Abstract：
:Single-stranded conformation polymorphism (SSCP) is a technique used widely for the detection of differences in DNA sequence based on PCR technology. Developed by geneticists for the detection of mutations causing disease, it has been adopted more recently for the analysis of the quasi-species of viral genomes, such as hepatitis C virus (HCV). The rigorous standardisation and determination of the limit of detection of the technique has rarely been shown. Variants within the quasi-species of the hypervariable region of HCV were cloned into pUC119 and the resulting plasmids quantitated and used as templates to optimise SSCP. Variables studied included the number of variants detected, the sensitivity of detection of variants in the minority, the electrophoresis temperature, methods of generation of single-stranded DNA, effect of numbers of cycles of PCR and use of DNA polymerase with proof-reading ability. It was demonstrated that the optimised method could detect at least five variants within a quasi-species and that variants could be detected down to a level of 2% of the quasi-species. Electrophoresis at room temperature for 18 h was highly reproducible. Generation of single-stranded DNA using a single primer with Taq polymerase for 20 cycles gave an accurate reflection of the quasi-species make-up and use of Pfu polymerase reduced sensitivity of detection of minor bands. This SSCP method provides an accurate tool to evaluate HCV quasi-species.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
McKechnie VM,McCruden EAdoi
10.1016/s0166-0934(00)00279-2subject
Has Abstractpub_date
2001-04-01 00:00:00pages
131-9issue
2eissn
0166-0934issn
1879-0984pii
S0166093400002792journal_volume
92pub_type
杂志文章abstract：:Formalinized chick erythrocytes agglutinated with Japanese encephalitis virus antigens over a wider range of pH (6.0-7.2) than fresh erythrocytes (pH 6.4-6.8). The virus antigens were unstable below pH 7.0 and therefore the use of the formalinized erythrocytes made it possible for a one-diluent system of the hemagglut...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(80)90026-9
更新日期：1980-01-01 00:00:00
abstract：:The Directigen Flu-A is an enzyme immunoassay for detecting in 15 min the influenza A nucleoproteinic antigen directly from specimens after passive adsorption on a cellulose membrane. The test was assessed using 160 frozen (-20 degrees C) specimens collected during the 1988-1989 A/H1N1 influenza epidemic and the 1989-...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(92)90033-a
更新日期：1992-06-01 00:00:00
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journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(82)90012-x
更新日期：1982-11-01 00:00:00
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journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2013.04.001
更新日期：2013-07-01 00:00:00
abstract：:Fowl adenovirus serotype-4 (FAdV-4) has been recognized as a predominant threat to the broilers aged from three to five weeks. Hydropericardium syndrome (HPS) is one of its major clinical diseases by FAdV-4 resulting in heavy economic losses. In this study, a loop-mediated isothermal amplification coupling with a late...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2019.04.026
更新日期：2019-08-01 00:00:00
abstract：:The recent emergence of novel viruses requires reliable methodology for their identification and confirmation both on a cellular and molecular level. Mass spectrometry offers a suitable approach for the identification and characterisation of viral proteins and its application is demonstrated in this study. Menangle vi...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(01)00441-4
更新日期：2002-04-01 00:00:00
abstract：:We show that the immuno-gold technique adapted to electron microscopy is a sensitive method for in situ localization of viral proteins in plant cells. Using antisera raised against cauliflower mosaic virus and the viroplasm major protein (VmP) we obtained a selective labelling of the viral particles and of the viropla...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(84)90053-3
更新日期：1984-12-01 00:00:00
abstract：:Human enteroviruses are among the most common viruses infecting humans. These viruses are known to be able to infect a wide range of tissues and are believed to establish persistent infections. Enteroviruses are positive-sense single-stranded RNA viruses whose replication involves the synthesis of negative strand inte...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2008.07.010
更新日期：2008-11-01 00:00:00
abstract：:Cutting edge immune monitoring techniques increasingly measure multiple functional outputs for various cell types, such as intracellular cytokine staining (ICS) assays that measure cytokines expressed by T cells. To date, however, there is no precise method to measure virus-specific cytokine production by both T cells...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.01.003
更新日期：2014-04-01 00:00:00
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journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2018.01.004
更新日期：2018-04-01 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 leukemia virus (BLV) is widely distributed in U.S. cattle herds. It infects B lymphocytes and causes neoplastic disease in 5-10% of infected animals. Direct economic losses are incurred as a result of death, reduced milk production and condemnation at slaughter. Thus the identification of cattle infected with B...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(91)90009-o
更新日期：1991-06-01 00:00:00
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journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2021.114062
更新日期：2021-01-08 00:00:00
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journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2010.07.040
更新日期：2010-10-01 00:00:00
abstract：:The World Health Organization recommends screening donor blood for HIV in centralized laboratories. This recommendation contributes to quality, but presents specimen transport challenges for resource-limited settings which may be relieved by using dried blood spots (DBS). In sub-Saharan Africa, most countries screen d...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2011.07.001
更新日期：2012-01-01 00:00:00
abstract：:The simple spot hybridization technique to detect HBV DNA in serum was modified to concentrate Dane particles by pelleting. This minimised interference by serum components and allowed larger volumes of sera (up to 500 microliters) to be used in serial dilution on borderline positive samples and increased the efficienc...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(85)90136-3
更新日期：1985-12-01 00:00:00
abstract：:Nine commercial EIAs for measles-specific IgG were compared with haemagglutination inhibition (HI) and plaque reduction neutralization (PRN). A total of 174 sera selected, to give approximately half of the sera without measles antibody by HI, were tested by all EIAs and HI. However, there was sufficient volume of only...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(97)02210-6
更新日期：1997-06-01 00:00:00
abstract：:Water is one major pathways by which the white spot syndrome virus (WSSV) pathogen enters aquaculture facilities. This paper describes the production and use of a capacitive biosensor for the quantitative detection of as little as 1copy/μl of WSSV in shrimp pond water. A glutathione-S-transferase tag for white spot bi...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2011.01.010
更新日期：2011-04-01 00:00:00
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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：:Portable user-friendly diagnostic tests can benefit detection and surveillance of wildlife diseases. Here, the performance of a compact POCKIT™ Nucleic Acid Analyzer for detection of Ambystoma tigrinum virus (ATV), an emerging Iridovirus that is associated with high host mortality in the western tiger salamander (Amby...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2017.08.008
更新日期：2017-11-01 00:00:00
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journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2010.11.005
更新日期：2011-01-01 00:00:00
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journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2011.02.015
更新日期：2011-05-01 00:00:00
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journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(96)02165-9
更新日期：1997-04-01 00:00:00
abstract：:Understanding the ability to survive in an aerosol leads to better understanding of the hazard posed by pathogenic organisms and can inform decisions related to the control and management of disease outbreaks. This basic survival information is sometimes lacking for high priority select agents such as the filoviruses ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2011.06.021
更新日期：2011-10-01 00:00:00
abstract：:A new procedure for the PCR amplification of unknown DNA sequences adjacent to a known sequence is described. The required but not readily available second primer sequence in the unknown DNA sequence is obtained by creating an overhanging restriction site in the unknown sequence to which a double-stranded oligonucleot...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(91)90191-2
更新日期：1991-04-01 00:00:00
abstract：:We have developed a sensitive method to detect and localize HPV-6 viral DNA, mRNA and protein in biopsy specimens of genital and respiratory tract lesions by using in situ hybridization and immunoperoxidase assays on sections of plastic-embedded tissue. This modified in situ hybridization technique, using ultrathin se...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(89)90098-0
更新日期：1989-07-01 00:00:00
abstract：:A polymerase chain reaction (PCR) based strategy to detect episomal banana streak badnavirus (BSV) in banana and plantain plants that carry integrated BSV sequences was developed. Antisera used in immuno-capture polymerase chain reaction (IC-PCR) are capable of binding a large number of BSV serotypes. The primers used...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(98)00158-x
更新日期：1999-04-01 00:00:00
abstract：:A stably transformed cell line (BHKICP6LucA6) has been isolated which expresses high levels of luciferase activity following infection with herpes simplex virus (HSV). The genome of this cell line contains an HSV-1 promoter-luciferase chimeric gene. Infected BHKICP6LucA6 cells exhibit a level of luciferase activity 5 ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(94)90071-x
更新日期：1994-04-01 00:00:00
abstract：:A recombinant hepatitis B surface antigen (HBsAg) has been produced in the yeast Saccharomyces cerevisiae and used as a vaccine against hepatitis B virus (HBV) infection. The present study aimed to optimize the extraction of recombinant virus-like particles (rVLPs) to develop a simple purification procedure based on g...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2012.09.023
更新日期：2013-01-01 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
