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Target Concepts:
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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)
Competitive quantitative PCRs were used to examine the consequences of stereotactically injecting a highly attenuated herpes simplex virus type 1 mutant into rat brains. This mutant virus, designated RR1CAT/RR2lacZ, was engineered so that coding sequences of the genes UL39 and UL40 specifying the subunits of the viral ribonucleotide reductase were replaced by the chloramphenicol acetyltransferase (CAT) and the lacZ gene coding sequences, respectively. Stereotactic injection of this virus into the hippocampal region of the rat brain resulted in a
localized infection
. Viral gene products were visualized by immunochemical, cytochemical, or in situ hybridization techniques in the injected hippocampal region at 2 days postinjection. Viral genomes, represented by glycoprotein B (gB), latency-associated transcript (LAT), and lacZ sequences could be amplified by PCR from templates obtained by scraping hippocampal tissue off single 10-microns frozen sections. Both gB message and LAT could be detected by
reverse transcriptase
(RT)-PCR. At day 7 postinjection, neither CAT message, gB message, nor beta-galactosidase activity could be visualized by the same techniques, although viral DNA was detected by PCR and LAT could be detected by RT-PCR. A similar pattern was seen at 8 weeks, suggesting that latency was established by the mutant virus in cells of the injected hippocampus. By competitive quantitative PCR, hippocampal sections were determined to contain 2.6 x 10(5) genome equivalents (represented by the gB gene) on day 2, 6.2 x 10(4) on day 7, and 8.3 x 10(4) at 8 weeks. By competitive quantitative RT-PCR, the numbers of LAT molecules at the same time points were 3.2 x 10(6), 1.3 x 10(6), and 1.2 x 10(6), respectively. The numbers of LAT molecules per genome equivalent were 12.5, 20.3, and 14.5, respectively, being approximately the same for each of the three time points. The data permit the conclusion that the RR mutant virus establishes latency in the rat brain with the persistence of the viral genome and the production of LAT molecules. Once latency is established, the numbers of viral genomes and LAT RNA molecules remain constant. Thus the competitive quantitative PCR and RT-PCR techniques provide very sensitive and reliable methods to quantitate viral DNA and RNA present in infected tissue.
...
PMID:Competitive quantitative PCR analysis of herpes simplex virus type 1 DNA and latency-associated transcript RNA in latently infected cells of the rat brain. 810 47
We present a prospective case-control study of 27 serologically confirmed dengue hemorrhagic fever (DHF) patients with severe central nervous system symptoms. Dengue associated encephalopathy accounted for 0.5% of 5,400 patients admitted with DHF. The mortality rate among children with encephalopathy was 22%, with the survivors experiencing a complete recovery. Liver enzymes and bilirubin were significantly elevated in the study group. In analysis of the cerebrospinal fluid (CSF),
reverse transcriptase
-polymerase chain reaction revealed dengue-3-specific RNA in one evaluated case. Dengue-specific immunoglobulin M was detected in CSF in 14 of 22 assessable patients, indicating a
localized infection
. Magnetic resonance imaging scans showed cerebral edema in the majority of patients, although encephalitis-like changes were less common. There was an equal distribution of primary and secondary infections. On the basis of previous reports and of the findings of our study, DHF probably encompasses an expanding clinical spectrum that infrequently involves encephalitis due to a direct neurotropic effect of dengue virus.
...
PMID:Prospective case-control study of encephalopathy in children with dengue hemorrhagic fever. 1179 85