Abstract：
:Procedures were developed for the screening and isolation of RNA viruses that vary from the consensus population by a single point mutation and are present in low abundance. We tested these procedures using a mixture of the vaccine donor line cold-adapted (ca) B/Ann Arbor/1/66 and its wild type (wt) progenitor at a ratio of 1,000:1. A 13-mer oligodeoxynucleotide was prepared as a [32P]-radiolabeled probe which specifically hybridized to wt viral RNA (vRNA) at a region that varied between the ca and wt sequence by a single base at position 1,320 of the PA gene. This probe was able to detect as little as 88 pg of total wt vRNA blotted to nitrocellulose, while giving no positive signal with as much as 1 microgram of total ca vRNA. We were able to isolate virus containing the wt PA gene sequence from the mixed pool of ca and wt viruses by two successive rounds of amplification in embryonated eggs inoculated with a controlled number of infectious virions. During each round of amplification the abundance of the virus containing the wt PA gene sequence was increased about ten-fold. Once a relative abundance of 1:10 was reached, the virus was cloned by plaquing in Madin-Darby canine kidney cells. These procedures, which allow the isolation of specific point mutants, utilize no selective pressure conditions and are suitable for analyzing the phenotypic importance of known mutations in biologically important viruses.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Mitchell BM,DeBorde DCdoi
10.1016/0166-0934(93)90031-lsubject
Has Abstractpub_date
1993-05-01 00:00:00pages
181-91issue
2-3eissn
0166-0934issn
1879-0984pii
0166-0934(93)90031-Ljournal_volume
42pub_type
杂志文章abstract：:Titrating infective virus is one of the most important and common techniques in virology. However, after many years of widespread use, the parameters governing the accuracy of titration values are still not well understood. It was found that under conditions currently used for virus titration, only a small percentage ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(90)90047-j
更新日期：1990-10-01 00:00:00
abstract：:Purified hepatitis B surface antigen (HBsAg) of subtype ay was solubilized in guanidinium chloride and submitted to chromatography on Sepharose 4B in the presence of guanidinium chloride. The polypeptides P1 (Mr = 24,000) and P2 (Mr = 29,000) were eluted in the same fraction with a minor contaminant (Mr = 40,000). Lar...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(82)90039-8
更新日期：1982-01-01 00:00:00
abstract：:Conditions for using slot-blot nucleic acid hybridization to quantitatively detect dengue-2 virus using a radiolabelled cDNA probe, pVV17, were determined. As little as 11 plaque-forming units of virus were detected using a hybridization mixture without formamide and performing the test at 70 degrees C. While predomin...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(87)90097-8
更新日期：1987-02-01 00:00:00
abstract：:The clinical sensitivity of the current anti-HIV assays is based for an important part on their reactivity with seroconversion panels. The most sensitive assay closes the seroconversion window as much as possible, thereby reducing the risk of transmitting false negative donations obtained from individuals infected rec...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(98)00126-8
更新日期：1998-12-01 00:00:00
abstract：:Group A rotaviruses (RVs) are important pathogens that cause acute, dehydrating gastroenteritis in infants and young children. In this study, a multiplex real-time polymerase chain reaction protocol using primers and TaqMan(®) probes specific for viral VP4 and VP7 genes was evaluated. This assay offers simultaneous ge...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2011.12.009
更新日期：2012-03-01 00:00:00
abstract：:This report describes the first time entire viral capsids have been purified using fast protein liquid chromatography (FPLC) techniques. The FPLC is used here to separate the two electrophoretic forms of cowpea mosaic virus. The capsid forms are shown to be separated by the Mono-Q column without damaging the capsids. ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(87)90011-5
更新日期：1987-07-01 00:00:00
abstract：:Prion disorders occur when endogenous prion protein (PrP(C)) undergoes a conformational change from a predominantly α-helix-rich structure to an insoluble β-sheet-rich structure (PrP(Sc)). The resulting PrP(Sc) then in some way facilitates the progressive transformation of nearby PrP(C) to PrP(Sc). In time this result...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2010.06.016
更新日期：2010-11-01 00:00:00
abstract：:Xenotropic murine leukemia virus (X-MuLV) is often used in retrovirus elimination studies involving rodent cells. Currently, X-MuLV is measured using a focus-forming assay on mink (MiCl1 S+L-) or cat (PG-4 S+L-) cell lines. An easier and quicker PG-4 cell plaque assay, which retains the statistical reproducibility of ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(99)00064-6
更新日期：1999-08-01 00:00:00
abstract：:Noroviruses are the leading cause of nonbacterial gastroenteritis outbreaks in the United States, some of which are caused by the ingestion of contaminated water. Detection and genotypic characterization of noroviruses is commonly performed by reverse transcription-polymerase chain reaction (RT-PCR) followed by sequen...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2008.03.010
更新日期：2009-03-01 00:00:00
abstract：:A RT-nested PCR that amplifies part of the conserved nucleoprotein gene of avian Paramyxovirus type 1 is described. The technique allowed the detection of pigeon Paramyxovirus type 1 (pPMV-1) virus directly from a wide range of infected chicken and pigeon organs, and should be able to detect typical Newcastle disease ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(02)00148-9
更新日期：2002-12-01 00:00:00
abstract：:In many countries phytosanitary regulations apply to Potato spindle tuber viroid, because it can cause serious diseases in potato and tomato crops. Other pospiviroids, some of which are distributed widely in ornamental crops, can cause similar diseases. Consequently, there is a need for a reliable and cost-effective g...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2012.09.004
更新日期：2013-01-01 00:00:00
abstract：:The applicability of the enzyme immunoassay technique to the detection of group and interspecies determinants of C-type retroviral proteins was tested. For this purpose four groups of C-type retroviruses (MuLV, FeLV, SiSV/GaLV, and BaEV/RD114) were assayed for group-specific and interspecies reactivities of their p30 ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(82)90008-8
更新日期：1982-11-01 00:00:00
abstract：:Hepatitis A virus (HAV) harvested from infected MRC-5 cells can hemagglutinate various species of erythrocytes at acid pH (Eckels et al., 1989). Further studies revealed that the majority of the hemagglutinin (HA) in MRC-5 and BS-C-1 cells was cell-associated. A simplified procedure for preparing HAV-HA consisted of c...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(90)90123-w
更新日期：1990-06-01 00:00:00
abstract：:Influenza reverse genetics plays vital roles in understanding influenza molecular characteristics and vaccine development. However, current influenza reverse genetics heavily depends on restriction enzyme and ligation for gene cloning. The traditional cloning process of influenza eight fragments for virus rescuing gen...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2015.06.001
更新日期：2015-09-15 00:00:00
abstract：:A detailed growth and purification scheme suitable for producing relatively large quantities of fully active, pure SV40 is presented together with data on recovery and purity at each step of the procedure. The scheme was designed to prevent the initial binding of virus to cell components as well as contamination of th...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(81)90051-3
更新日期：1981-10-01 00:00:00
abstract：:We assessed the performance of Abbott RealTime CMV assay (ARC) compared to Roche Cobas Amplicor CMV Monitor Test (RCM) for quantification of CMV in plasma of transplant patients. Commercial panels were used to test linearity, precision and interference and 83 clinical samples were used for the accuracy and precision a...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2015.08.010
更新日期：2015-12-01 00:00:00
abstract：:In the context of eradication of poliomyelitis the World Health Organization stimulates the development of inactivated polio vaccines based on attenuated virus strains. In addition to vaccine development, tests have to be designed to assess the vaccine quality. An important test is the identification test for poliovir...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2013.01.026
更新日期：2013-04-01 00:00:00
abstract：:The geographical distribution of human immunodeficiency virus type 1 (HIV-1) subtypes show, with the exception of some African Countries, a specific pattern. However, the significant phenomenon of migration to Western Countries, coupled to inter-ethnic blending, may result in a constant introduction and spread of nove...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2004.11.014
更新日期：2005-03-01 00:00:00
abstract：:Three tests were compared to diagnose herpes virus infection: electron microscopy (EM), viral culture (VC) and cytodiagnosis (Tzanck smear). The study comprised 67 patients with skin or mucous membrane lesions suggestive of herpes simplex virus (HSV) infection. The sensitivity of EM increased 25% after virus concentra...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(91)90106-a
更新日期：1991-10-01 00:00:00
abstract：:Six single-chain fragment variable (scFv) antibodies against infectious haematopoietic necrosis virus (IHNV) were selected from an antibody phage display library by phage display technology. The soluble scFv antibodies showed a molecular weight 32kDa by Western blot. Dot blot analysis revealed that the six scFv antibo...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.04.008
更新日期：2014-09-01 00:00:00
abstract：:A newly developed integrated cell culture reverse transcriptase quantitative PCR (ICC-RTqPCR) method and its applicability in UV disinfection studies is described. This method utilizes a cell culture system coupled with four RTqPCR assays to detect four species of human enterovirus (e.g., Enterovirus A, Enterovirus B,...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2018.05.008
更新日期：2018-08-01 00:00:00
abstract：:The development of a rapid and sensitive system for detecting influenza viruses is a high priority for controlling future epidemics and pandemics. Quantitative real-time PCR is often used for detecting various kinds of viruses; however, it requires more than 2h per run. Detection assays were performed with super high-...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2011.08.015
更新日期：2011-12-01 00:00:00
abstract：:Rabbits and guinea pigs were immunized with purified bovine rotavirus. Immunoglobulin G fractions of the resulting antisera were used in a standard four-layer solid-phase enzyme immunoassay (EIA) for rotavirus antigen in human faecal specimens. Samples negative for rotavirus in electron microscopy, when diluted in sta...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(82)90096-9
更新日期：1982-09-01 00:00:00
abstract：:The introduction of molecular diagnostic methods for investigation of gastroenteritis has significantly reduced the diagnostic gap. However, approximately 25% of cases of gastroenteritis remain undiagnosed even after screening for bacteria, parasites and viruses using the most sensitive PCR and RT-PCR methods availabl...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2009.01.006
更新日期：2009-06-01 00:00:00
abstract：:Studies were conducted to determine the performance of four dyes in assessing antiviral activities of compounds against three RNA viruses with differing cytopathogenic properties. Dyes included alamarBlue® measured by absorbance (ALB-A) and fluorescence (ALB-F), neutral red (NR), Viral ToxGlo™ (VTG), and WST-1. Viruse...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2017.03.012
更新日期：2017-08-01 00:00:00
abstract：:An efficient method for isolation of virus mutants with antigenically altered proteins is described. The method is based on the separation of viruses with wild-type and antigenically altered proteins by affinity chromatography using monoclonal antibodies (MAbs). A nonessential glycoprotein E (gE) of Aujeszky's disease...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(98)00137-2
更新日期：1999-01-01 00:00:00
abstract：:A genetically modified cell line (Vero-ICP10-EGFP) was constructed for detection of herpes simplex virus (HSV) by a simple, rapid and direct method. The cell line was developed by stable transfection of Vero cell with a plasmid encoding the green fluorescent protein (GFP) driven by the promoter of the HSV-2 ICP10 gene...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(00)00234-2
更新日期：2000-11-01 00:00:00
abstract：:Monoclonal antibodies (Mabs) against influenza B virus were obtained by immunizing mice with B/Nagasaki/1/87, one of the strains of the B/Victoria group. Immunoprecipitation analysis revealed that individual Mabs precipitated the nucleoprotein (NP), the matrix protein (M) or the hemagglutinin protein (HA). By using th...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(99)00015-4
更新日期：1999-04-01 00:00:00
abstract：:Bovine viral diarrhea virus (BVDV) can contaminate biological products produced in bovine or porcine cells or manufactured using bovine sera. A rapid, specific, sensitive, and practical method of detecting BVDV in bio-products is needed. The purpose of this study was to compare three assays with respect to their abili...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.05.020
更新日期：2014-10-01 00:00:00
abstract：:Real-time PCR assays for nucleic acid testing (NAT) of hepatitis viruses A-E and for HIV-1 and HIV-2 have been developed; however, a multiplex assay that can simultaneously detect all of these agents is not yet available. Standardized TaqMan assays for detection of hepatitis viruses A-E have been described and applied...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2018.06.001
更新日期：2018-09-01 00:00:00
