Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.7.49 (reverse transcriptase)
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Telomeres are the terminal portions of chromosomes and consist of the repeated nucleotide sequence TTAGGG. Chromosomes lose a small amount of telomeric deoxyribonucleic acid (DNA) after each cell replication. A hypothetical function of telomeric DNA is to allow for a finite number of cell divisions without loss of functional genes. A second proposed function of telomeric DNA is to prevent undesirable interactions between chromosomal ends and cellular repair enzymes. In cells that maintain a proliferative capacity, such as stem cells and cancer cells, telomere length is maintained by the reverse transcriptase, telomerase. Virtually every major human malignancy has been evaluated for telomerase activity and approximately 80% to 90% have demonstrated the presence of telomerase. In this article we review the current assays available for telomerase detection and discuss their relative strengths and limitations.
Conn Med 2001 Nov
PMID:Measuring telomerase activity for the early detection of cancer. 1176 50

An infectious bronchitis virus (IBV) was isolated from commercial broilers from the state of California exhibiting respiratory distress, inflamed tracheas, airsaculitis, and edematous lungs. After reverse transcriptase-polymerase chain reaction (RT-PCR), the California isolate exhibited an identical restriction fragment length polymorphism (RFLP) pattern to some isolates obtained from California, known as California 99 isolates. Commercial Mass-Conn and Mass-Ark vaccines were used to vaccinate commercial broiler chickens via eye drop once at 1 or 10 days of age or twice at 1 and 10 days of age. At 27 days of age the birds were challenged via eye drop with the isolated IBV California 99 strain. Protection was measured by failure to reisolate the challenge virus from tracheas 5 days postchallenge and complemented withthe tracheal and epithelium thickness scores. When the Mass-Ark vaccine was included in the vaccination programs, there was protection against challenge with the IBV California 99 isolate. The Mass-Conn vaccine conferred protection when used once at 1 day of age and twice at 1 and 10 days of age. However, no total protection was achieved when used as the only vaccine at 10 days of age, since one of the replicates was positive for virus isolation. Significant differences (P < 0.05) in the epithelium thickness and tracheal scores were observed between the unvaccinated-unchallenged group and the groups vaccinated once or twice with the Mass-Conn vaccine. Based on these results, all chickens were protected against the California 99 isolate when the IBV Arkansas type was used as a vaccine.
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PMID:Evaluation of the protection conferred by commercial vaccines against the California 99 isolate of infectious bronchitis virus. 1470 75

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

Real-time quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) assays have been used to detect the presence of challenge virus when the efficacy of infectious bronchitis virus (IBV) vaccine against field viruses is being experimentally evaluated. However, federal guidelines for licensing IBV vaccines indicate that challenge-virus detection following vaccination is to be conducted in embryonated eggs. In this study, we examined qRT-PCR data with the use of universal and type-specific primers and probe sets for IBV detection and compared those data with challenge-virus detection in embryonated eggs to determine if the two methods of evaluating vaccine efficacy are comparable. In addition, we tested the qRT-PCR assays on thermocyclers from two different manufacturers. We found the universal IBV primers and probe set to be comparable to challenge-virus detection in embryonated eggs. However, for some IBV types (Mass41 and Conn on the SmartCycler II and Ark, Mass41, Conn, and GA98 on the ABI 7500) the qRT-PCR assay was more sensitive than virus detection in embryonated eggs. This may simply be due to the universal IBV qRT-PCR assay being more sensitive than virus detection in eggs or to the assay detecting nucleic acid from nonviable virus. This finding is important and needs to be considered when evaluating challenge-virus detection for vaccination and challenge studies, because qRT-PCR could potentially identify positive birds that would otherwise be negative by virus detection in embryonated eggs; thus it could lead to a more stringent measure of vaccine efficacy. We also found that the IBV type-specific primers and probe sets designed in this study were in general less sensitive than the universal IBV primers and probe set. Only the Ark-DPI-spedcific assay on the SmartCycler II and the Ark-DPI-, Mass41-, and DE072/GA98- (for detection of GA98 virus only) specific assays on the ABI 7500 were comparable in sensitivity to virus detection in eggs. We found that a number of variables, including the virus type examined, primers and probe efficiency and stability, and assay conditions, including thermocycler platform, can affect the data obtained from qRT-PCR assays. These results indicate that qRT-PCR assays can be used to detect IBV challenge virus, but each assay, including the assay conditions and thermocycler, should be individually evaluated if those data are expected to be comparable to virus detection in embryonated eggs.
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PMID:Detection of infectious bronchitis virus with the use of real-time quantitative reverse transcriptase-PCR and correlation with virus detection in embryonated eggs. 2551 34