Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:2.7.7.49 (reverse transcriptase)
31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The study was divided into two experiments. In the first experiment, the efficacy of in ovo intermediate vaccine against infectious bursal disease virus (IBDV) was determined by challenge at 21 days of age with virulent IBDV in specific-pathogen-free (SPF) and commercial chickens. This vaccine was able to induce active immunity and to protect SPF chickens to challenge; protection was not complete in commercial chickens, as testified by bursal lesions, bursal index after challenge, and vaccine immunoresponse. In order to detect field and vaccinal viruses, immunoperoxidase staining, enzyme-linked immunosorbent assay, capture, and reverse transcriptase-polymerase chain reaction (RT-PCR) were tested; the RT-PCR was more effective at detecting both kind of viruses. In the second experiment, the immunosuppressive effect of in ovo vaccination was determined by evaluating the immunoresponse against Newcastle disease virus (NDV) vaccination effected at 10 days in both SPF and commercial chickens vaccinated in ovo. The in ovo vaccine causes a reduction of NDV immunoresponse, as testified by lowest geometric mean titer in group I (SPF chickens vaccinated against IBDV in ovo and against NDV at 11 days). In commercial chickens, immunoresponse to NDV vaccination was not influenced by in ovo vaccination.
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PMID:Efficacy and safety of an infectious bursal disease virus intermediate vaccine in ovo. 1178 75

We genetically analyzed field isolates of the Newcastle disease (ND) virus isolated in Japan from 1930 to 2001. The coding region of the fusion protein was amplified by reverse transcriptase PCR and directly sequenced. Phylogenetic analysis revealed the presence of viruses belonging to six of the eight known genotypes. It can be concluded from this study that ND outbreaks in Japan have been of multiple etiologies. [All sequences used in this study were sent to DDBJ and assigned accession numbers AB 070382 to AB 074042.]
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PMID:Phylogenetic analysis of Newcastle disease virus genotypes isolated in Japan. 1235 91

A method is described for enabling safe transit of denatured virus samples for polymerase chain reaction (PCR) identification without the risk of unwanted viable viruses. Cotton swabs dipped in avian infectious bronchitis virus (IBV) or avian pneumovirus (APV) were allowed to dry. Newcastle disease virus and avian influenza viruses were used as controls. Autoclaving and microwave treatment for as little as 20 sec destroyed the infectivity of all four viruses. However, both IBV and APV could be detected by reverse transcriptase (RT)-PCR after autoclaving and as long as 5 min microwave treatment (Newcastle disease virus and avian influenza viruses were not tested). Double microwave treatment of IBV and APV with an interval of 2 to 7 days between was tested. After the second treatment, RT-PCR products were readily detected in all samples. Swabs from the tracheas and cloacas of chicks infected with IBV shown to contain infectious virus were microwaved. Swabs from both sources were positive by RT-PCR. Microwave treatment appears to be a satisfactory method of inactivating virus while preserving nucleic acid for PCR identification.
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PMID:Microwave or autoclave treatments destroy the infectivity of infectious bronchitis virus and avian pneumovirus but allow detection by reverse transcriptase-polymerase chain reaction. 1522 57

The complete nucleotide sequence of the gene encoding the matrix protein (M) of the avian paramyxovirus, serotype 3b (APMV-3b), has been determined by means of the direct sequencing of viral RNA using reverse transcriptase reaction. The adjacent portions of the neighboring phosphoprotein (P) and fusion (F) protein genes were also sequenced that permitted to determine the consensus sequence of the viral genome, the poly(A) tract, downstream and upstream non-coding portions of the P and F genes, respectively, as well as the corresponding intergenic regions. The gene is 1478 nucleotides long with a protein-coding sequence of 1194 nucleotides. The deduced protein consists of 398 amino acids with a calculated MW 44,465. According to the multalignment and phylogenetic analyses, the APMV-3b M protein has shown the closest relatedness towards Newcastle disease virus (NDV) which has recently been suggested to be excluded from the Rubulavirus genus and assigned (together with APMV-6) to a new Avulavirus genus within the subfamily Paramyxovirinae of the Paramyxoviridae family. On the basis of the M protein genetic multalignment, phylogenetic relationships, bipartite nuclear localization signal identification in combination with the cysteine residues distribution, and by the degree of intrageneric heterogeneity, the APMV-3b is proposed to be another member (together with NDV and APMV-6) of the new genus.
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PMID:Nucleotide sequence of the matrix protein gene of avian paramyxovirus, serotype 3b: evidence on another member of the suggested new genus of the subfamily Paramyxovirinae. 1556 52

Vaccination for Newcastle disease (ND) is routinely practised in countries where virulent strains of the Newcastle disease virus (NDV) are endemic and in countries where virulent strains do not exist but ill-timed infection by a low virulent field strain may have significant economic consequences for the producer. The types of vaccines and vaccination schedules used vary depending on the potential threat, virulence of the field challenge virus, type of production, and production schedules. A combination of live and inactivated ND vaccine, administered simultaneously, is shown to provide better protection against virulent NDV and has been successfully used in control programmes in areas of intense poultry production. A potential limiting factor in the use of live vaccines to control virulent ND is that live virus can interfere with surveillance and laboratory diagnosis. However, a new assay, the real-time reverse transcriptase-polymerase chain reaction (RRT-PCR), differentiates low virulent from virulent NDV, thus minimizing the disadvantage of live virus vaccines in the face of an outbreak and may facilitate the use of such vaccines to control outbreaks of virulent ND in the future.
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PMID:Control of Newcastle disease by vaccination. 1574 28

During the 2002--2003 Exotic Newcastle Disease (END) outbreak in Southern California, a high-throughput real-time reverse transcriptase-polymerase chain reaction (RRT-PCR) system was developed to respond to the large diagnostic and surveillance sample workload. A 96-well RNA extraction method, using magnetic bead technology, combined with a 96-well RRT-PCR assay, allowed 1 technician to process and test more than 400 samples per day. A 3-technician team could complete testing on approximately 1,900 samples per day. The diagnostic sensitivity of the high-throughput RRT-PCR assay was 0.9967 (95% CI 0.9937-0.9997) based on 926 virus isolation confirmed positive samples. Diagnostic specificity using an initial 434 virus isolation confirmed negative samples was 100%. A diagnostic specificity of 0.9999 (95% CI 0.9999, >0.9999) was subsequently calculated on the basis of 2 false-positive results among 65,343 surveillance samples collected after the final END-positive case was confirmed in May 2003. Assay performance over 500 replicates, including reproducibility of the combined extraction and RRT-PCR amplification steps yielded a standard deviation of 0.70 RRT-PCR cycle thresholds (Ct) and a standard deviation of 0.59 Ct for the RRT-PCR steps alone. The high-throughput RRT-PCR developed for END contributed significantly to the 2002--2003 END control effort, reducing the predicted timeline for eradication from 3 years to just 11 months, primarily because of the large number of samples that could be rapidly tested. The 96-well approach described for high-throughput END RRT-PCR could be adapted to other rapid, high-volume testing needs, as required for potential foreign animal disease responses or intensive surveillance efforts.
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PMID:High-throughput real-time RT-PCR assay to detect the exotic Newcastle Disease Virus during the California 2002--2003 outbreak. 1582 92

The 2002--2003 Exotic Newcastle Disease (END) outbreak in Southern California poultry provided an opportunity to evaluate environmental air sampling as an efficient and cost-effective means of sampling flocks for detection of a circulating virus. Exotic Newcastle Disease virus was detected by real-time reverse transcriptase PCR from air samples collected using a wetted-wall cyclone-style air sampler placed within 2 m of birds in 2 commercial flocks suspected of being naturally exposed to END virus during the outbreak. Exotic Newcastle Disease virus was detected after 2 hours of air sampling the poultry-house environments of the 2 naturally infected flocks.
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PMID:Environmental air sampling to detect exotic Newcastle disease virus in two California commercial poultry flocks. 1582 6

Proventriculitis was studied by experimentally reproducing the disease in broiler chickens. One-day-old infectious bursal disease virus (IBDV) and infectious bronchitis virus (IBV) antibody positive commercial broilers and 1-day-old antibody negative specific-pathogen-free (SPF) broilers were orally gavaged with proventricular homogenates produced from the proventriculi of broilers with proventriculitis. At 7 and 14 days, both commercial and SPF broilers had enlargement of the proventriculus with necrosis of the glandular epithelium and lymphocytic infiltrates in the proventricular glands. SPF broilers exposed to the proventricular homogenates developed infectious bursal disease, and IBDV was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). They also were positive by RT-PCR to IBV and developed nephritis. Commercial broilers developed mild nephritis but not bursal disease and were negative for IBDV and positive for IBV by RT-PCR. Both homogenate-inoculated commercial and SPF chickens were negative for reovirus and Newcastle disease virus by RT-PCR and variably positive for adenovirus by PCR. Bacteria were not identified in histologic sections, nor were they isolated from affected proventriculi. Indirect fluorescent antibody assay using convalescent sera detected intracytoplasmic staining in the proventricular glandular epithelial cells. Examination of thin sections of proventriculi using electron microscopy identified virus-like particles approximately 120 nm in diameter within the cytoplasm of these cells at 7 days after inoculation. Passage of proventricular homogenate filtrates in chicken embryos for virus isolation caused stunting, and allantoic fluid from these eggs was positive for IBV by RT-PCR.
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PMID:Reproduction of proventriculitis in commercial and specific-pathogen-free broiler chickens. 1625 87

The feasibility of using Flinders Technology Associates filter papers (FTA cards) to collect allantoic fluid and chicken tissue samples for Newcastle disease virus (NDV) molecular detection was evaluated. Trizol RNA extraction and one-step reverse transcriptase-polymerase chain reaction (RT-PCR) were used. FTA cards allowed NDV identification from allantoic fluid with a titre of 10(5.8) median embryo lethal doses/ml. The inactivated virus remained stable on the cards for 15 days. NDV was detected from FTA imprints of the trachea, lung, caecal tonsil and cloacal faeces of experimentally infected birds. RT-PCR detection from FTA cards was confirmed by homologous frozen-tissue RT-PCR and virus isolation. Direct nucleotide sequence of the amplified F gene allowed prediction of NDV virulence. No virus isolation was possible from the FTA inactivated samples, indicating viral inactivation upon contact. The FTA cards are suitable for collecting and transporting NDV-positive samples, providing a reliable source of RNA for molecular characterization and a hazard-free sample.
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PMID:Use of FTA filter paper for the molecular detection of Newcastle disease virus. 1659 99

The current available molecular method to detect infectious bursal disease virus (IBDV) is by reverse transcriptase-polymerase chain reaction (RT-PCR). However, the conventional PCR is time consuming, prone to error and less sensitive. In this study, the performances of Sybr Green I real-time PCR, enzyme-linked immunosorbent assay (ELISA) and conventional agarose detection methods in detecting specific IBDV PCR products were compared. We found the real-time PCR was at least 10 times more sensitive than ELISA detection method with a detection limit of 0.25pg. The latter was also at least 10 times more sensitive than agarose gel electrophoresis detection method. The developed assay detects both very virulent and vaccine strains of IBDV but not other RNA viruses such as Newcastle disease virus and infectious bronchitis virus. Hence, Sybr Green I-based real-time PCR is a highly sensitive assay for the detection of IBDV.
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PMID:Comparison of Sybr Green I, ELISA and conventional agarose gel-based PCR in the detection of infectious bursal disease virus. 1697 Nov 1


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