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)

Infectious bursal disease viruses (IBDVs) were examined for restriction fragment length polymorphisms in a fraction of the VP2 gene with the use of the reverse transcriptase/polymerase chain reaction-restriction fragment length polymorphism (RT/PCR-RFLP) assay. The restriction enzymes BstNI and Mbol were used to obtain RFLP results. A third enzyme, StyI, was tested, but its utility for differentiation of IBDV strains was limited. Thirteen vaccine viruses and five IBDV strains that were previously characterized were placed into five molecular groups. Two groups contained viruses described as being classic strains, and two groups contained viruses described as being variant strains. The fifth group contained both classic and variant strains. The RFLP observed for the serotype 2 IBDV strain OH was unlike any of the RFLPs observed in viruses in the five molecular groups. Seven IBDV strains from commercially reared chickens in the United States, Mexico, Puerto Rico, and Thailand were tested in the RT/PCR-RFLP assay to determine if they were similar to the commercial IBDV vaccine strains tested. These viruses were selected because they were associated with lesions in the bursa of chickens that should have been protected by maternal antibodies or active immunity. Each of the viruses tested contained a unique RFLP compared with the IBDV strains and vaccine viruses examined in this study and, thus, did not fit into any of the five molecular groups. These viruses also were distinguishable from each other.
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PMID:Restriction fragment length polymorphisms in the VP2 gene of infectious bursal disease viruses. 935 9

Infectious bursal disease viruses (IBDVs) were identified in bursa samples from chickens reared outside the United States. All samples were obtained from commercially reared chicken flocks experiencing signs typical of infectious bursal disease. The reverse transcriptase/polymerase chain reaction (RT/PCR) technique was used to detect the viruses and restriction fragment length polymorphism (RFLP) was used to compare a 743-bp region of the VP2 gene among the viruses detected. The 81 IBDVs detected were from 16 different countries and were assigned to molecular groups on the basis of their RFLP following digestion with the restriction enzymes BstNI and MboI. The presence of an SspI site in the 743-bp RT/PCR fragment was used to predict the very virulent (vv) phenotype. Almost half (49%) of the viruses detected had the same RFLP with the BstNI and MboI enzymes. These viruses were placed into a new molecular group, designated group 6, that was exemplified by the RS593 vaccine strain of IBDV, which also had this molecular RFLP pattern. Some viruses detected had RFLP patterns similar to molecular groups 2, 3, and 4, previously described in our laboratory. Sixteen RFLP patterns were observed that did not match RFLP results we previously obtained for vaccine IBDV strains. The SspI restriction site was found in 46% of the viruses detected, predicting that these viruses have the vvIBDV phenotype. The SspI site was not observed in viruses from Central and South America.
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PMID:Restriction fragment length polymorphisms in the VP2 gene of infectious bursal disease viruses from outside the United States. 1039 45

Infectious bursal disease virus (IBDV) strains have been identified and placed into molecular groups by a reverse transcriptase (RT)/polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) assay. The predicted amino acid sequences corresponding to the region of the genome examined by RFLP were determined and compared for 14 IBDV strains from different molecular groups and 11 IBDV strains that were identified in molecular group 6. Among the viruses within molecular group 6, 13 amino acid positions had mutations, and among the viruses in different molecular groups, 27 amino acid positions had mutations. In addition to having more mutations, viruses compared from different molecular groups also had mutations at key positions that were previously reported to be important for the formation of neutralizing epitopes. Three of these IBDV strains with unique RFLP patterns were used to challenge 1-wk-old broiler chickens with maternal immunity to IBDV. One of these viruses, T1, broke through this maternal immunity as evidenced by detection of the virus by RT/PCR-RFLP and production of an active virus neutralizing antibody response to classic and variant IBDV strains. Unique amino acid mutations in the T1 virus that may have contributed to its ability to break through this maternal immunity were observed at amino acids 318 and 322. The results indicate that RFLP profiles and nucleotide sequences can be used to predict the relative similarities and differences among IBDV strains, but determining the actual antigenic differences among viruses requires testing in vivo.
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PMID:Amino acid comparison of infectious bursal disease viruses placed in the same or different molecular groups by RT/PCR-RFLP. 1141 12

Infectious bursal disease virus is a contagious, immunosuppressive disease of young chickens that is controlled by vaccination. Cross-protection occurs between different strains of the virus as a result of shared neutralizing epitopes. However, interactions between two antigenically similar strains (a mild and a pathogenic) coinfecting the same host have not been investigated. Groups of specific-pathogen-free chickens were inoculated with a mild strain followed by a pathogenic strain at 0, 16, 24, or 48 hr postinoculation (PI) with a mild strain. Virus persistence and the predominant strain of the virus were determined by reverse transcriptase-polymerase chain reaction and restriction fragment length polymorphism analysis, respectively, in bursas at 2, 4, 8, 14, and 21 days PI with the pathogenic strain. Severity of infection was assessed by the bursa/body weight ratios and histopathologic lesion scores. The mild virus interfered with replication of the pathogenic virus. The greatest interference was observed when the pathogenic strain was inoculated 24 hr PI with the mild strain. The interference phenomenon observed might be due to competition for host receptor sites or production of cytokine(s). This interference phenomenon could have practical implications for vaccine usage and protection.
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PMID:Interference between mild and pathogenic strains of infectious bursal disease virus in chickens. 1583 20

Infectious bursal disease (IBD) associated with high mortality was first observed in Europe in the mid-1980s. The viruses identified in those outbreaks were described as being very virulent infectious bursal disease virus (vvIBDV) strains. These viruses have spread to nearly every continent but have not yet been identified in North America, Australia, and New Zealand. There is a real and immediate concern that the very virulent form of IBDV will continue to spread until it is present on every continent. Genomic RNA samples from IBDV strains suspected of being very virulent were submitted to our laboratory for molecular analysis. Nucleotide sequences of the VP2 gene hypervariable sequence region were determined for 18 of these viruses. A comparison with published vvIBDV sequences indicated that all but one sample (Thai 4) had nucleotide and predicted amino acid sequences consistent with vvIBDV strains. Published sequences and the nucleotide sequences of our 17 putative vvIBDV strains were used to identify unique nucleotides in the VP2 gene. Probe pairs for a real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assay were designed based on these unique sequences and then used to test the 17 genomic samples that were identified by nucleotide sequencing to be consistent with vvIBDV, plus the one Thai 4 sample that was not consistent with vvIBDV. Using melting temperature (Tm) analysis following real-time RT-PCR, two probe pairs (vv232 and vv256) successfully identified the 17 putative vvIBDV strains and distinguished them from the Thai 4 sample. An additional 26 genomic RNA samples submitted as suspect vvIBDV strains were then tested using the vv232 and vv256 probes. Based on the melting point analysis of these two probes, all 26 samples contained nucleotide sequences consistent with vvIBDV strains. The specificity of the vv232 and vv256 probe pairs was evaluated using 19 non-vvIBDV strains. In every case, the probes distinguished the 19 classic and variant (non-vvIBDV) strains from the putative vvIBDV strains. Diagnostic assays that can reliably identify vvIBDV strains are needed for surveillance programs designed to monitor the spread of these viruses.
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PMID:Molecular studies on suspect very virulent infectious bursal disease virus genomic RNA samples. 1609 30

Polymerase chain reaction (PCR)-based approaches to the detection, differentiation and characterization of avian pathogens continue to be developed and refined. The PCRs, or reverse transcriptase-PCRs, may be general, designed to detect all or most variants of a pathogen, or to be serotype, genotype or pathotype specific. Progress is being made with respect to making nucleic acid approaches more suitable for use in diagnostic laboratories. Robotic workstations are now available for extraction of nucleic acid from many samples in a short time, for routine diagnosis. Following general PCR, the DNA products are commonly analyzed by restriction endonuclease mapping (restriction fragment length polymorphism), using a small number of restriction endonucleases, based on a large body of sequence data. Increasingly, however, nucleotide sequencing is being used to analyze the DNA product, in part due to the expanding use of non-radioactive sequencing methods that are safe and enable high throughout. In this review, I highlight some recent developments with many avian viruses: Newcastle disease virus; circoviruses in canary and pigeon; infectious bursal disease virus (Gumboro disease virus); avian adenoviruses, including Angara disease/infectious hydropericardium virus, haemorrhagic enteritis virus of turkeys, and egg drop syndrome virus; avian herpesviruses, including infectious laryngotracheitis virus, duck plague virus, psittacine herpesvirus (Pacheco's parrot disease virus), Marek's disease virus and herpesvirus of turkeys; avian leukosis virus (associated with lymphoid leukosis or myeloid leukosis, and egg transmission); avian pneumoviruses (turkey rhinotracheitis virus); avian coronaviruses, including infectious bronchitis virus, turkey coronavirus and pheasant coronavirus; astrovirus, in the context of poult enteritis and mortality syndrome, and avian nephritis virus; and avian encephalomyelitis virus, a picornavirus related to hepatitis A virus.
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PMID:Innovation and discovery: the application of nucleic acid-based technology to avian virus detection and characterization. 1918 52

Infectious bursal disease virus (IBDV) is resistant to many environmental stresses and often persists on farms for months. This study investigated survival of a vaccine strain of IBDV in the bursa of Fabricius and splenic tissue from experimentally infected chickens and in splenic tissue and manure that had been inoculated with the virus. The specimens buried in compost were contained within nylon mesh bags, and the tissues were enclosed within the abdominal cavity of chicken carcasses. Extracts of composted specimens were inoculated into Vero cell cultures, and real-time reverse transcriptase PCR was used to quantify the virus in the cultures. By day 7 in compost, the temperature had been slightly above 55 C for 2.6 days and IBDV had been inactivated in specimens that had been inoculated with virus but had survived in tissues that had been taken from infected chickens. By day 14, the temperature had been above 55 C for 8.8 days and the virus was inactivated in all specimens. The results suggest that composting of poultry carcasses and manure would help to break the cycle of infection with IBDV and that the virus could be valuable as a surrogate for predicting the inactivation of less resistant viruses during composting.
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PMID:Infectious bursal disease virus as a surrogate for studies on survival of various poultry viruses in compost. 2060 40

Infectious bursal disease virus (IBDV) is endemic to most poultry-producing countries worldwide. Immunosuppressive classical and variant IBDV strains endemic to Australia are genetically distinct from other international strains. We report the results of infection experiments with Australian classical strain 06/95 and variant strain 02/95 in SPF chickens. We tested the effects of strain and age of infection on bursal atrophy, viral RNA (vRNA) load in bursa of Fabricius (bursa), spleen, thymus, caecal tonsils, faeces, litter and exhaust dust as determined by real-time reverse transcriptase polymerase chain reaction. The two IBDV strains did not differ in the degree of bursal atrophy induced, lymphoid organ distribution and faecal shedding but variant strain 02/95 induced a greater antibody response to the infection than classical strain 06/95 which was associated with a more rapid decline in IBDV vRNA genome copy number (VCN) in lymphoid organs and faeces. Infection at 14 days of age induced greater bursal atrophy and higher vRNA copy number in lymphoid tissues than infection on the day of hatching, indicating true age susceptibility independent of maternal antibody (Mab) status. The direction of the association between rankings for IBDV vRNA load in bursa and relative bursal weight changed from positive at 3 and 6 days post-infection to negative at 28 days post-infection. Intra-tracheal administration of dust collected from chickens infected with IBDV resulted in successful transmission of IBDV. IBDV vRNA was detected successfully at high levels in the environmental litter and dust samples.
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PMID:Pathogenicity, tissue distribution, shedding and environmental detection of two strains of IBDV following infection of chickens at 0 and 14 days of age. 2776

Infectious bursal disease virus (IBDV) contains two genome segments (segment A/segment B) that can reassort among the viruses. Reassortant IBDVs have been identified in several countries including the United States. These reassortant viruses usually include at least one genome segment from a very virulent (vv)IBDV strain. In vivo virulence of six reassortant IBDV from the United States was assessed relative to the virulence of three frequently described IBDV pathotypes: vvIBDV (rB strain), classic virulent (cv)IBDV (STC strain), and subclinical (sc)IBDV (Del-E strain). Morbidity and mortality in 4-wk-old specific-pathogen-free (SPF) leghorns indicated that reassortant IBDV with a vv genome segment A and non-vv segment B were less pathogenic than the vv/vv rB strain but more pathogenic than the cv/cv STC strain. The sc/vv IBDV strain D6337 (sc/vv) was comparable to the STC strain in pathogenicity. Viruses with a serotype 2 (ser2) genome segment A, regardless of the type of genome segment B, did not cause clinical disease in SPF chickens or turkeys. None of the reassorted viruses caused morbidity, mortality, or gross lesions in SPF turkeys. Histopathologic lesions in the bursa of turkeys were not observed in any group except those challenged with the serotype 2 OH strain, which had a mild lymphocytic depletion. No mortality was observed in maternally immune broilers inoculated with any of the IBDV pathotypes at 1, 2, 3, and 4 wk of age. No bursal lesions were observed in any of the broiler chicken groups at 1 wk of age except for the D2712 (ser2/cv)-inoculated birds that had mild lymphocyte depletion. Based on evaluation of bursal lesion scores and IBDV reverse transcriptase-PCR on broilers challenged at 2 wk of age, the K669 (vv/ser2) virus broke through the maternal immunity while the STC, Del-E, rB, D2712 (ser2/cv), and 7741 (vv/cv) viruses did not. All viruses broke through maternal immunity in the broilers at 3 wk of age except the Del-E scIBDV and D2712 (ser2/cv) reassortant IBDVs. At 4 wk of age, maternal antibodies were very low and bursal lesions were observed in all broilers challenged with the viruses. The data indicate that genome reassortant IBDVs are less pathogenic than is the rB (vv/vv) IBDV. However, the reassortant viruses with a vv genome segment A can still cause morbidity and mortality in SPF chickens, and they were able to break through maternal immunity produced via use of commercial classic and variant vaccines at an early age. This suggests that current breeder vaccination programs may not adequately protect against the reassortant vv/ser2 and vv/cv IBDV strains.
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PMID:Pathogenicity of Genome Reassortant Infectious Bursal Disease Viruses in Chickens and Turkeys. 2790 5

Infectious bursal disease (IBD) is an immunosuppressive, acute and highly contagious illness of growing-poultry stock infected with infectious bursal disease virus (IBDV). It is common in Pakistan, causing potential economic losses throughout the year. The objective of the study is to propose a rapid, sensitive and specific diagnostic tool, and compare it with existing commonly used reverse transcriptase polymerase chain reaction (RT-PCR) method for IBDV. Different primers were used for RT-PCR and reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) to target the IBD virus. RT-LAMP primers showed prodigious specificity without cross reaction to the other animal pathogens. Moreover, RT-LAMP was found to have 10 times higher selectivity for IBDV identification as compared to RT-PCR. RT-LAMP detected 9.2% more field samples than RT-PCR. Sequences of PCR products were determined and phylogenetic analysis of research isolates revealed its maximum similarity with indigenous and Indian IBDV isolates. RT-LAMP was found to be simple, specific, less laborious and a better technique as compared to RT-PCR for quick analysis. In general, RT-LAMP was declared positive on observing turbidity or adding fluorescence staining reagent such as SYBR Green I. The options of direct use of field sample homogenate and viewing directly the peaks in the graph shown on a monitor/laptop have made it much more convenient and time saving than gel based RT-PCR.
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PMID:Rapid detection of infectious bursal disease by loop-mediated isothermal amplification for field analysis. 3004 20


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