Quantitative analysis of maedi-visna virus DNA load in peripheral blood monocytes and alveolar macrophages

Zhang, Zhidong; Watt, N.J.; Harkiss, G. D.; Woodall, Chris

doi:10.1016/s0166-0934(99)00169-x

Cited by 49 publications

(48 citation statements)

References 10 publications

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“…The low viral load in blood is a major challenge for virus detection. In addition, as shown in goats experimentally infected with CAEV and in sheep infected with MVV, the viral load may fluctuate over time and may differ between individual animals [70,156].…”

Section: Virus Detectionmentioning

confidence: 99%

Routes of transmission and consequences of small ruminant lentiviruses (SRLVs) infection and eradication schemes

et al. 2004

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-Small ruminant lentiviruses (SRLV = maedi-visna in sheep and caprine arthritis encephalitis in goats) are distributed throughout most countries of the world, particularly Europe. Laboratories from 16 European countries established collaborations within the framework of a COST (CO-operation in the field of Scientific and Technical Research) action sponsored by the European Union in order to (i) better organize their research programmes on SRLVs and (ii) to coordinate efforts to combat these two diseases. After five years, a consensus conference -the first one in the veterinary medicine field -concluded the work of this network of laboratories by reviewing the present position and discussing three important questions in the field of SRLVs: routes of transmission, consequences of infection and potential role of eradication programmes at either a European or local level, according to the situation in each country or region. This paper brings together existing information regarding these questions and identifies areas for future research.Maedi-visna / caprine arthritis-encephalitis virus / lentivirus / small ruminants / European consensus conference

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Section: Virus Detectionmentioning

confidence: 99%

Routes of transmission and consequences of small ruminant lentiviruses (SRLVs) infection and eradication schemes

et al. 2004

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“…Several in vivo and in vitro models of infection with EV1 have been established over the years, where the viral sequences were detected mostly by nested PCR, allowing only approximate quantitative evaluations ( Bird et al, 1993and Ryan et al, 2000. A QC PCR was developed to determine accurately and compare the EV1 viral load in peripheral blood monocytes and in alveolar macrophages of infected sheep ( Zhang et al, 2000). However this technique is quite laborious and not suited for the analysis of large numbers of samples.…”

Section: Discussionmentioning

confidence: 99%

“…of 0.4 TCID50/cell. Two days later genomic DNA was extracted (DNeasy Tissue Kit, Qiagen, Hilden, Germany) and the presence of EV1 provirus was confirmed by nested pol PCR ( Zhang et al, 2000).…”

Section: Cloning and Sequencing Of Ev1 Gag And Pol Gene Fragmentsmentioning

confidence: 99%

“…This strain has been and is currently used for in vivo and in vitro studies focusing on diverse topics such as the identification of viral promoter elements, or cytokines that control the expression and replication of the virus , Sutton et al, 1997and Zhang et al, 2002, the dissection of the initial steps of the infection ( Bird et al, 1993, Blacklaws et al, 1995and Ryan et al, 2000, the characterization of the immune response ( Bird et al, 1993 andSingh et al, 2006), the routes of viral entry and dissemination ( Blacklaws et al, 1995, McNeilly et al, 2007 and the development of immunization strategies ( Gonzalez et al, 2005, Niesalla et al, 2009and Reina et al, 2008. In some of these studies the presence of the EV1 genome has been investigated by conventional PCR ( Bird et al, 1993and Ryan et al, 2000 or by quantitative competitive PCR (QC PCR) ( Zhang et al, 2000). The aim of this work was the development of a highly sensitive dual labelled probe real--time PCR for the absolute quantitation of EV1 viral load in vivo and in vitro.…”

Section: Introductionmentioning

confidence: 99%

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Development and comparison of strain specific gag and pol real-time PCR assays for the detection of Visna/maedi virus

Carrozza

Mazzei

Bandecchi

et al. 2010

Journal of Virological Methods

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The aim of this study was the development of gag and pol dual labelled probe real-time PCR and RT PCR assays to quantify the proviral load and the transcripts of the British Visna/maedi virus EV1 strain. Primers and probes were chosen based on the consensus sequences of gag and pol clones representative of EV1 genetic variants. Both PCRs had a detection limit of 3 copies of target gene, with a linearity over 6 orders of magnitude. The performances of the two PCRs in vivo were evaluated and compared on a panel of DNAs extracted from blood of sheep infected experimentally with EV1. The pol assay detected in most cases lower numbers of viral molecules than gag assay, yielding some false negative results. The gag real-time RT PCR had a detection limit of 100 RNA molecules with a linearity over 5 orders of magnitude. This did not result in a lower performance of the RT PCR compared to the PCR in cells permissive for virus replication, which contain higher numbers of viral transcripts than proviral genomes. The real-time assays developed in this study, particularly the gag assay, provide a sensitive tool which can be used to quantify the viral load in experimental infections

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“…SRLV are widely spread and strategies based on early diagnosis, management and culling of seropositive animals have been applied in order to eradicate infection. In natural SRLV infections of small ruminants, the response of the immune system to virus in tissues may lead to pathology [2], [3], [4], [5] and [6], which may increase in heavily infected animals [2], [7] and [8] till the animal's death. Although vaccination may prevent infection, minimize clinical symptoms or delay the onset of disease resulting in an improvement of animal welfare and avoidance of production losses (reviewed in Ref.…”

Section: Introductionmentioning

confidence: 99%

Use of B7 costimulatory molecules as adjuvants in a prime-boost vaccination against Visna/Maedi ovine lentivirus

Andrés

Reina

Ciriza

et al. 2009

Vaccine

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RNA transcripts of the B7 family molecule (CD80) are diminished in blood leukocytes from animals clinically affected with Visna/Maedi virus (VMV) infection. This work investigates whether the use of B7 genes enhances immune responses and protection in immunization--challenge approaches. Sheep were primed by particle--mediated epidermal bombardment with VMV gag and env gene recombinant plasmids together with plasmids encoding both CD80 and CD86 or CD80 alone, boosted with gag and env gene recombinant modified vaccinia Ankara virus and challenged intratracheally with VMV. Immunization in the presence of one or both of the B7 genes resulted in CD4+ T cell activation and antibody production (before and after challenge, respectively), but only immunization with CD80 and CD86 genes together, and not CD80 alone, resulted in a reduced number of infected animals and increased early transient cytotoxic T lymphocytes (CTL) responses. Post--mortem analysis showed an immune activation of lymphoid tissue in challenge--target organs in those animals that had received B7 genes compared to unvaccinated animals. Thus, the inclusion of B7 genes helped to enhance early cellular responses and protection (diminished proportion of infected animals) against VMV infection.

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Quantitative analysis of maedi-visna virus DNA load in peripheral blood monocytes and alveolar macrophages

Cited by 49 publications

References 10 publications

Routes of transmission and consequences of small ruminant lentiviruses (SRLVs) infection and eradication schemes

Routes of transmission and consequences of small ruminant lentiviruses (SRLVs) infection and eradication schemes

Development and comparison of strain specific gag and pol real-time PCR assays for the detection of Visna/maedi virus

Use of B7 costimulatory molecules as adjuvants in a prime-boost vaccination against Visna/Maedi ovine lentivirus

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