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)

We investigated the feasibility of using Flinders Technology Associates (FTA) filter cards for the storage of allantoic fluid containing an infectious bronchitis virus (IBV), such as Arkansas-DPI, Connecticut, and Massachusetts, and for their identification by reverse transcriptase (RT)-polymerase chain reaction (PCR) and characterization by restriction fragment length polymorphism (RFLP) or nucleotide sequencing. FTA paper is a cotton-based cellulose membrane containing lyophilized chemicals that lyses many types of bacteria and viruses. IBV was inactivated upon contact with the FTA, as shown by the inability of the virus to be propagated in embryonating chicken eggs. RT-PCR of the S1 gene showed that viral RNA in allantoic fluid remained stable after storage on FTA filter cards and that the stability was time and temperature sensitive for the large (1700 base pair [bp]) but not the small (383 bp) PCR products. Analysis of the amplified products showed that molecular characterization is feasible in allantoic fluid stored on FTA under nonfavorable environmental conditions (41 C) for at least 15 days. The use of FTA cards for the collection, transport, and storage of IBV-containing samples is safe, inexpensive, and adequate for molecular diagnosis. We propose that specimens coming from overseas on FTA cards would be first analyzed by RT-PCR with primers yielding a 1700-bp product followed by RFLP of the positive cases. Negative cases would be analyzed with primers yielding a 383-bp product (to exdude detrimental effect of the storage conditions) followed by nucleotide sequencing of the positive cases.
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PMID:Molecular detection and serotyping of infectious bronchitis virus from FTA filter paper. 1583 8

Fourteen infectious bronchitis viruses (IBVs) were isolated in Korea between 2001 and 2003 from chickens suspected to be infected with IBVs. The nucleocapsid (N) protein genes of the various IBVs were amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) and were cloned and sequenced, and the nucleotide and deduced amino acid sequences were compared with published sequences for non-Korean IBV strains. The Korean IBV isolates shared amino acid sequence similarity of between 89.2% (K203-02 and K1255-03) and 98.3% (K434-01 and K281-01) with each other and exhibited amino acid sequence similarity between 57.0% (K774-01 and V18/91) and 96.6% (K507-01 and JP8147) with non-Korean IBV strains. Phylogenetic analysis of the deduced N protein amino acid sequences resulted in the segregation of Korean IBV isolates into three different clusters, with cluster assignments differing for some of the isolates from those obtained with analysis of the S1 glycoprotein. Korean IBV isolates K069-01, K281-01, K434-01, K504-01, K774-01, K748-01, K044-02, K058-02, K161-02, K203-02, and K234-02 formed an independent cluster comprised only of Korean IBV isolates. Another Korean IBV isolate, K210-02, belonged to a cluster that included IBV strains isolated in USA, the Netherlands and China. Recent Korean IBV isolates K514-03 and K1255-03 grouped into a third distinct cluster related to a Chinese IBV strain. As deduced from phylogenetic analysis, some IBV isolates appear to have arisen from the recombination of IBV strains with different origins.
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PMID:Variations in the nucleocapsid protein gene of infectious bronchitis viruses isolated in Korea. 1602 40

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

In 1993, a new molecular typing method for infectious bronchitis virus (IBV) was introduced. This method uses reverse transcriptase-polymerase chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP) analysis of the spike gene to obtain RFLP patterns that correlate with serotype. Using that test at the Poultry Diagnostic and Research Center (PDRC, University of Georgia, Athens, GA), we have identified a total of 1523 IBV isolates in the past 11 yr. The data were obtained from clinical samples submitted to our laboratory from birds with clinical signs characteristic of IBV infection. The samples are primarily from the southeastern United States but are also from many other states as well as from outside the United States. Most of the isolations occurred during July, followed by May, April, November, October, and January. The fewest number of isolates identified on an annual basis was 20 in 2003. An unusually high number of isolations occurred in 1997 (318 isolations) and 1999 (246 isolations), which coincided with the GAV variant virus and GA98 variant virus outbreaks respectively. By far, the Ark-DPI strain was the most frequently identified type of IBV and ranged from 23% to 65% of total isolations per year. Ark-like isolates, defined as having a similar but unique RFLP pattern from the Ark-DPI vaccine strain were identified every year of the study except in 1996. In addition, new Ark-like isolates continued to emerge each year (except in the year 2000) beginning in 1997, reflecting the ability of that IBV type to undergo genetic drift. Eighty-two different variant viruses were identified although only two (GAV and GA98) became persistent and caused widespread disease. Some viruses tended to be geographically restricted to a given area (CAV in California and MX97-8147 in Mexico), whereas others were widespread (Ark-DPI, Conn, DE072, and Mass). The Florida, Gray, Holte, Iowa, and JMK types were not detected during the 11-yr period, and no foreign virus types were detected in the United States. These data show that IBV variant viruses are consistently circulating in commercial poultry and are capable of causing disease outbreaks. Our observations highlight the importance of constantly monitoring IBV as well as other coronaviruses like severe acute respiratory syndrome-coronavirus that have the ability to change and emerge to cause disease in a susceptible host.
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PMID:Data from 11 years of molecular typing infectious bronchitis virus field isolates. 1640 10

The number of avian species in which coronaviruses have been detected has doubled in the past couple of years. While the coronaviruses in these species have all been in coronavirus Group 3, as for the better known coronaviruses of the domestic fowl (infectious bronchitis virus [IBV], in Gallus gallus), turkey (Meleagris gallopavo) and pheasant (Phasianus colchicus), there is experimental evidence to suggest that birds are not limited to infection with Group 3 coronaviruses. In China coronaviruses have been isolated from peafowl (Pavo), guinea fowl (Numida meleagris; also isolated in Brazil), partridge (Alectoris) and also from a non-gallinaceous bird, the teal (Anas), all of which were being reared in the vicinity of domestic fowl. These viruses were closely related in genome organization and in gene sequences to IBV. Indeed, gene sequencing and experimental infection of chickens indicated that the peafowl isolate was the H120 IB vaccine strain, while the teal isolate was possibly a field strain of a nephropathogenic IBV. Thus the host range of IBV does extend beyond the chicken. Most recently, Group 3 coronaviruses have been detected in greylag goose (Anser anser), mallard duck (Anas platyrhynchos) and pigeon (Columbia livia). It is clear from the partial genome sequencing of these viruses that they are not IBV, as they have two additional small genes near the 3' end of the genome. Twenty years ago a coronavirus was isolated after inoculation of mice with tissue from the coastal shearwater (Puffinus puffinus). While it is not certain whether the virus was actually from the shearwater or from the mice, recent experiments have shown that bovine coronavirus (a Group 2 coronavirus) can infect and also cause enteric disease in turkeys. Experiments with some Group 1 coronaviruses (all from mammals, to date) have shown that they are not limited to replicating or causing disease in a single host. SARS-coronavirus has a wide host range. Clearly there is the potential for the emergence of new coronavirus diseases in domestic birds, from both avian and mammalian sources. Modest sequence conservation within gene 1 has enabled the design of oligonucleotide primers for use in diagnostic reverse transcriptase-polymerase chain reactions, which will be useful for the detection of new coronaviruses.
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PMID:Coronaviruses in poultry and other birds. 1653 57

A virus (AV71/99) was isolated from a green-cheeked Amazon parrot by propagation and passage in both primary embryo liver cells derived from blue and yellow macaw (Ara ararauna) embryos and chicken embryo liver cells. Electron microscopic examination of cytopathic agents derived from both types of cell cultures suggested that it was a coronavirus. This was confirmed using a pan-coronavirus reverse transcriptase polymerase chain reaction that amplified part of gene 1 that encodes the RNA-dependent RNA polymerase. The deduced sequence of 66 amino acids had 66 to 74% amino acid identity with the corresponding sequence of coronaviruses in groups 1, 2 and 3. Several other oligonucleotide primer pairs that give PCR products corresponding to genes 3, 5, N and the 3'-untranslated region of infectious bronchitis virus, turkey coronavirus and pheasant coronavirus (all in group 3) failed to do so with RNA from the parrot coronavirus. This is the first demonstration of a coronavirus in a psittacine species.
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PMID:Isolation of a coronavirus from a green-cheeked Amazon parrot (Amazon viridigenalis Cassin). 1659 4

Human rhinoviruses (HRV), members of the Picornaviridae family, are comprised of over 100 different virus serotypes. HRV represent the single most important etiological agents of the common cold [Arruda, E., Pitkaranta, A., Witek Jr., T.J., Doyle, C.A., Hayden, F.G., 1997. Frequency and natural history of rhinovirus infections in adults during autumn. J. Clin. Microbiol. 35, 2864-2868; Couch, R.B., 1990. Rhinoviruses. In: Fields, B.N., Knipe, D.M. (Eds.), Virology. Raven Press, New York, pp. 607-629; Turner, R.B., 2001. The treatment of rhinovirus infections: progress and potential. Antivir. Res. 49 (1), 1-14]. Although HRV-induced upper respiratory illness is often mild and self-limiting, the socioeconomic impact caused by missed school or work is enormous and the degree of inappropriate antibiotic use is significant. It has been estimated that upper respiratory disease accounts for at least 25 million absences from work and 23 million absences of school annually in the United States [Anzueto, A., Niederman, M.S., 2003. Diagnosis and treatment of rhinovirus respiratory infections. Chest 123 (5), 1664-1672; Rotbart, H.A., 2002. Treatment of picornavirus infections. Antivir. Res. 53, 83-98]. Increasing evidences also describe the link between HRV infection and more serious medical complications. HRV-induced colds are the important predisposing factors to acute otitis media, sinusitis, and are the major factors in the induction of exacerbations of asthma in adults and children. HRV infections are also associated with lower respiratory tract syndromes in individuals with cystic fibrosis, bronchitis, and other underlying respiratory disorders [Anzueto, A., Niederman, M.S., 2003. Diagnosis and treatment of rhinovirus respiratory infections. Chest 123 (5), 1664-1672; Gern, J.E., Busse, W.W., 1999. Association of rhinovirus infections with asthma. Clin. Microbiol. Rev. 12 (1), 9-18; Pitkaranta, A., Arruda, E., Malmberg, H., Hayden, F.G., 1997. Detection of rhinovirus in sinus brushings of patients with acute community-acquired sinusitis by reverse transcription-PCR. J. Clin. Microbiol. 35, 1791-1793; Pitkaranta, A., Virolainen, A., Jero, J., Arruda, E., Hayden, F.G., 1998. Detection of rhinovirus, respiratory syncytial virus, and coronavirus infections in acute otitis media by reverse transcriptase polymerase chain reaction. Pediatrics 102, 291-295; Rotbart, H.A., 2002. Treatment of picornavirus infections. Antivir. Res. 53, 83-98]. To date, no effective antiviral therapies have been approved for either the prevention or treatment of diseases caused by HRV infection. Thus, there still exists a significant unmet medical need to find agents that can shorten the duration of HRV-induced illness, lessen the severity of symptoms, minimize secondary bacterial infections and exacerbations of underlying disease and reduce virus transmission. Although effective over-the-counter products have been described that alleviate symptoms associated with the common cold [Anzueto, A., Niederman, M.S., 2003. Diagnosis and treatment of rhinovirus respiratory infections. Chest 123 (5), 1664-1672; Gwaltney, J.M., 2002a. Viral respiratory infection therapy: historical perspectives and current trials. Am. J. Med. 22 (112 Suppl. 6A), 33S-41S; Turner, R.B., 2001. The treatment of rhinovirus infections: progress and potential. Antivir. Res. 49 (1), 1-14; Sperber, S.J., Hayden, F.G., 1988. Chemotherapy of rhinovirus colds. Antimicrob. Agents Chemother. 32, 409-419], this review will primarily focus on the discovery and development of those agents that directly or indirectly impact virus replication specifically highlighting new advances and/or specific challenges with their development.
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PMID:Rhinovirus chemotherapy. 1667 37

The objective of this study was to compare the presence of the Arkansas (Ark) and Massachusetts (Mass) serotypes of infectious bronchitis virus (IBV) in the tracheas and cecal tonsils of commercial broilers after vaccination at 1 day of age by coarse spray. When given as a single serotype vaccine, the Mass strain was detected by reverse transcriptase-polymerase chain reaction (RT-PCR)-restriction fragment length polymorphism (RFLP) only in the tracheas, whereas the Ark strain was detected in both the tracheas and cecal tonsils. By in situ hybridization, the Mass and Ark nucleocapsid (Nc) genes were detected only at 7 days in the tracheas. When both strains were given in the mixed vaccine, the Mass strain was more consistently detected by RT-PCR-RFLP in the tracheas and cecal tonsils at early stages of infection (up to 14 days) and the Arkansas strain was more consistently detected at late stages of infection (21 and 28 days). By in situ hybridization, the IBV Nc gene was more consistently detected in the trachea at early stages of infection (7, 14, and 21 days) and in the cecal tonsils at late stages of infection (21, 28, and 35 days). In general, the Mass strain was more frequently recovered from the tracheal and cecal tonsil tissues at earlier stages of infection and the Ark strain was recovered at later stages of infection.
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PMID:Detection of Massachusetts and Arkansas serotypes of infectious bronchitis virus in broilers. 1686 85

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

Molecular characterization of infectious bronchitis viruses (IBVs) isolated between 1998 and 2002 from chickens in Russia was performed. More than 250 field samples were tested by reverse transcriptase-polymerase chain reaction using two sets of primers corresponding to the most conserved 3'-untranslated region and the most variable S1 gene region of the viral genome. Ninety-one IBV isolates were characterized by phylogenetic analysis of the S1 gene hypervariable region comprising 136 to 558 nucleotides. The major group of isolates (38 viruses) showed very close sequence relationship with strains of the Massachusetts genotype circulating in Russia since the early 1970s. The analysed region of the other 22 Russian IBVs was similar (from 89 to 98% identity) to that from the strains of European genotypes including D274 (nine isolates), 793/B (10 isolates), and B1648, 624/I and Italy-02 (one isolate in each group). Two isolates from very distant geographic locations in Russia (Far East and the European part) clustered together with Chinese strains of QXIBV genotype. None of the remaining 27 Russian isolates showed a close sequence relationship with known IBV strains available in sequence databases. The majority of these variant viruses clustered into the six novel Russian genotypes, often correlating with their geographic location. The remaining five of them were placed outside these unique groups, also representing new genotypes. These data for the first time demonstrated the high genetic diversity of IBV isolates circulating in Russia.
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PMID:Molecular epizootiology of avian infectious bronchitis in Russia. 1699 Jan 48


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