegalovirus, and rubella virus detected by enzyme-linked immunosorbent assays.

Enzyme-linked immunosorbent assays were used to detect IgG and IgA antibodies to herpes simplex virus (HSV), varicella-zoster virus (VZV), cytomegalovirus (CMV), and rubella virus in sera from 68 adult female gynaecological patients. Of the patients who had virus-specific IgG antibodies, the proportion who also had virus-specific IgA was 98% for HSV, 75% for VZV, 73% for rubella virus, and 42% for CMV. IgA antibodies to all four viruses were only found when specific IgG antibodies were also detected in the serum. These results suggest that virus-specific IgA may persist for several years; possible explanations for this are discussed.

Adult↗
PubMed1988

Comparison of primary rabbit kidney and MRC-5 cells and two stain procedures for herpes simplex virus detection by a shell vial centrifugation method.

By using a conventional tissue culture method as a standard, four shell vial centrifugation culture (SVC) formats were compared for herpes simplex virus (HSV) detection in 300 clinical samples. Both MRC-5 and primary rabbit kidney (PRK) cells were used in the conventional and SVC systems. In addition, both a direct monoclonal fluorescent antibody to HSV (MAb-FA; Syva Corporation, Palo Alto, Calif.) and an indirect HSV polyclonal antibody-horseradish peroxidase stain (poly-HRP; Difco Laboratories, Detroit, Mich.) were used to stain shell vials of both cell types. Conventional tubes were incubated for up to 7 days with daily examination for cytopathic effect, which was confirmed as HSV by staining with an MAb-FA. Shell vials were inoculated, centrifuged, incubated for 16 to 24 h, and stained directly with MAb-FA or indirectly with a poly-HRP stain. Of the 300 specimens examined, 82 (27%) were HSV positive by conventional tissue culture. PRK cells detected 81 (99%) positive specimens, compared with 74 (90%) specimens detected with MRC-5 cells. Of the 82 positive specimens by conventional culture, the SVC formats detected 68 by MRC-5 and MAb-FA, 74 by MRC-5 and poly-HRP, 64 by PRK and MAb-FA, and 77 by PRK and poly-HRP. Therefore, PRK stained by an indirect method with poly-HRP was the most sensitive of the SVC formats tested, detecting 94% of the positive specimens.

Animals↗
PubMed1999

Two-centre study comparing DNA preparation and PCR amplification protocols for herpes simplex virus detection in cerebrospinal fluids of patients with suspected herpes simplex encephalitis.

In a two-centre study, the routine DNA preparation and PCR amplification protocols were compared for herpes simplex virus (HSV) detection in cerebrospinal fluids (CSFs) of 43 patients with suspected herpes simplex encephalitis (HSE). The combined clinical, radiological and laboratory results indicated HSE in 6/43 (14%) patients. Discrepant PCR results between the two centres were obtained in 8 (18%) cases consisting of 5 false-positive and 3 false-negative results. Seven out of 8 (88%) discrepant results were associated with the method of CSF preparation using protease K digestion followed by heat inactivation. In contrast, CSF digestion with proteinase K followed by DNA purification on silica spin columns was better yielding discrepant PCR results in only 1 of 78 analyses (1.3%). The results point to the need for standardization and inter-laboratory quality control for routine clinical work.

DNA, Viral↗
PubMed1994

Dengue virus. Detection and pathogeny.

Sequences front a cDNA of dengue virus type 4 were cloned into transcription vectors. These sequences included the E, NS1, NS2A, NS2B, NS3 genes. RNA transcripts produced in vitro from these plasmids were used in hybridization assays to detect dengue viral sequences. With these RNA-probes we have been able to detect molecules of serotype-specific dengue 4 viral RNA. Moreover, the riboprobes detected viral sequences of other serotypes in the following order of sensitivity 4 > 2 > 3 > 1, and might be useful to differentiate serotypes.

Animals↗
PubMed1993

High incidence of Epstein-Barr virus detection in Hodgkin's disease and absence of detection in anaplastic large-cell lymphoma in children.

By in situ hybridization with EBER oligonucleotides and immunohistochemistry with anti-latent membrane protein 1 (LMP1) antibody, we compared the detection rate of Epstein-Barr virus (EBV) in Hodgkin's disease and anaplastic large-cell lymphomas in children. Among the 13 cases of Hodgkin's disease tested, 7 (54%) were found to be EBV associated (EBER transcripts +, LMP1 +). None of the 11 cases of ALC lymphomas was found to contain EBV genomes or gene products. This may indicate that EBV is not a pathogenic agent in anaplastic large-cell lymphomas in children in comparison to Hodgkin's disease.

Adolescent↗
PubMedDate not supplied

[Hepatitis E virus: detection of antibodies in blood donors in Lebanon].

One hundred Lebanese volunteers blood donors were tested for the qualitative detection of antibodies against hepatitis E virus (HEV) by an immunoenzymatic method. The results showed 4% of positivity. This figure allows us to confirm the presence of infection due to HEV in Lebanon. More studies on a larger panel are necessary to confirm this result. This study leads us to suspect an HEV infection in front of an acute non A, non B, non C hepatitis even if the patients have not lived in an endemic area.

Adult↗
PubMed2003

Rapid preparation of DNA microarray using PCR for hepatitis B and D virus detection.

OBJECTIVE: To prepare DNA microarray for detecting both hepatitis B and D virus as (HBV and HDV). METHODS: With the assistance of Oligo6.4 software, specific PCR primers targeting the conserved region of HBV and HDV were designed. The PCR products were purified and cloned into the pMD18-T vectors, followed by rapid identification. The recombinant plasmids were then extracted from positive clones and the target gene fragments underwent sequence analysis. RESULTS: The gene fragments of HBV and HDV were obtained by PCR, which were confirmed by sequence analysis to be specific gene fragments of HBV and HDV. CONCLUSION: Using PCR amplification products to prepare the DNA microarray is quick, simple and effective.

Hepatitis B virus↗
PubMed1988

Effect of high-speed rolling on herpes simplex virus detection and replication.

We examined the effect of high-speed rolling on herpes simplex virus replication. Inoculated cultures were rolled at 2, 96, or 383 rpm, while stationary cultures served as controls. At 24 h, inoculated cultures rolled at 96 rpm had a 6.8-fold increase in foci when compared with stationary cultures (P less than 0.01) and a 2.8-fold increase over cultures rolled at 2 rpm (P less than 0.05). Cultures rolled at 2 rpm had a 2.4-fold increase in foci over stationary cultures (P less than 0.05). Viral yield results correlated with focus results. Significantly more virus was present in cultures rolled at 96 rpm (7.3-fold) than in stationary cultures. Cultures rolled at 2 rpm produced 2.9-fold more virus than stationary cultures (P less than 0.05). Of 37 cultures rolled at 96 rpm, 33 (89%) were cytopathic effect positive at 96 h, while 18 of 40 (45%) were positive at 2 rpm and only 2 of 37 (5%) were positive for stationary cultures (P less than 0.01). Cultures rolled at 96 rpm produced maximum viral yields 2 days sooner than stationary cultures. Rolling of inoculated cultures should be used in the clinical laboratory to aid in the rapid detection of herpes simplex virus.

Animals↗
PubMedDate not supplied

Comparison of dengue infection in human mononuclear leukocytes with mosquito C6/36 and mammalian Vero cells using flow cytometry to detect virus antigen.

Fluorescent activated cell sorter (FACS) analysis is useful for the detection of cellular surface antigens and intracellular proteins. We used this methodology in order to detect and quantify dengue antigens in highly susceptible cells such as clone C6/36 (Aedes albopictus) and Vero cells (green monkey kidney). Additionally, we analyzed the infection in vitro of human peripheral blood mononuclear leukocytes (PBML). FACS analysis turned out to be a reliable technique to quantify virus growth in traditional cell cultures of C6/36 as well as Vero cells. High rates of infection were achieved with a good statistical correlation between the virus amount used in infection and the percentage of dengue antigen containing cells detected in infected cultures. We also showed that human monocytes (CD14+) are preferred target cells for in vitro dengue infection among PBML. Monocytes were much less susceptible to virus infection than cell lines but they displayed dengue antigens detected by FACS five days after infection. In contrast, lymphocytes showed no differences in their profile for dengue specific immunofluorescence. Without an animal model to reproduce dengue disease, alternative assays have been sought to correlate viral virulence with clinical manifestations and disease severity. Study of in vitro interaction of virus and host cells may highlight this relationship.

Animals↗
PubMed2002

Flow cytometry assay for intracellular rabies virus detection.

Following previous studies reporting microbiological diagnosis by flow cytometry, the possibility of using this method was examined to monitor infection of susceptible cell lines by a fixed rabies virus strain (Pasteur Virus strain-PV) or a wild rabies virus strain (WRS). Suspensions of BHK-21 and C6 cells were infected with viruses and a time course of virus infection was established. Sequentially, at several time points, infected and control uninfected cells were fixed, permeabilized, and stained with a rabies virus-specific antibody conjugate. This was achieved by resuspending cells in a solution containing p-formaldehyde in FACS lysis fluid, which allowed the detection of intracellular virus with flourescein-coupled antibodies by flow cytometry. A Becton Dickinson FACSCalibur flow cytometer was used to analyze the percentage of cells infected and the kinetics of the infection process was determined. As early as 12 h after inoculation with both rabies virus strains, significant levels (P<0.01) of infection (from 4.7 to 7.1%) were detected by flow cytometry. The maximum level of infection was obtained at 48 h in C6 cells (88%) with both viruses. The advantages of this method for examination of intracellular virus infection are discussed.

Animals↗
PubMedDate not supplied

Sensitive non-radioactive nucleic acid hybridization assay for plum pox virus detection.

A new non-radioactive sandwich hybridization assay was designed to simplify the analysis of a large number of plant samples. Plant material was homogenized in 0.5% SDS and added directly to the hybridization reaction, in which a pair of identifying probes were used. One of the probes was biotinylated capture RNA specific for plum pox virus (PPV) strain SK-68; the other RNA probe was synthesized from a plasmid bearing the adjacent sequence of this strain and was labelled with digoxigenin (DIG). Both purified viral RNA and crude extracts from PPV-infected plants were used as target for sandwich hybridization. The hybridization reaction was carried out in a streptavidin-coated ELISA plate. After extensive washing, the viral RNA was detected by conventional colour reaction using anti-DIG/alkaline phosphatase conjugate. In comparative experiments, we have shown that this non-radioactive detection system is more sensitive than conventional ELISA techniques and we were able to detect virus-specific RNA in more than 50% of the ELISA-negative samples.

Enzyme-Linked Immunosorbent Assay↗
PubMed2005

Phylogenetic characterization of canine distemper viruses detected in naturally infected dogs in North America.

In 2004, six puppies and one adult dog from a total of four premises were subjected to necropsy evaluation. For five of the seven dogs, disease caused by canine distemper virus (CDV) infection was suspected based on clinical signs. In all of the dogs, a diagnosis of CDV infection was established by the presence of compatible gross and histologic lesions, immunohistochemical labeling for CDV antigen, and detection of CDV RNA by reverse transcription-PCR. To further characterize the CDV strains detected in the four cases, complete gene sequences were determined for the hemagglutinin (H) and fusion (F) protein genes, while partial gene sequencing was performed for the phosphoprotein gene. A total of 4,508 bases were sequenced for the CDV strains detected from each of the four cases. Two cases were found to have identical sequences except for 2 bases in the intergenic region of the F and H genes. Phylogenetic analysis strongly suggested an evolutionary relationship between sequences detected in these two cases and those of phocine distemper virus 2 and two other strains of CDV not previously detected in the continental United States. Clear phylogenetic relationships were not established for viruses detected in the two additional cases; however, one strain showed similarity to CDV strains detected in a panda from China. Importantly, the three CDV strains detected were demonstrated to be genetically distinct from known vaccine strains and strains previously reported in the continental United States.

Amino Acid Sequence↗