Development of a multiplex real-time reverse transcriptase-polymerase chain reaction for equine infectious anemia virus (EIAV)

R. Frank Cook; S. J. Cook; F. Li; R. C. Montelaro; C. J. Issel

doi:10.1016/S0166-0934(02)00101-5

Development of a multiplex real-time reverse transcriptase-polymerase chain reaction for equine infectious anemia virus (EIAV)

R. Frank Cook, S. J. Cook, F. Li, R. C. Montelaro, C. J. Issel

Research output: Contribution to journal › Article › peer-review

48 Scopus citations

Abstract

A single-tube reverse transcriptase-polymerase chain reaction (RT-PCR) using a fluorogenic real-time PCR detection method is described for the quantitation of equine infectious anemia virus (EIAV) RNA in the plasma of equids. To compensate for variations inherent in sample preparation a multiplex real-time RT-PCR system was developed that permitted the simultaneous calculation of the nucleic acid recovery rate along with the copy number of viral RNA molecules. Detection of EIAV RNA was linear from 10⁹ to 10¹ molecules with intra- and inter-assay variability of less than 1% at 10⁸, 10⁶, 10⁴ and 10² molecules.

Original language	English
Pages (from-to)	171-179
Number of pages	9
Journal	Journal of Virological Methods
Volume	105
Issue number	1
DOIs	https://doi.org/10.1016/S0166-0934(02)00101-5
State	Published - 2002

Bibliographical note

Funding Information:
We thank Diane Furry for preparation of the manuscript. This work was supported by National Institutes of Health grants R01CA49296 and RO1A125850, by funds from the Lucille P. Markey Charitable Trust and the Kentucky Agricultural Experimental Station.

Keywords

Dual-labelled fluorogenic probes
EIAV
Quantitation of viral load
Real-time PCR

ASJC Scopus subject areas

Virology

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10.1016/S0166-0934(02)00101-5

Cite this

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abstract = "A single-tube reverse transcriptase-polymerase chain reaction (RT-PCR) using a fluorogenic real-time PCR detection method is described for the quantitation of equine infectious anemia virus (EIAV) RNA in the plasma of equids. To compensate for variations inherent in sample preparation a multiplex real-time RT-PCR system was developed that permitted the simultaneous calculation of the nucleic acid recovery rate along with the copy number of viral RNA molecules. Detection of EIAV RNA was linear from 109 to 101 molecules with intra- and inter-assay variability of less than 1% at 108, 106, 104 and 102 molecules.",

keywords = "Dual-labelled fluorogenic probes, EIAV, Quantitation of viral load, Real-time PCR",

author = "Cook, {R. Frank} and Cook, {S. J.} and F. Li and Montelaro, {R. C.} and Issel, {C. J.}",

note = "Funding Information: We thank Diane Furry for preparation of the manuscript. This work was supported by National Institutes of Health grants R01CA49296 and RO1A125850, by funds from the Lucille P. Markey Charitable Trust and the Kentucky Agricultural Experimental Station.",

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AU - Cook, S. J.

AU - Li, F.

AU - Montelaro, R. C.

AU - Issel, C. J.

N1 - Funding Information: We thank Diane Furry for preparation of the manuscript. This work was supported by National Institutes of Health grants R01CA49296 and RO1A125850, by funds from the Lucille P. Markey Charitable Trust and the Kentucky Agricultural Experimental Station.

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AB - A single-tube reverse transcriptase-polymerase chain reaction (RT-PCR) using a fluorogenic real-time PCR detection method is described for the quantitation of equine infectious anemia virus (EIAV) RNA in the plasma of equids. To compensate for variations inherent in sample preparation a multiplex real-time RT-PCR system was developed that permitted the simultaneous calculation of the nucleic acid recovery rate along with the copy number of viral RNA molecules. Detection of EIAV RNA was linear from 109 to 101 molecules with intra- and inter-assay variability of less than 1% at 108, 106, 104 and 102 molecules.

KW - Dual-labelled fluorogenic probes

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KW - Quantitation of viral load

KW - Real-time PCR

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Development of a multiplex real-time reverse transcriptase-polymerase chain reaction for equine infectious anemia virus (EIAV)

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

Access to Document

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