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)

Biological transmission of arboviruses to a vertebrate host occurs when virions are expelled along with saliva during blood feeding by a hematophagous arthropod. We undertook experiments to determine whether mosquitoes expectorate flaviviruses in their saliva while sugar feeding. Batches of Culex annulirostris Skuse and Culex gelidus Theobald (Diptera: Culicidae) were orally infected with Japanese encephalitis (family Flaviviridae, genus Flavivirus, JEV), Kunjin (family Flaviviridae, genus Flavivirus, KUNV; a subtype of West Nile virus), and Murray Valley encephalitis (family Flaviviridae, genus Flavivirus, MVEV) viruses. After a 7-d extrinsic incubation, these mosquitoes were offered sucrose meals via cotton pledgets, which were removed daily and processed for viral RNA by using real-time TaqMan reverse transcriptase-polymerase chain reaction (RT-PCR) assays. JEV, MVEV, and KUNV RNA was detected in all pledgets removed from batches of Cx. gelidus on days 7-14 postexposure. In contrast, detection rates were variable for Cx. annulirostris, with KUNV detected in 0.3 M sucrose pledgets on all days postexposure, and JEV and MVEV detected on 57 and 50% of days postexposure, respectively. Higher concentrations of sucrose in the pledget did not increase virus detection rates. When individual JEV-infected Cx. gelidus were exposed to the sucrose pledget, 73% of mosquitoes expectorated virus with titers that were detectable by TaqMan RT-PCR. These results clearly show that flaviviruses are expectorated by infected mosquitoes during the process of sugar feeding on artificial pledgets. Potential applications of the method for arboviral bioassays and field surveillance are discussed.
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PMID:Expectoration of Flaviviruses during sugar feeding by mosquitoes (Diptera: Culicidae). 1791 18

We collected a total of 15,329 mosquitoes during weekly sampling in Davis, CA, from April through mid-October 2006 at 21 trap sites uniformly spaced 1.5 km apart over an area of approximately 26 km(2). Of these mosquitoes, 1,355 pools of Culex spp. were tested by multiplex reverse transcriptase-polymerase chain reaction, of which 16 pools (1.2%) were positive for West Nile virus (WNV). A degree-day model with a developmental threshold of 14.3 degrees C accurately predicted episodic WNV transmission after three extrinsic incubation periods after initial detection. Kriging interpolation delineated that Culex tarsalis were most abundant at traps near surrounding agriculture, whereas Cx. pipiens clustered within residential areas and greenbelt systems in the old portion of Davis. Spatial-temporal analyses were performed to test for clustering of locations of WNV-infected dead birds and traps with WNV-positive Cx. tarsalis and Cx. pipiens; human case incidence was mapped by census blocks. Significant multivariate spatial-temporal clustering was detected among WNV-infected dead birds and WNV-positive Cx. tarsalis, and a WNV-positive Cx. pipiens cluster overlapped areas with high incidences of confirmed human cases. Spatial analyses of WNV surveillance data may be an effective method to identify areas with an increased risk for human infection and to target control efforts to reduce the incidence of human disease.
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PMID:Risk factors associated with human infection during the 2006 West Nile virus outbreak in Davis, a residential community in northern California. 1818 85

We conducted a laboratory evaluation of the ability of commercial antigen-capture assays, the Rapid Analyte Measurement Platform (RAMP) and the VecTest wicking assay, as well as Real Time reverse transcriptase polymerase chain reaction (RT-PCR, Taqman) and Vero cell plaque assay to detect West Nile virus (WNV) in large mosquito pools. Real-Time PCR (Taqman) was the most sensitive, detecting WNV ribonucleic acid (RNA) in 100% of samples containing a single infected mosquito in pool sizes of up to 500 mosquitoes. Mosquito body tissues minimally impacted the ability of Real Time RT-PCR to detect WNV in a pool size of 500, reducing sensitivity by 0.6 log10 plaque-forming units (PFU)/ml. Vero cell plaque assay detected live virus from a single infected mosquito in 100% of pools containing up to 200 mosquitoes, but was unreliable at larger pool sizes. VecTest detected 100% of positive pools containing 50 mosquitoes with 5.8 log10 PFU/ml virus, 100 mosquitoes with 5.9 log10 PFU/ml, and 200 mosquitoes with 5.2 log10 PFU/ ml. The RAMP assay detected 100% of positive pools containing 50 mosquitoes with 3.3 log10 PFU/ml virus, 100 mosquitoes with 3.7 log10 PFU/ml, and 200 mosquitoes with 4.0 log10 PFU/ml. Results indicate that WNV can be reliably detected by all 4 assays in pools of mosquitoes exceeding 50 specimens, though there is some loss of sensitivity with very large pool sizes.
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PMID:Detection of West Nile virus in large pools of mosquitoes. 1824 May 15

West Nile virus surveillance was conducted at five sites in New Castle County, DE, and one site in Salem County, NJ, from June through September, 2004, using dry ice-baited Centers for Disease Control miniature light traps, infusion-baited gravid traps, and resting boxes. All trap types were simultaneously placed at each site every two weeks and run overnight. Collected mosquitoes were identified to species, pooled, and analyzed for virus using a real-time reverse transcriptase polymerase chain reaction test. In total, 47,972 mosquitoes in 29 species or species groups were analyzed. Light traps collected 60,201 mosquitoes in 28 species or species groups. Gravid traps collected 3,195 mosquitoes in 19 species or species groups. Resting boxes collected 99 mosquitoes in nine species or species groups. In total, 1,500 mosquito pools were tested for WNV resulting in ten positive pools. All positive pools consisted of Culex pipiens, Culex restuans, or Culex salinarius. Seven positive pools were from gravid traps and three were from light traps despite testing almost 14 times as many pools from light traps. The overall infection rate from gravid traps was nearly 33 times greater than the infection rate from light traps, 2.29 and 0.07 infected mosquitoes per 1,000, respectively. The results demonstrate the advantage of using gravid traps for West Nile virus surveillance over light traps or resting boxes.
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PMID:Comparison of light traps, gravid traps, and resting boxes for West Nile virus surveillance. 1826 May 19

The prevalence of West Nile virus (WNV) was determined in mosquitoes between November 2002 and October 2004 in East Baton Rouge Parish, LA. A total of 244,374 female mosquitoes were collected and tested by viral isolation. Additionally, 131,896 female mosquitoes were collected in 2003 and tested by VecTest and 167,175 female mosquitoes were collected in 2004 and tested by reverse transcriptase-polymerase chain reaction (RT-PCR). West Nile virus was isolated by cell culture from 17 (47.2%) out of 36 mosquito species collected over the study period. In 2003, WNV was detected in 9 (33.3%) out of 27 species tested by VecTest. In 2004, 14 (50%) out of the 28 mosquito species tested by RT-PCR were positive for WNV. The species with the greatest number of WNV-positive pools detected by all 3 testing methods was Culex quinquefasciatus. A significantly greater proportion of Cx. salinarius pools collected in light traps placed at a 3-m height were positive for WNV by viral isolation than in pools collected in light traps placed at a 1.5-m height.
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PMID:West Nile virus detection in mosquitoes in East Baton Rouge Parish, Louisiana, from November 2002 to October 2004. 1843 11

Nonstructural protein 4 (NSP4), encoded by group A rotavirus genome segment 10, is a multifunctional protein and the first recognized virus-encoded enterotoxin. The NSP4 gene has been sequenced, and five distinct genetic groups have been described: genotypes A-E. NSP4 genotypes A, B, and C have been detected in humans. In this study, the NSP4-encoding gene of human rotavirus strains of different G and P genotypes collected from children between 1987 and 2003 in three cities of West Central region of Brazil was characterized. NSP4 gene of 153 rotavirus-positive fecal samples was amplified by reverse transcriptase-polymerase chain reaction and then sequenced. For phylogenetic analysis, NSP4 nucleotide sequences of these samples were compared to nucleotide sequences of reference strains available in GenBank. Two distinct NSP4 genotypes could be identified: 141 (92.2%) sequences clustered with NSP4 genotype B, and 12 sequences (7.8%) clustered with NSP4 genotype A. These results reinforce that further investigations are needed to assess the validity of NSP4 as a suitable target for epidemiologic surveillance of rotavirus infections and vaccine development.
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PMID:Molecular characterization of the NSP4 gene of human group A rotavirus samples from the West Central region of Brazil. 1859 1

We analysed the sequence diversity in the reverse transcriptase (RT)/ribonuclease H (RNaseH) coding region of 19 badnavirus isolates infecting yam (Dioscorea spp.) in Ghana, Togo, Benin, and Nigeria. Phylogenetic analysis of the deduced amino acid sequences revealed that the isolates are broadly divided into two distinct species, each clustering with Dioscorea alata bacilliform virus (DaBV) and Dioscorea sansibarensis bacilliform virus (DsBV). Fourteen isolates had 90-96% amino acid identity with DaBV, while four isolates had 83-84% amino acid identity with DsBV. One isolate from Benin, BN4Dr, was distinct and had 77 and 75% amino acid identity with DaBV and DsBV, respectively, and may be a member of a new badnavirus species infecting yam in West Africa. Viruses of the two main species were present in Ghana, Togo and Benin and were observed to infect both D. alata and D. rotundata indiscriminately. This is the first confirmed report of DsBV infection in yam in Ghana and Togo. The results of this study demonstrate that members of two distinct species of badnaviruses infect yam in the West African yam zone and suggest a putative new species, BN4Dr. We also conclude that these species are not confined to limited geographic regions or specific for yam host species. However, the three badnavirus species are serologically related. The sequence information obtained from this study can be used to develop PCR-based diagnostics to detect members of the various species and/or strains of badnaviruses infecting yam in West Africa.
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PMID:Sequence diversity among badnavirus isolates infecting yam (Dioscorea spp.) in Ghana, Togo, Benin and Nigeria. 1903 Sep 55

West Nile virus (WNV) is known to affect captive populations of alligators and, in some instances, cause significant mortalities. Alligators have been shown to amplify the virus, serve as a reservoir host, and even represent a source of infection for humans. This study describes a cutaneous manifestation of WNV in captive-reared American alligators (Alligator mississippiensis), previously described as lymphohistiocytic proliferative syndrome of alligators (LPSA), based on the findings of gross examination, histopathologic evaluation, WNV antibody testing, and WNV reverse transcriptase polymerase chain reaction (RT-PCR). Forty alligators with LPSA and 41 controls were examined. There was a significant difference (P = 0.01(-21)) in the WNV serostatus between the treatment group (100%) and the control group (0%, 95% CI: 0-7.3%). In the treatment group, 97.5% (39/40) (95% CI: 92.7-102.3%) of the LPSA skin lesions were positive for WNV via RT-PCR. Of the skin sections within the treatment group that had no LPSA lesions, 7.5% (3/40) (95% CI: 0-15.7%) were positive for WNV. In the control group, all of the skin samples were negative for WNV (41/41) (0%; 95% CI: 0-7.3%). The LPSA skin lesions were significantly more likely to be WNV positive by RT-PCR when compared to control animals (P = 0.07(-20)) and normal skin sections from affected animals (P = 0.08(-16)). There was no significant difference in the WNV RT-PCR results between control animals and normal skin sections from affected animals (P = 0.24). These findings suggest that LPSA is a cutaneous manifestation of WNV in alligators.
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PMID:Association of West Nile virus with lymphohistiocytic proliferative cutaneous lesions in American alligators (Alligator mississippiensis) detected by RT-PCR. 1911 Jun 97

We sought to elucidate the role of migratory birds in transmission of H5N1 in an enzoonotic area. Resident, captive, and migratory birds were sampled at five sites in Java, Indonesia. Mist nets were used to trap birds. Birds were identified to species. RNA was extracted from swabs and reverse transcriptase polymerase chain reaction (RT-PCR) conducted for the HA and M genes of H5N1. Antibodies were detected by enzyme-linked immunosorbent assay and hemagglutination inhibition test. Between October 2006 and September 2007, a total of 4,067 captive, resident, and migratory birds comprising 98 species in 23 genera were sampled. The most commonly collected birds were the common sandpiper (6% of total), striated heron (3%), and the domestic chicken (14%). The overall prevalence of H5N1 antibodies was 5.3%. A significantly higher percentage of captive birds (16.1%) showed antibody evidence of H5N1 exposure when compared to migratory or resident birds. The greatest number of seropositive birds in each category were Muschovy duck (captive), striated heron (resident), and the Pacific golden plover (migratory). Seven apparently well captive birds yielded molecular evidence of H5N1 infection. Following amplification, the HA, NA, and M genes were analyzed. Phylogenetic analysis of the HA gene showed that the isolates were 97% similar to EU124153.1 A/chicken/West Java/Garut May 2006, an isolate obtained in a similar region of West Java. While no known markers of neuraminidase inhibitor resistance were found within the NA gene, M segment analysis revealed the V27A mutation known to confer resistance to adamantanes. Our results demonstrate moderate serologic evidence of H5N1 infection in captive birds, sampled in five sites in Java, Indonesia, but only occasional infection in resident and migratory birds. These data imply that in an enzoonotic region of Indonesia the role of migratory birds in transmission of H5N1 is limited.
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PMID:H5N1 surveillance in migratory birds in Java, Indonesia. 1927 96

West Nile virus (WNV) infection in humans can cause neurological deficits, including flaccid paralysis, encephalitis, meningitis, and mental status change. To better understand the neuropathogenesis of WNV in the peripheral and the central nervous systems (PNS and CNS), we used a mouse footpad inoculation model to simulate a natural peripheral infection. Localization of WNV in the nervous system using this model has suggested two routes of viral invasion of the CNS: axonal retrograde transport (ART) from the PNS and hematogenous diffusion via a breakdown in the blood-choroid-plexus barrier. C57BL/6J mice were treated with nocodazole, a microtubule inhibitor that blocks ART, prior to infection with WNV. Nocodazole-treated WNV-infected mice developed a viremia 1.5 log(10) greater than untreated WNV-infected control mice at days 3 to 4 post infection (PI). Although viremia was greater in nocodazole-treated mice, detection of virus in brain tissue (spinal cord, cortex, brainstem, and cerebellum), as measured by real-time reverse transcriptase-polymerase chain reaction (RT-PCR), did not occur until day 7. At these later time points (7 and 9 days PI), nocodazole-treated WNV-infected animals attained viral titers in these tissues similar to titers in the untreated WNV-infected control animals. These results demonstrate that a single dose of nocodazole delays, but does not block, WNV infection of the brain.
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PMID:Nocodazole delays viral entry into the brain following footpad inoculation with West Nile virus in mice. 1944 94


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