purification and characterization of a strain of coxsackievirus b4 of human origin that induces diabetes in mice.a diabetogenic strain of coxsackievirus b4 of human origin has been purified to study its biochemistry and diabetogenicity. tissue culture cells infected with the virus contain two distinct types of particles--virions and membrane-bound virions (mbv). mbvs are lighter (p = 1.29) than virions (p = 1.34), and they contain relatively more protein than rna. virons contain four capsid proteins, vpi-4, of various molecular weights: vp1, 37,500; vp2, 36,000; vp3, 26,000; and vp4, 5,500. mbvs contain th ...19882846770
membrane-bound virions of coxsackievirus b4: cellular localization, analysis of the genomic rna, genome-linked protein, and effect on host macromolecular synthesis.hela cells infected with several group b coxsackieviruses contain, in addition to standard virions, a population of virus-specific ribonucleoprotein particles which we (5) designated membrane-bound virions (mbv). mbvs differ from standard virions in buoyant density, yield, appearance, protein composition and infectivity. here we present several new features of mbvs of coxsackievirus b4. the mbvs are lighter (rho about 1.30) and are localized in rough membranes, intermixed with virions. they cont ...19852985028
protein kinase in nondiabetogenic coxsackievirus b4.alkali-dissociated, purified preparations of prototype coxsackievirus b4 release a protein kinase that catalyzes the incorporation of gamma-phosphate from 32p-labeled atp into three virus capsid proteins (vp1, vp3, vp4), several additional proteins of the particle, and exogenous acceptor proteins. using protamine sulfate as an acceptor protein, we detected nearly 20-fold more enzyme activity in membrane-bound virions (mbv) than in virions of the virus. the activity in the mbv is cyclic nucleotid ...19863018140
isolation and characterization of a membrane-bound population of group b coxsackieviruses.hela cells infected with several group b coxsackieviruses contain a previously undetected, virus-specific ribonucleoprotein particle which we designated membrane-bound virion (mbv). mbvs of b5 virus have a pronounced polygonal appearance and are slightly smaller than virions. the particles sediment more slowly (at about 107s) and have a lower buoyant density (about 1.30). they contain 35s virion rna; only three, and not four, capsid proteins; and at least seven additional proteins with apparent ...19836300437
pcr analysis of the viral complex associated with la france disease of agaricus bisporus.reverse transcription pcr analysis was used to investigate the involvement of two rna-genome viruses, la france isometric virus (liv) and mushroom bacilliform virus (mbv), in the etiology of la france disease of the cultivated mushroom agaricus bisporus. reverse transcription pcr amplification of sequences targeted to the genomes of liv and mbv, with a sensitivity of detection of < 10 fg of viral rna, showed diseased mushrooms to be either singly infected by liv or doubly infected by liv and mbv ...19957793952
use of non-radioactively labeled dna probes for the detection of a baculovirus from penaeus monodon by in situ hybridization on fixed tissue.clones isolated from a genomic library of the baculovirus pmsnpv (= mbv) were used to prepare dna probes for detection of pmsnpv in shrimp tissue. davidson's afa fixed shrimp tissues were probed using dna that had been labeled with the hapten, digoxigenin, by an in situ hybridization assay. the dna probes readily distinguished pmsnpv-infected from uninfected shrimp tissue. the reactions were localized to the hepatopancreas and midgut, the sites where this virus is detected by standard histologic ...19947822460
the nucleotide sequence and genome organization of mushroom bacilliform virus: a single-stranded rna virus of agaricus bisporus (lange) imbach.mushroom bacilliform virus (mbv) is found in association with spherical virus-like particles in cultivated mushrooms (agaricus bisporus) afflicted with la france disease. mbv possesses a monopartite ssrna genome of positive sense and differs from the majority of characterized mycoviruses, which contain segmented dsrna genomes. we have cloned and sequenced the mbv genome and determined that its length is 4009 nucleotides. the mbv genome contains four major and three minor open reading frames and ...19948030251
development of a non-radioactive gene probe by pcr for detection of white spot syndrome virus (wssv).combining primers created from the sequence information of two baculo-like viruses of penaeid shrimp, baculovirus penaei (bp) and monodon baculovirus (mbv), produced a 750 bp band on a 0.8% agarose gel using white spot syndrome virus (wssv), from penaeus monodon, as the dna template. the pcr fragment was ligated to a plasmid vector, (pgem-t) and transformed, creating a 3.7 kbp clone. the dna insert was sequenced, and the original primer pair was located. using restriction enzymes, the insert was ...19979015290
colourimetric pcr-based detection of monodon baculovirus in whole penaeus monodon postlarvae.the development of a nested polymerase chain reaction (pcr) assay is described to detect low concentrations of monodon baculovirus (mbv) dna from total penaeus monodon postlarval dna. a modified dna extraction procedure was also developed to circumvent problems associated with co-purification of pcr inhibitors in total dna extracted from whole postlarvae. this method involved mechanical disruption of frozen prawn material immediately followed by phenol extraction at high temperature. an assessme ...19989763125
mushroom bacilliform virus rna: the initiation of translation at the 5' end of the genome and identification of the vpg.mushroom bacilliform virus (mbv) is often found in cultivated mushrooms (agaricus bisporus) with la france disease. mbv has a 4-kb ssrna genome of positive-sense encoding four major open reading frames (orfs). the arrangement of orfs at the 5' end of the genome and the deduced amino-acid sequences of two of the putative gene products (protease and rna-dependent rna polymerase) show remarkable similarity to some plant viruses, particularly subgroup ii luteoviruses. we show that this similarity ex ...19989791015
identification of a subgenomic mrna encoding the capsid protein of mushroom bacilliform virus, a single-stranded rna mycovirus.mushroom bacilliform virus is unique among mycoviruses of the higher fungi in having a genome of positive-sense single-stranded rna. we have identified a subgenomic mrna molecule encoding the viral capsid protein in mushroom bacilliform virus-infected mycelium of agaricus bisporus. transcription of subgenomic rna commences at the sequence acaaaa, 47 nucleotides upstream of the initiating aug.199910417262
diagnosis of penaeus monodon-type baculovirus by pcr and by elisa of occlusion bodies.the black tiger prawn penaeus monodon is a valuable aquaculture product in taiwan. two specific diagnostic methods were established for p. monodon-type baculovirus, one using polymerase chain reaction (pcr) technology and the other enzyme-linked immunosorbent assay (elisa) technology. monodon-type baculovirus (mbv) was purified by sucrose gradient centrifugation from occlusion bodies of mbv-infected postlarvae of p. monodon. mbv dna was subsequently purified from the occlusion bodies and its pre ...200010782342
multiple viral infection in penaeus monodon shrimp postlarvae in an indian hatchery.moribund penaeus monodon postlarvae (pl8-pl10) in a hatchery in india were found to be simultaneously infected by 3 different viruses. they were highly infected with monodon baculovirus (mbv) and hepatopancreatic parvovirus (hpv) by histology and with white spot syndrome virus (wssv) by non-nested polymerase chain reaction (pcr). apparently healthy postlarvae tested from the same hatchery were positive for mbv and wssv by nested pcr only. tissue sections of such postlarvae did not show any histo ...200212033710
detection by pcr of hepatopancreatic parvovirus (hpv) and other viruses in hatchery-reared penaeus monodon postlarvae.the prevalence of hepatopancreatic parvovirus (hpv), monodon baculovirus (mbv) and white spot syndrome virus (wssv) in samples of penaeus monodon postlarvae (pl10 to pl20, 10 to 20 d old postlarvae) in india was studied by pcr. samples collected from different hatcheries, and also samples submitted by farmers from different coastal states, were analyzed. hpv was detected in 34%) of the hatchery samples and 31% of the samples submitted by farmers, using a primer set designed for detection of hpv ...200314735932
appendage deformity syndrome--a nutritional disease of macrobrachium rosenbergii.culture of the freshwater prawn macrobrachium rosenbergii as an alternative to penaeid shrimp has recently increased in coastal areas of southern india in order to avoid numerous problems, particularly with white spot syndrome virus (wssv). however, m. rosenbergii culture is now threatened by a new disease, appendage deformity syndrome (ads), that also results in high mortality. analysis of ads prawns for viruses such as wssv, monodon baculovirus (mbv) and infectious hypodermal and hematopoeitic ...200415212295
multiple pathogens found in growth-retarded black tiger shrimp penaeus monodon cultivated in 2001-2002 throughout thailand, black tiger shrimp penaeus monodon farmers reported very unusual retarded growth. we have called this problem monodon slow growth syndrome (msgs). based on decreased national production, estimated losses due to this phenomenon were in the range of 13 000 million baht (approximately 300 million us dollars) in 2002. since rearing practices had not changed, it was considered possible that the msgs problem may have arisen from a new or existing pathogen. to examine ...200415460852
development of real-time polymerase chain reaction assay with taqman probe for the quantitative detection of infectious hypodermal and hematopoietic necrosis virus from assay was developed for the detection of infectious hypodermal and hematopoietic necrosis virus (ihhnv) based on real-time quantitative polymerase chain reaction (pcr). a pair of primers and a taqman probe were designed that are specific for the recognition of a conservative region in the ihhnv genome. the ihhnv real-time pcr assay had a detection limit of 9 dna copies, with a dynamic range of detection between 9 x 106 and 9 dna copies. the primer pairs and probe were specific to ihhnv and di ...200616512254
detection of laem-singh virus (lsnv) in cultured penaeus monodon from india.viral diseases have become a major constraint on the growth of shrimp aquaculture in india. during a study of new diseases in cultured tiger shrimp penaeus monodon, diseased samples randomly collected from the southwestern and southeastern coasts of india were analyzed for white spot syndrome virus (wssv), monodon baculovirus (mbv), and hepatopancreatic parvovirus (hpv) by nested pcr, and for laem-singh virus (lsnv) by reverse transcription pcr (rt-pcr). of the 56 samples analyzed, 3 were positi ...200717933400
expression of the human cytomegalovirus ul97 gene in a chimeric guinea pig cytomegalovirus (gpcmv) results in viable virus with increased susceptibility to ganciclovir and lieu of a licensed vaccine, antivirals are being considered as an intervention to prevent congenital human cytomegalovirus (hcmv) infection. ideally, antiviral therapies should undergo pre-clinical evaluation in an animal model prior to human use. guinea pig cytomegalovirus (gpcmv) is the only small animal model for congenital cmv. however, gpcmv is not susceptible to the most commonly used hcmv antiviral, ganciclovir (gcv), rendering in vivo study of this agent problematic in the guinea pig ...200818325607
multiplex rt-pcr assay for simultaneous detection of six viruses of penaeid the present study, multiplex reverse transcription-polymerase chain reaction (mrt-pcr) was developed for simultaneously detection of six major shrimp viruses including yellow-head virus (yhv), white spot syndrome virus (wssv), taura syndrome virus (tsv), hepatopancreatic parvovirus (hpv), infectious hypodermal and hematopoietic necrosis virus (ihhnv) and monodon baculovirus (mbv). the six primer sets could amplify viral nucleic acids resulting in pcr products with different sizes. they were h ...200818406570
specific monoclonal antibodies raised against taura syndrome virus (tsv) capsid protein vp3 detect tsv in single and dual infections with white spot syndrome virus (wssv).the gene sequence encoding vp3 capsid protein of taura syndrome virus (tsv) was cloned into pgex-6p-1 expression vector and transformed into escherichia coli bl21. after induction, recombinant gst-vp3 (rvp3) fusion protein was obtained and further purified by electro-elution before use in immunizing swiss mice for production of monoclonal antibodies (mab). one mab specific to glutathione-s-transferase (gst) and 6 mab specific to vp3 were selected using dot blotting and western blotting. mab spec ...200818429444
purification and partial characterization of a bacilliform virus from agaricus bisporus: a single-stranded rna mycovirus.a bacilliform virus from the cultivated mushroom, agaricus bisporus, has been extensively purified and shown to be a unique mycovirus in that the genome is single-stranded (ss) rna. the purification procedure entailed extraction at ph 6, followed by polyethylene glycol-nacl precipitation, and differential, rate-zonal, and equilibrium centrifugation. mushroom bacilliform virus (mbv) has a buoyant density of 1.317 g/cm(3) in cs2so4, a260/a280 nm absorbance ratio of 1.67, and contains approximately ...198018631673
shrimp hepatopancreatic parvovirus detection by combining loop-mediated isothermal amplification with a lateral flow dipstick.present methods such as traditional pcr, pcr-elisa, real-time pcr and histopathology for detection of shrimp hepatopancreatic parvovirus (pmdnv) entail various disadvantages including high cost, long assay time or use of toxic substances. loop-mediated isothermal amplification (lamp) of target nucleotide sequences under inexpensive isothermal conditions combined with amplicon detection by chromatographic lateral flow dipsticks (lfd) allowed simpler detection within 75 min. biotinylated lamp ampl ...200818835299
diverse cytomegalovirus ul27 mutations adapt to loss of viral ul97 kinase activity under vitro resistance to maribavir (mbv), a cytomegalovirus ul97 kinase inhibitor currently in clinical trials, is known to result from viral ul97 mutations that confer moderate to high-level resistance and ul27 mutations that confer low-level resistance. to add to the four reported ul27 mutations, cytomegalovirus isolates or strains were propagated under mbv. four clinical isolates evolved ul27 mutations, which were first detected after 8 to 30 passages under drug selection. in three separate exp ...200918981262
detection of shrimp infectious myonecrosis virus by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick.infectious myonecrosis virus (imnv) has caused a slowly progressive disease with cumulative mortalities of up to 70% or more in cultured penaeus (litopenaeus) vannamei in northeast brazil and indonesia. rapid detection of viruses by loop-mediated isothermal amplification (lamp) of genomic material with high specificity and sensitivity can be applied for diagnosis, monitoring and control of diseases in shrimp aquaculture. using an imnv template, successful detection was achieved after a 60-min rt ...200919022295
development of a real-time pcr assay for detection of monodon baculovirus (mbv) in penaeid shrimp.a real-time pcr method was developed to detect monodon baculovirus (mbv) in penaeid shrimp. a pair of mbv primers to amplify a 135bp dna fragment and a taqman probe were developed. the primers and taqman probe were specific for mbv and did not cross react with hepatopancreatic parvovirus (hpv), white spot syndrome virus (wssv), infectious hypodermal and haematopoietic virus (ihhnv) and specific pathogen free (spf) shrimp dna. a plasmid (pmbv) containing the target mbv sequence was constructed an ...200919622362
rapid and sensitive detection of penaeus monodon nucleopolyhedrovirus by loop-mediated isothermal amplification.loop-mediated isothermal amplification (lamp) is a novel, sensitive and rapid method for amplification of nucleic acids under isothermal conditions. in this report, a lamp method was developed for detection of penaeus monodon nucleopolyhedrovirus (pemonpv), known previously as monodon baculovirus (mbv), using a set of six primers designed to specifically recognize the pemonpv polyhedrin gene. the optimized time and temperature conditions for the lamp assay were 60 min at 63 degrees c. the sensit ...200919703492
maribavir inhibits epstein-barr virus transcription in addition to viral dna replication.although many drugs inhibit the replication of epstein-barr virus (ebv) in cell culture systems, there is still no drug that is effective and approved for use in primary ebv infection. more recently, maribavir (mbv), an l-ribofuranoside benzimidazole, has been shown to be a potent and nontoxic inhibitor of ebv replication and to have a mode of action quite distinct from that of acyclic nucleoside analogs such as acyclovir (acv) that is based primarily on mbv's ability to block the phosphorylatio ...200919759127
rapid and sensitive detection of penaeus monodon nucleopolyhedrovirus (pemonpv) by loop-mediated isothermal amplification combined with a lateral-flow dipstick.several methods such as traditional pcr or nested-pcr, immuno assay and histopathology have been developed for detection of penaeus monodon nucleopolyhedrovirus (pemonpv) formerly called monodon baculovirus (mbv). however, these methods have various disadvantages including low sensitivity, long assay time, use of toxic substances or unsuitability for field diagnosis. loop-mediated isothermal amplification of target nucleotide sequences under isothermal conditions, combined with amplicon detectio ...201019818396
monodon baculovirus (mbv) infects the freshwater prawn macrobrachium rosenbergii cultivated in thailand.field specimens of post-larvae of the giant freshwater prawn (macrobrachium rosenbergii) from thailand showed hepatopancreatic tubule epithelial cells that contained central, eosinophilic inclusions within enlarged nuclei and marginated chromatin. these inclusions resembled those produced by some baculoviruses prior to formation of occlusion bodies that enclose virions in a polyhedrin protein matrix. by electron microscopy, the intranuclear inclusions contained bacilliform, enveloped virions (ap ...201019963025
fast short-fragment pcr for rapid and sensitive detection of shrimp viruses.this article describes a fast short-fragment pcr method for the detection of white spot syndrome virus (wssv), infectious hypodermal and hematopoietic necrosis virus (ihhnv), and monodon baculovirus (mbv). fast two-temperature (95 degrees c denaturation and 60 degrees c annealing/extension) pcrs were performed in 5-10 microl volume samples in miniaturized microplates using a fast peltier thermal cycler. 40 cycles were completed in 25-30 min. rapid high-resolution agarose gel electrophoresis of 7 ...201020471423
duplex real-time pcr for detection and quantification of monodon baculovirus (mbv) and hepatopancreatic parvovirus (hpv) in penaeus monodon.we describe a duplex real-time pcr assay using taqman probes for the simultaneous detection of monodon baculovirus (mbv) and hepatopancreatic parvovirus (hpv). both mbv and hpv are shrimp enteric viruses that infect intestinal and hepatopancreatic epithelial cells. both viruses can cause significant mortalities and depressed growth in infected larval, postlarval, and early juvenile stages of shrimp, and thus present a risk to commercial aquaculture. in this duplex assay, we combined 2 single rea ...201121516971
progress in the development of new therapies for herpesvirus infections.resurgent interest in antiviral drugs for the treatment of herpesvirus has led to the development of new compounds that are progressing through clinical trials. this is important because there are few therapeutic options for resistant infections and some viruses such as human cytomegalovirus remain underserved. new compounds include conventional dna polymerase inhibitors such as valomaciclovir and cyclopropavir, as well as cmx001 that has a broad spectrum of antiviral activity that includes all ...201122162744
Impact of mineral deposition on shrimp, Penaeus monodon in a high alkaline water.This study compares water quality parameters, shrimp growth and mortality rates, and biomass at harvest in two ponds of equal size, seeded with the same density (7 m2) of White Spot Syndrome Virus (WSSV) and Monodon Baculo Virus (MBV) negative post-larvae (PL)-20 of shrimp, Penaeus monodon in the Vellar estuary of South India. The primary difference between the ponds was the water source; one was filled from the estuary and the second with water from bore wells with high alkalinity. Temperature ...201122167938
the genome of oryctes rhinoceros nudivirus provides novel insight into the evolution of nuclear arthropod-specific large circular double-stranded dna viruses.the oryctes rhinoceros nudivirus (ornv) is a dsdna virus with enveloped, rod-shaped virions. its genome is 127,615-ábp in size and contains 139 predicted protein-coding open reading frames (orfs). in-depth genome sequence comparisons revealed a varying number of shared gene homologues, not only with other nudiviruses (nvs) and baculoviruses, but also with other arthropod-specific large dsdna viruses, including the so-called monodon baculovirus (mbv), the salivary gland hypertrophy viruses (sghvs ...201121380757
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