Abstract:
:HIV-1 RNA viral load is the preferred tool to monitor virological failure during antiretroviral therapy (ART) exposure. Timely detection of virological failure can reduce the prevalence and complexity of HIV-1 drug resistance. This field evaluation further characterizes a two-step approach to identify virological failure, as a measure of ART adherence, and detect HIVDR mutations in the reverse transcriptase (RT) gene of HIV-1. Two hundred and forty-eight (248) samples were tested; 225 from South African HIV-1 participants enrolled in the PharmAccess African Studies to Evaluate Resistance (PASER) cohort, forty of which had paired dried blood spot (DBS) samples and 23 HIV-1 negative samples. A newly developed virological failure assay (ARTA-VFA) was used on all samples, and those with a viral load >5000 RNA copies/ml were genotyped with a shortened RT protocol to detect HIVDR (ARTA-HIVDR(ultralight)). The ARTA-VFA showed good precision and linearity as compared to a commercial reference assay (NucliSENS EasyQ v1.2, Roche) with an R(2) of 0.99. Accuracy studies illustrated standard deviations of <1 log RNA copies/ml for plasma and DBS ARTA-VFA results compared to the reference method. The ARTA-VFA's intended use was to deliver qualitative results either < or >5000 RNA copies/ml. No significant differences in the proportion of results < or > either the 5000 RNA copies/ml or 1000 RNA copies/ml cut-off were noted for plasma indicating either cut-off to be useful. Significant differences were noted in these proportions when DBS were used (P=0.0002), where a 5000 RNA copies/ml cut-off was deemed more appropriate. The sensitivity and specificity of the ARTA-VFA with plasma were 95% and 93% and 91% and 95% for DBS using a 5000 RNA copies/ml cut-off. The ARTA HIVDR(ultralight) assay was reliable for plasma and DBS samples with a viral load >5000 RNA copies/ml, with amplification and sequencing success rates of 91% and 92% respectively for plasma, and 95% and 80% respectively for DBS. HIVDR profiles for plasma and DBS were 100% concordant with the reference assay. This study evaluated a previously described combination of two assays potentially useful in assessing HIV-1 virological failure and resistance, showing good concordance with reference assays. These assays are simple to perform and are affordable, viable options to detect virological failures in certain resource limited settings. The assays' compatibility with DBS sampling extends the access of HIV-1 virological monitoring to more remote settings.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Bronze M,Aitken SC,Wallis CL,Steegen K,Stuyver LJ,de Wit TF,Stevens Wdoi
10.1016/j.jviromet.2013.08.015subject
Has Abstractpub_date
2013-12-01 00:00:00pages
300-7issue
1-2eissn
0166-0934issn
1879-0984pii
S0166-0934(13)00347-9journal_volume
194pub_type
杂志文章abstract::Accurate detection and quantitation of viruses can be beneficial to plant-virus interaction studies. In this study, three SYBR green real-time RT-PCR assays were developed to quantitate grapevine leafroll-associated virus 3 (GLRaV-3) in infected vines. Three genomic regions (ORF1a, coat protein and 3'UTR) were targete...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2014.09.022
更新日期:2014-12-15 00:00:00
abstract::Tomato yellow leaf curl Sardinia virus (TYLCSV) (Geminiviridae) is an important pathogen severely affecting tomato production in the Mediterranean basin. Although diagnostic protocols are available for its detection in plants and its vector Bemisia tabaci (Gennadius), suitable tools for estimating and comparing viral ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2007.09.015
更新日期:2008-02-01 00:00:00
abstract::Canine serum preserved at room temperature (25°C) for longer than 24h is known to exhibit significant cytotoxicity. This phenomenon is one of the major reasons for the failure of virus neutralization tests. In this study, a method for reducing this cytotoxicity was investigated by applying several treatments to dog, c...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2015.08.018
更新日期:2015-12-01 00:00:00
abstract::Infectious bursal disease virus (IBDV) is an immunosuppressive disease of young chicken characterized by severe depletion of B-lymphocytes in the bursa of Fabricius. To provide antigen for diagnostic tests, its major structural protein VP2 was expressed in the yeast Saccharomyces cerevisiae. Electron microscopy of pur...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2008.11.020
更新日期:2009-04-01 00:00:00
abstract::A recombinant murine cytomegalovirus (mCMV) that expresses enhanced green fluorescent protein (EGFP) under control of the native immediate-early 1/3 promoter was constructed to detect directly sites of viral activity in latent and reactivated infections. The recombinant virus had acute and latent infection characteris...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(00)00202-0
更新日期:2000-09-01 00:00:00
abstract::Chronic infection of the hepatitis B virus (HBV) is one of the causes leading to liver cancer. The 3.2kb genome of HBV encodes four proteins: core antigen, surface antigen, a DNA polymerase and the X protein (HBx). The biological functions of HBx are not fully understood. It has been shown that HBx is a potent trans-a...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2005.01.022
更新日期:2005-06-01 00:00:00
abstract::HTLV-1 and HTLV-2 are highly related delta-retroviruses that infect and transform T-lymphocytes, but have distinct pathogenic properties. HTLV replication and survival requires the expression of multiple gene products from an unspliced and a series of highly related alternatively spliced mRNA species. To date, the com...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2007.01.023
更新日期:2007-06-01 00:00:00
abstract::A real-time TaqMan RT-PCR assay was developed for the rapid and sensitive detection of Tomato ringspot virus (ToRSV), an important plant virus which infects a wide range of fruit and ornamental crops. Primers and a probe were designed based on the highly conserved 3'-untranslated region (UTR) sequences of ToRSV, to am...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2014.02.011
更新日期:2014-06-01 00:00:00
abstract::To develop reverse genetics system of RNA viruses, cloning of full-length viral genome is required which is often challenging due to many steps involved. In this study, we report cloning of full-length cDNA from an Indian field isolate (CSFV/IVRI/VB-131) of classical swine fever virus (CSFV) using in vitro overlap ext...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2015.10.006
更新日期:2015-12-15 00:00:00
abstract::The polymerase chain reaction (PCR) methods enable the detection of large number of human papillomavirus (HPV) genotypes that infect the anogenital tract. In this study, two groups of cervical scrapes with abnormal cytomorphology were analysed. The first group was tested with three sets of consensus primers located wi...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(00)00194-4
更新日期:2000-08-01 00:00:00
abstract::The aim of this study was to evaluate viral DNA load in paediatric patients with herpes simplex infections of the central nervous system. A real-time PCR assay for herpes simplex types 1 and 2 was developed for this 8-year retrospective study that included children with herpes simplex infection of the central nervous ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2007.09.009
更新日期:2008-02-01 00:00:00
abstract::A real-time PCR assay, which enables simultaneous detection and differentiation of all three serotypes of Marek's disease virus, without the need for post-PCR sequencing, has been developed. The assay is based on the primer-probe energy transfer real-time PCR, which has a relatively high tolerance towards point mutati...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2009.12.012
更新日期:2010-04-01 00:00:00
abstract::Canine distemper, caused by Canine distemper virus (CDV), is a highly contagious and fatal systemic disease in free-living and captive carnivores worldwide. Accurate, rapid and simple detection of CDV is critical to improve disease management and prevent outbreaks. In this study, a visible and incubation instrument-fr...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2018.07.007
更新日期:2018-10-01 00:00:00
abstract::To map the epitopes of VP2 protein of chicken anemia virus (CAV), VP2 was expressed as a fusion protein in Escherichia coli BL21 (DE3). The Western blot demonstrated that recombinant VP2 protein could be recognized by sera of chickens infected with CAV. Female BALB/c mice were immunized with purified recombinant VP2 p...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2007.03.016
更新日期:2007-08-01 00:00:00
abstract::We have cloned and sequenced the glycoprotein B genes from five strains of BaCMV, isolated from three subspecies of cynocephalus baboons (olive, yellow and chacma). Primers were designed using conserved DNA regions of the gB gene to allow DNA amplification from all strains of BaCMV. These regions differ sufficiently f...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2005.01.010
更新日期:2005-05-01 00:00:00
abstract::Emergence of lamivudine (LAM) resistance causes treatment failure in patients with chronic hepatitis B and compromise the efficacy of subsequent salvage therapies with other nucleot(s)ide analogs (NAs). Establishment of cell-based assays supporting LAM-resistant hepatitis B virus (HBV) replication will not only provid...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2014.02.008
更新日期:2014-06-01 00:00:00
abstract::A real-time reverse transcription-polymerase chain reaction (qRT-PCR) assay was developed for the fast and highly sensitive detection of the sacbrood virus (SBV) genome and applied to honeybee samples. Using plasmid DNA containing a partial SBV genome and diluted serially, as few as 1×10(2)copies/μl (correlation co-ef...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2011.10.014
更新日期:2012-01-01 00:00:00
abstract::The plaque assay is essential for virion quantitation but the classic protocol requires considerable efforts. A simplified dengue 96-well plaque assay with automated quantitation program is an alternative to access the level of infectious virus. Dengue plaque assay was simplified using LLC/MK2 cells and virus mixing s...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2016.08.009
更新日期:2016-11-01 00:00:00
abstract::Addition of exogenous peptide sequences on viral capsids is a powerful approach to study the process of viral infection or to retarget viruses toward defined cell types. Until recently, it was not possible to manipulate the genome of mammalian reovirus and this was an obstacle to the addition of exogenous sequence tag...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2011.11.021
更新日期:2012-02-01 00:00:00
abstract::Henipaviruses were first discovered in the 1990s, and their potential threat to public health is of increasing concern with increasing knowledge. Old-world fruit bats are the reservoir hosts for these viruses, and spill-over events cause lethal infections in a wide range of mammalian species, including humans. In anti...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2009.05.014
更新日期:2009-10-01 00:00:00
abstract::Loop-mediated isothermal amplification is a novel method for rapid amplification of DNA. It has been adopted widely for the detection of virus because of its simplicity, rapidity, and specificity. A loop-mediated isothermal amplification assay was developed for the detection of porcine parvovirus. Four primers specifi...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2008.10.004
更新日期:2009-02-01 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::A method for quantitative analysis of the growth properties of human cytomegalovirus (HCMV) in various cell culture systems was developed. Recent HCMV isolates are, in most cases cell associated, causing only limited cytopathic effect. This renders comparative analysis of the biological properties of such isolates dif...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(97)02082-x
更新日期:1997-01-01 00:00:00
abstract::A sensitive enzyme-linked immunosorbent assay (ELISA) has been developed which can detect serotype and group-specific antibodies (IgG) to fowl adenovirus serotypes 2, 3 and 4. The chickens produced principally type-specific antibodies after a single oral inoculation of virus which enabled that strain to be identified ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(83)90086-1
更新日期:1983-12-01 00:00:00
abstract::Lily symptomless virus (LSV) occurs frequently in many Lilium species worldwide and often causes developmental abnormalities such as a smaller flower and lower bulb yield. In this study, two moderate and efficient gel filtration chromatography (GFC) methods was compared, these two techniques were, respectively, based ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2006.09.008
更新日期:2007-02-01 00:00:00
abstract::A real-time RT-PCR assay using SYBR Green was developed for specific and reliable quantitative detection of Citrus tristeza virus (CTV) in infected plants. A general primer set designed from conserved sequences in ORFs 1b and 2 enabled amplification of the genomic RNA (gRNA) while excluding most subgenomic and defecti...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2007.05.011
更新日期:2007-11-01 00:00:00
abstract::Human cytomegalovirus and human herpesvirus-6 are closely related viruses which cause similar diseases, have similar cellular repositories of latent infection, and may be detected largely in the same types of clinical specimens. DNA amplification appears likely to play an increasing role in the diagnosis of recent and...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(95)00019-q
更新日期:1995-06-01 00:00:00
abstract::One first generation assay (manufactured by Ortho, test I) and 3 second generation anti-HCV ELISAs (manufactured by Ortho, Abbott, and UBI, tests II-IV) were compared. Sera from 4 different sources were used: (1) intravenous drug-users (IVDUs, n = 50), (2) blood donors (n = 1055), (3) all clinical samples from one day...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(92)90087-t
更新日期:1992-12-01 00:00:00
abstract::Rapid detection of CMV-DNA in urine specimens by dot-blot hybridization was compared to conventional virus isolation and to virus identification using solid-phase immunoelectron microscopy (SPIEM). To detect viral DNA, 32P-labeled EcoR1 J fragment of CMV-DNA was used as a probe in the hybridization assay. In addition,...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(87)90028-0
更新日期:1987-05-01 00:00:00
abstract::The technique of gold-labelled immunosorbent electron microscopy for the initial screening of monoclonal antibodies 10 days after cell fusion from 96-well culture plates is described. The technique is used to identify clones that secrete antibodies binding on the surface of the virion or to viral subunits, and compare...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(88)90118-8
更新日期:1988-12-01 00:00:00