Cost-effective HIV-1 virological monitoring in resource-limited settings using a modified commercially available qPCR RNA assay.

Abstract:

:Virological monitoring through plasma viral load (PVL) quantification is essential for clinical management of HIV patients undergoing antiretroviral treatment (ART), and for detecting treatment failure. Quantitative PCR (qPCR)-based tests are the gold standard for measuring PVL. Largely because of their high cost, however, implementation of these tests in low- and middle-income countries fails to cover the testing demand. In this study, we aimed at reducing the running cost of the commercially available Abbott RealTime™ HIV-1 assay by minimizing the reagent consumption. To this end, a modified version of the assay was obtained by reducing the assay's reagents volume to about a half, and validated using a panel of 104 plasma samples. Compared to the standard version, the modified Abbott assay allowed for a 50% reduction in running costs. At the same time, it showed a 100% concordance in identifying samples with detectable viral load, strong correlation (Pearson's r=0.983, P<0.0001), and a high agreement between PVL values (mean percent difference between PVL values±standard deviation=0.76±3.18%). In detecting viral failure (PVL>1000copiesmL-1), the modified assay showed a sensitivity of 94.6%, a specificity of 93.8%, and a negative and positive predictive values of 93.8% and 94.6%, respectively. The modified assay therefore reliably quantifies PVL, predicts viral failure, and allows for a ca. 50% reduction in the assay's running costs. It may thus be implemented as an ART monitoring tool in resource-limited settings and for research purposes.

journal_name

J Virol Methods

authors

Boobalan J,Torti A,Dinesha TR,Solomon SS,Balakrishnan P,Saravanan S

doi

10.1016/j.jviromet.2017.05.007

subject

Has Abstract

pub_date

2017-10-01 00:00:00

pages

71-76

eissn

0166-0934

issn

1879-0984

pii

S0166-0934(16)30685-1

journal_volume

248

pub_type

杂志文章
  • Detection of the bovine herpesvirus-1 (BHV-1) genome by PCR.

    abstract::The amplification of the 468 bp fragment of the BHV-1 genome by PCR is described. The 22 bp oligomers from the BHV-1 gI gene were used as primers. For successful amplification the thermal denaturation (100 degrees C/8 min, ice) of the DNA sample was carried out prior to the cycling (95 degrees C for 1 min, 56 degrees ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/0166-0934(93)90132-b

    authors: Vilcek S

    更新日期:1993-02-01 00:00:00

  • Comparison of ELISA and RT-PCR for the detection of Prunus necrotic ring spot virus and prune dwarf virus in almond (Prunus dulcis).

    abstract::A technique based on the reverse transcriptase-polymerase chain reaction (RT-PCR) has been developed to detect the presence of Prunus necrotic ringspot virus (PNRSV) and prune dwarf virus (PDV) simultaneously in almond. This paper presents the results of a 3-year study comparing both enzyme-linked immunosorbent assay ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2003.08.014

    authors: Mekuria G,Ramesh SA,Alberts E,Bertozzi T,Wirthensohn M,Collins G,Sedgley M

    更新日期:2003-12-01 00:00:00

  • Solid phase reverse passive hemadsorption test for hepatitis B surface antigen.

    abstract::A solid phase reverse passive hemadsorption test (SP-RPHAd) for hepatitis B surface antigen detection is described. It was compared with a commercial reverse passive hemagglutination assay (Hepatest, Wellcome, U.K.). SP-RPHAd is four-fold less expensive than Hepatest and undiluted sera can be used instead of eight-fol...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/0166-0934(87)90115-7

    authors: Duverlie G,Driencourt M,Roussel C,Daniel P

    更新日期:1987-11-01 00:00:00

  • Characterisation of visna virus reverse transcriptase: a micro scale reverse transcriptase assay adapted for use with an automated cell harvester.

    abstract::The reverse transcriptase of the sheep lentivirus visna/maedi virus has been characterised. Optima for magnesium ion concentration (5-10 mM), potassium ion concentration (150 mM) and pH (8.25) for this enzyme are very similar to those previously described for the human immunodeficiency viruses. The assay used for this...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/0166-0934(92)90144-3

    authors: Sargan DR,Watt NJ,McConnell I

    更新日期:1992-02-01 00:00:00

  • A comparison of in situ hybridisation, reverse transcriptase-polymerase chain reaction (RT-PCR) and in situ-RT-PCR for the detection of canine distemper virus RNA in Paget's disease.

    abstract::Previous evidence implicating Paramyxoviruses in the aetiopathology of Paget's disease of bone has proved controversial. Whilst several groups have demonstrated Paramyxoviruses using techniques such as in situ hybridisation (ISH), reverse transcriptase-polymerase chain reaction (RT-PCR), and in situ-RT-PCR (IS-RT-PCR)...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/s0166-0934(03)00079-x

    authors: Hoyland JA,Dixon JA,Berry JL,Davies M,Selby PL,Mee AP

    更新日期:2003-05-01 00:00:00

  • Rapid identification of flavivirus using the polymerase chain reaction.

    abstract::A rapid and accurate detection and identification system was developed for flaviviruses that makes use of reverse transcription-polymerase chain reaction (RT-PCR). A primer pair (YF-1 and YF-3), which corresponds to the highly conserved sequence at 3' noncoding region among flaviviruses, was useful for identification ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/0166-0934(93)90020-r

    authors: Tanaka M

    更新日期:1993-03-01 00:00:00

  • The detection of antibodies against foot-and-mouth disease virus in sheep milk.

    abstract::The liquid-phase blocking ELISA (LPBE) and a specific isotype assay (SIA) modified for caprine/ovine IgG1 and IgG2 were used to detect antibodies against foot-and-mouth disease virus isolate O(1) Manisa in sheep milk samples. The majority of samples from animals vaccinated 14-23 weeks previously were indistinguishable...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/s0166-0934(97)00144-4

    authors: Armstrong RM

    更新日期:1997-12-01 00:00:00

  • Serotype-specific detection of adeno-associated virus during laboratory preparation.

    abstract::Adeno-associated virus (AAV) is a small single-stranded DNA member of the family Parvoviradae with at least eight recognized human serotypes, several of which are being studied as candidate vectors for gene therapy. When multiple serotypes are handled in the same laboratory, it is critical to know the serotype of a sa...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2006.05.012

    authors: Mitchell DA,O'Donnell J,Hare JT,Chapman MS

    更新日期:2006-09-01 00:00:00

  • Antigenic characterization of canine parvovirus strains isolated in Italy.

    abstract::28 isolates of canine parvovirus type-2 (CPV-2) were obtained from dogs with hemorrhagic gastroenteritis in Italy. The antigenic structure of CPV-2 isolates was characterized, using four discriminating monoclonal antibodies. In addition, four vaccinal strains were examined. Similar to reports from Australia and the Un...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/s0166-0934(98)00055-x

    authors: Sagazio P,Tempesta M,Buonavoglia D,Cirone F,Buonavoglia C

    更新日期:1998-08-01 00:00:00

  • A method for increasing the surface details of resin embedded viruses using Congo/Crimean haemorrhagic fever virus as a model.

    abstract::The surface details of resin embedded viruses can be greatly enhanced by the use of uranyl acetate block staining. The virions are embedded in agar immediately after fixation and block stained with 3% uranyl acetate in methanol during dehydration. This technique allows those particles outside their host cells to show ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/0166-0934(84)90040-5

    authors: Ellis DS,Smith MD,Lloyd G,Bowen ET,Simpson DI

    更新日期:1984-02-01 00:00:00

  • Novel post-translational modifications of the hemagglutinin and neuraminidase proteins of avian influenza virus expressed by Kluyveromyces lactis.

    abstract::Avian influenza virus (AIV) is an enveloped virus with segmented RNA that belongs to the Orthomyxoviridae. Recently, avian influenza virus isolates have not only posed a significant threat to the poultry industry but also serious public health concerns. The full-length viral hemagglutinin (HA), neuraminidase (NA) or b...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2011.05.006

    authors: Tsai SM,Chiang YC,Chin LT,Liu HJ,Wang CY

    更新日期:2011-08-01 00:00:00

  • Low abundance drug resistance variants in transmitted HIV drug resistance surveillance specimens identified using tagged pooled pyrosequencing.

    abstract::HIV drug resistance (DR) testing using Sanger sequencing (SS) is limited by the inability of the method to identify low abundance drug resistance variants. The application of tagged pooled pyrosequencing (TPP) for HIV DR surveillance is described and the results compared with SS. HIV(+) serum specimens were genotyped ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2012.10.018

    authors: Ji H,Liang B,Li Y,Van Domselaar G,Graham M,Tyler S,Merks H,Sandstrom P,Brooks J

    更新日期:2013-02-01 00:00:00

  • Performance of ViroSeq HIV-1 Genotyping System in routine practice at a Belgian clinical laboratory.

    abstract::Since there are indications of an increasing amount of non-B subtypes in Western Europe it was decided to assess the performance of the ViroSeq HIV-1 Genotyping System on a set of samples from the AIDS Reference Laboratory at the University Hospitals Leuven, a hospital with an increasing number of patients infected wi...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2004.02.005

    authors: Maes B,Schrooten Y,Snoeck J,Derdelinckx I,Van Ranst M,Vandamme AM,Van Laethem K

    更新日期:2004-07-01 00:00:00

  • The creation of stable cell lines expressing Ebola virus glycoproteins and the matrix protein VP40 and generating Ebola virus-like particles utilizing an ecdysone inducible mammalian expression system.

    abstract::Ebola virus is a filovirus that causes hemorrhagic fever in humans and is associated with case fatality rates of up to 90%. The lack of therapeutic interventions in combination with the threat of weaponizing this organism has enhanced research investigations. The expression of key viral proteins and the production of ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2007.12.004

    authors: Melito PL,Qiu X,Fernando LM,deVarennes SL,Beniac DR,Booth TF,Jones SM

    更新日期:2008-03-01 00:00:00

  • A one-step enzyme-linked immunosorbent assay to detect anti-CMV antibodies; development and clinical validation.

    abstract::A simple one-step ELISA to detect anti-CMV antibodies was developed. The test was based on the inhibition principle, and used anti-CMV coated microtitre plates, CMV nuclear antigen and anti-CMV Fab'-HRP conjugate; total assay time was 1.5 h. The use of Fab'-HRP conjugates improved discrimination between positive and n...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/0166-0934(87)90077-2

    authors: Wielaard F,Scherders J,Dagelinckx C,Gausset P

    更新日期:1987-08-01 00:00:00

  • A quantitative real-time reverse transcription PCR (qRT-PCR) assay to detect genome segment 9 of all 26 bluetongue virus serotypes.

    abstract::Bluetongue (BT) is an arboviral disease, which can often be fatal in naïve sheep and white tailed deer, but is usually less severe, or unapparent in other ruminants. Twenty-six bluetongue virus (BTV) serotypes have been recognised so far, two of which (BTV-25 and BTV-26) were recently identified by phylogenetic compar...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2014.11.012

    authors: Maan NS,Maan S,Belaganahalli M,Pullinger G,Montes AJ,Gasparini MR,Guimera M,Nomikou K,Mertens PP

    更新日期:2015-03-01 00:00:00

  • Use of non-radioactively labeled DNA probes for the detection of a baculovirus from Penaeus monodon by in situ hybridization on fixed tissue.

    abstract::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 dist...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/0166-0934(94)90043-4

    authors: Poulos BT,Mari J,Bonami JR,Redman R,Lightner DV

    更新日期:1994-09-01 00:00:00

  • A simple and rapid method for detecting human immunodeficiency virus by PCR.

    abstract::A simple, sensitive and specific method using the polymerase chain reaction (PCR) for amplification of human immunodeficiency virus type 1 (HIV-1) is described. The method involves minimal manipulations. Peripheral blood mononuclear cells (PBMC) were prepared by a rapid Ficoll-Paque gradient method. Lymphocytes were l...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/0166-0934(91)90058-8

    authors: Gibson KM,McLean KA,Clewley JP

    更新日期:1991-05-01 00:00:00

  • A modified immunofluorescence test for Epstein-Barr virus-specific IgM antibody.

    abstract::The fluorescent antibody (FA) test for Epstein-Barr virus (EBV)-specific IgM antibody was improved by the use of sodium butyrate to induce a higher level of EBV antigen expression in P3HR-1 slide preparations and by removal of rheumatoid factor (RF) and IgG antibodies from test sera by means of adsorption with suspens...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/0166-0934(85)90004-7

    authors: Harnett GB,Palmer CA,Bucens MR

    更新日期:1985-10-01 00:00:00

  • Real-time assay for quantitative detection of non-persistently transmitted Plum pox virus RNA targets in single aphids.

    abstract::A TaqMan real-time RT-PCR was developed to detect and quantify RNA-targets from the non-circulative, non-persistently transmitted Plum pox virus (PPV) in individual fresh or aphids captured previously and squashed on paper. Reliable quantitation ranged from 40 up to 4 x 10(8) copies of control transcripts. This techni...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2005.05.011

    authors: Olmos A,Bertolini E,Gil M,Cambra M

    更新日期:2005-09-01 00:00:00

  • Structural and biological properties of Cucumber mosaic virus particles carrying hepatitis C virus-derived epitopes.

    abstract::The Cucumber mosaic virus (CMV) is a three-component isodiametric plant virus with an extremely wide host range, present worldwide. A pseudorecombinant form has been described, deriving from the RNA3 component of the CMV-S strain, carrying the coat protein (CP) gene, and the RNA 1, 2 components of the CMV-D strain. Th...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2008.10.005

    authors: Nuzzaci M,Bochicchio I,De Stradis A,Vitti A,Natilla A,Piazzolla P,Tamburro AM

    更新日期:2009-02-01 00:00:00

  • Application of RT-PCR for the detection of avian reovirus contamination in avian viral vaccines.

    abstract::An efficient procedure for the detection of avian reovirus (ARV)-specific RNA sequences in veterinary immunological medicinal products using reverse transcriptase polymerase chain reaction (RT-PCR) is described. Four ARV vaccine strains (1133, 1733, 2408 and Olson WVU2937), two ATCC strains (VR826 and VR856) as well a...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2004.09.019

    authors: Bruhn S,Bruckner L,Ottiger HP

    更新日期:2005-02-01 00:00:00

  • Restriction analysis of the prototype strain of enteric adenovirus type 41 using exonuclease III.

    abstract::Enteric adenoviruses 40 and 41 (Ad40 and Ad41) are a prominent cause of gastroenteritis in young children. Diagnosis of these enteric types by conventional means is complicated by their fastidious growth characteristics. Enteric adenovirus growth was enhanced by cocultivation. Typing of enteric isolates currently enta...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/0166-0934(92)90166-b

    authors: Scott-Taylor TH,Ahluwalia G,Hammond GW

    更新日期:1992-07-01 00:00:00

  • Diagnosis of measles by fluorescent antibody and culture of nasopharyngeal secretions.

    abstract::An indirect fluorescent antibody test (IFA) was evaluated using commercial mouse anti-measles monoclonal antibody and FITC-labeled goat anti-mouse immunoglobulin. For measles isolation, specimens were inoculated into Rhesus monkey kidney (RMK) cells and, when available, CV-1 cells. 381 specimens were tested by IFA and...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/0166-0934(91)90022-r

    authors: Smaron MF,Saxon E,Wood L,McCarthy C,Morello JA

    更新日期:1991-06-01 00:00:00

  • Three unique Sendai virus antigenic peptides screened from nucleocapsid protein by overlapping peptide array.

    abstract::Sendai virus (SeV) is strictly monitored in laboratory rodents. Currently, complete virions have been used as antigens in SeV serological tests. However, the complexity of SeV virion antigen limits the accuracy of the diagnostic method. In the current study, complete SeV virion antigen was separated on SDS-PAGE and an...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2013.06.011

    authors: Xiang Z,Tong W,Li Y,Qin C,Wei Q

    更新日期:2013-11-01 00:00:00

  • Enzyme-linked fluorescence immunoassays using beta-galactosidase and antibodies covalently bound to polystyrene plates.

    abstract::A solid-phase enzyme-linked immunoassay using a fluorogenic substrate (4-methylumbelliferyl-beta-D-galactopyranoside) was developed. Antibodies were covalently linked to glutaraldehyde-activated 96-well aminopolystyrene plates. Antigens from test samples were adsorbed to the solid phase and detected using antibodies c...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/0166-0934(81)90050-1

    authors: Neurath AR,Strick N

    更新日期:1981-10-01 00:00:00

  • Evaluation of an automated SPF10-LiPA25 assay for detection and typing of human papillomavirus in archival samples.

    abstract::An automated blotter was evaluated for detection of the HPV SPF10-LiPA25 RHA for HPV genotyping with 63 FFPE tissue and 45 WHO proficiency panel samples. The results showed that 10-20% more types were detected and bands were of significantly stronger intensity (p<0.0001) compared to manual processing. Therefore it was...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2014.01.012

    authors: Cornall AM,Quint WH,Garland SM,Tabrizi SN

    更新日期:2014-04-01 00:00:00

  • PG-4 cell plaque assay for xenotropic murine leukemia virus.

    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

    authors: Li Z,Blair M,Thorner L

    更新日期:1999-08-01 00:00:00

  • Comparative detection of rabies RNA by NASBA, real-time PCR and conventional PCR.

    abstract::Five methods for the RNA detection of rabies virus were directly compared in this study. These included conventional nucleic acid sequence-based amplification with electrochemiluminescence (NASBA-ECL) assay, reverse transcription (RT)-heminested (hn) polymerase chain reaction (PCR) and TaqMan real-time RT-PCR using pr...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2011.05.007

    authors: Wacharapluesadee S,Phumesin P,Supavonwong P,Khawplod P,Intarut N,Hemachudha T

    更新日期:2011-08-01 00:00:00

  • The isolation of the two electrophoretic forms of cowpea mosaic virus using fast protein liquid chromatography.

    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

    authors: Smith TJ

    更新日期:1987-07-01 00:00:00