A versatile qpcr for diagnosis of leporid gammaherpesvirus 5 using evagreen® or taqman® technologies

Fábio A. Abade Dos Santos; Carina L. Carvalho; Maria C. Peleteiro; Francisco Parra; Margarida D. Duarte

doi:10.3390/v13040715

A versatile qpcr for diagnosis of leporid gammaherpesvirus 5 using evagreen® or taqman® technologies

Fábio A. Abade Dos Santos, Carina L. Carvalho, Maria C. Peleteiro, Francisco Parra, Margarida D. Duarte

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

5 Citations (Scopus)

Abstract

In late 2019, the first herpesvirus in the genus Lepus, named leporid gammaherpesvirus 5 (LeHV-5) was described. At the time, herpetic typical lesions were observed in hares infected by the myxoma virus, which is known to induce immunosuppression. Though the real impact of LeHV-5 is still poorly understood, since it affects reproduction, it poses an additional threat to the already fragile populations of Iberian hare, demanding prevalence investigations. In this article, we describe the first quantitative molecular method for LeHV-5 detection, using either Taqman or the EvaGreen systems. This method has excellent sensitivity and specificity, it is able to detect 2.1 copies of LeHV-5 DNA and was validated with an internal control targeting the 18S rRNA gene, allowing monitoring extraction and PCR amplification efficiencies.

Original language	English
Article number	715
Journal	Viruses
Volume	13
Issue number	4
DOIs	https://doi.org/10.3390/v13040715
Publication status	Published - Apr 2021
Externally published	Yes

Keywords

Gammaherpesvirus
Herpesvirus
Iberian hare
Leporid gammaherpesvirus 5
Lepus granatensis
QPCR
Reproduction

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10.3390/v13040715

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title = "A versatile qpcr for diagnosis of leporid gammaherpesvirus 5 using evagreen{\textregistered} or taqman{\textregistered} technologies",

abstract = "In late 2019, the first herpesvirus in the genus Lepus, named leporid gammaherpesvirus 5 (LeHV-5) was described. At the time, herpetic typical lesions were observed in hares infected by the myxoma virus, which is known to induce immunosuppression. Though the real impact of LeHV-5 is still poorly understood, since it affects reproduction, it poses an additional threat to the already fragile populations of Iberian hare, demanding prevalence investigations. In this article, we describe the first quantitative molecular method for LeHV-5 detection, using either Taqman or the EvaGreen systems. This method has excellent sensitivity and specificity, it is able to detect 2.1 copies of LeHV-5 DNA and was validated with an internal control targeting the 18S rRNA gene, allowing monitoring extraction and PCR amplification efficiencies.",

keywords = "Gammaherpesvirus, Herpesvirus, Iberian hare, Leporid gammaherpesvirus 5, Lepus granatensis, QPCR, Reproduction",

author = "{Abade Dos Santos}, {F{\'a}bio A.} and Carvalho, {Carina L.} and Peleteiro, {Maria C.} and Francisco Parra and Duarte, {Margarida D.}",

note = "Publisher Copyright: {\textcopyright} 2021 by the authors. Licensee MDPI, Basel, Switzerland.",

year = "2021",

month = apr,

doi = "10.3390/v13040715",

language = "English",

volume = "13",

journal = "Viruses",

issn = "1999-4915",

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T1 - A versatile qpcr for diagnosis of leporid gammaherpesvirus 5 using evagreen® or taqman® technologies

AU - Abade Dos Santos, Fábio A.

AU - Carvalho, Carina L.

AU - Peleteiro, Maria C.

AU - Parra, Francisco

AU - Duarte, Margarida D.

PY - 2021/4

Y1 - 2021/4

N2 - In late 2019, the first herpesvirus in the genus Lepus, named leporid gammaherpesvirus 5 (LeHV-5) was described. At the time, herpetic typical lesions were observed in hares infected by the myxoma virus, which is known to induce immunosuppression. Though the real impact of LeHV-5 is still poorly understood, since it affects reproduction, it poses an additional threat to the already fragile populations of Iberian hare, demanding prevalence investigations. In this article, we describe the first quantitative molecular method for LeHV-5 detection, using either Taqman or the EvaGreen systems. This method has excellent sensitivity and specificity, it is able to detect 2.1 copies of LeHV-5 DNA and was validated with an internal control targeting the 18S rRNA gene, allowing monitoring extraction and PCR amplification efficiencies.

AB - In late 2019, the first herpesvirus in the genus Lepus, named leporid gammaherpesvirus 5 (LeHV-5) was described. At the time, herpetic typical lesions were observed in hares infected by the myxoma virus, which is known to induce immunosuppression. Though the real impact of LeHV-5 is still poorly understood, since it affects reproduction, it poses an additional threat to the already fragile populations of Iberian hare, demanding prevalence investigations. In this article, we describe the first quantitative molecular method for LeHV-5 detection, using either Taqman or the EvaGreen systems. This method has excellent sensitivity and specificity, it is able to detect 2.1 copies of LeHV-5 DNA and was validated with an internal control targeting the 18S rRNA gene, allowing monitoring extraction and PCR amplification efficiencies.

KW - Gammaherpesvirus

KW - Herpesvirus

KW - Iberian hare

KW - Leporid gammaherpesvirus 5

KW - Lepus granatensis

KW - QPCR

KW - Reproduction

UR - http://www.scopus.com/inward/record.url?scp=85105209643&partnerID=8YFLogxK

U2 - 10.3390/v13040715

DO - 10.3390/v13040715

M3 - Article

C2 - 33924254

AN - SCOPUS:85105209643

SN - 1999-4915

VL - 13

JO - Viruses

JF - Viruses

IS - 4

M1 - 715

ER -

A versatile qpcr for diagnosis of leporid gammaherpesvirus 5 using evagreen® or taqman® technologies

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Keywords

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