Development of a consensus reverse transcription PCR assay for the specific detection of tortoise picornaviruses

Rachel E. Marschang, Katalin Ihász, Renáta Kugler, György Lengyel, Enikő Fehér, Szilvia Marton, K. Bányai, Tara Aqrawi, Szilvia L. Farkas

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Picornaviruses (PVs) of different terrestrial tortoise species, previously designated as Virus “X,” have been frequently detected from various tissues by virus isolation in Terrapene heart cell culture as the preferred laboratory method for diagnosis. Here, we describe the development of 2 diagnostic reverse transcription (RT)-PCR–based assays for the identification and characterization of tortoise PVs belonging to the tentative genus Topivirus. To test the novel diagnostic systems, PVs were isolated from swab and tissue samples collected in Germany, Italy, and Hungary between 2000 and 2013. All 25 tested isolates gave positive results with both novel consensus primer sets. Sequencing of the amplified products confirmed that all studied viruses were members of the new proposed genus Topivirus. Phylogenetic analyses clearly distinguished 2 lineages within the genus. Based on sequence analysis, no association was observed between the geographic distribution and genetic relatedness. Furthermore, no strict host specificity was indicated. The PCR-based diagnosis may provide a time-saving and sensitive method to detect tortoise PVs, and evaluation of PV presence in these animals may help control virus spread.

Original languageEnglish
Pages (from-to)309-314
Number of pages6
JournalJournal of Veterinary Diagnostic Investigation
Volume28
Issue number3
DOIs
Publication statusPublished - 2016

Fingerprint

Picornaviridae
Turtles
tortoises
Reverse Transcription
Consensus
reverse transcriptase polymerase chain reaction
Polymerase Chain Reaction
Viruses
assays
viruses
reverse transcription
Hungary
Clinical Laboratory Techniques
Host Specificity
host specificity
Routine Diagnostic Tests
laboratory techniques
Italy
genetic relationships
Germany

Keywords

  • Diagnostic reverse transcription PCR assay
  • Terrapene heart cell
  • Testudo graeca
  • Topivirus

ASJC Scopus subject areas

  • veterinary(all)

Cite this

Development of a consensus reverse transcription PCR assay for the specific detection of tortoise picornaviruses. / Marschang, Rachel E.; Ihász, Katalin; Kugler, Renáta; Lengyel, György; Fehér, Enikő; Marton, Szilvia; Bányai, K.; Aqrawi, Tara; Farkas, Szilvia L.

In: Journal of Veterinary Diagnostic Investigation, Vol. 28, No. 3, 2016, p. 309-314.

Research output: Contribution to journalArticle

Marschang, Rachel E. ; Ihász, Katalin ; Kugler, Renáta ; Lengyel, György ; Fehér, Enikő ; Marton, Szilvia ; Bányai, K. ; Aqrawi, Tara ; Farkas, Szilvia L. / Development of a consensus reverse transcription PCR assay for the specific detection of tortoise picornaviruses. In: Journal of Veterinary Diagnostic Investigation. 2016 ; Vol. 28, No. 3. pp. 309-314.
@article{6b45d4f00e1749f984fdae49f239e23d,
title = "Development of a consensus reverse transcription PCR assay for the specific detection of tortoise picornaviruses",
abstract = "Picornaviruses (PVs) of different terrestrial tortoise species, previously designated as Virus “X,” have been frequently detected from various tissues by virus isolation in Terrapene heart cell culture as the preferred laboratory method for diagnosis. Here, we describe the development of 2 diagnostic reverse transcription (RT)-PCR–based assays for the identification and characterization of tortoise PVs belonging to the tentative genus Topivirus. To test the novel diagnostic systems, PVs were isolated from swab and tissue samples collected in Germany, Italy, and Hungary between 2000 and 2013. All 25 tested isolates gave positive results with both novel consensus primer sets. Sequencing of the amplified products confirmed that all studied viruses were members of the new proposed genus Topivirus. Phylogenetic analyses clearly distinguished 2 lineages within the genus. Based on sequence analysis, no association was observed between the geographic distribution and genetic relatedness. Furthermore, no strict host specificity was indicated. The PCR-based diagnosis may provide a time-saving and sensitive method to detect tortoise PVs, and evaluation of PV presence in these animals may help control virus spread.",
keywords = "Diagnostic reverse transcription PCR assay, Terrapene heart cell, Testudo graeca, Topivirus",
author = "Marschang, {Rachel E.} and Katalin Ih{\'a}sz and Ren{\'a}ta Kugler and Gy{\"o}rgy Lengyel and Enikő Feh{\'e}r and Szilvia Marton and K. B{\'a}nyai and Tara Aqrawi and Farkas, {Szilvia L.}",
year = "2016",
doi = "10.1177/1040638716628584",
language = "English",
volume = "28",
pages = "309--314",
journal = "Journal of Veterinary Diagnostic Investigation",
issn = "1040-6387",
publisher = "American Association of Veterinary Laboratory Diagnosticians",
number = "3",

}

TY - JOUR

T1 - Development of a consensus reverse transcription PCR assay for the specific detection of tortoise picornaviruses

AU - Marschang, Rachel E.

AU - Ihász, Katalin

AU - Kugler, Renáta

AU - Lengyel, György

AU - Fehér, Enikő

AU - Marton, Szilvia

AU - Bányai, K.

AU - Aqrawi, Tara

AU - Farkas, Szilvia L.

PY - 2016

Y1 - 2016

N2 - Picornaviruses (PVs) of different terrestrial tortoise species, previously designated as Virus “X,” have been frequently detected from various tissues by virus isolation in Terrapene heart cell culture as the preferred laboratory method for diagnosis. Here, we describe the development of 2 diagnostic reverse transcription (RT)-PCR–based assays for the identification and characterization of tortoise PVs belonging to the tentative genus Topivirus. To test the novel diagnostic systems, PVs were isolated from swab and tissue samples collected in Germany, Italy, and Hungary between 2000 and 2013. All 25 tested isolates gave positive results with both novel consensus primer sets. Sequencing of the amplified products confirmed that all studied viruses were members of the new proposed genus Topivirus. Phylogenetic analyses clearly distinguished 2 lineages within the genus. Based on sequence analysis, no association was observed between the geographic distribution and genetic relatedness. Furthermore, no strict host specificity was indicated. The PCR-based diagnosis may provide a time-saving and sensitive method to detect tortoise PVs, and evaluation of PV presence in these animals may help control virus spread.

AB - Picornaviruses (PVs) of different terrestrial tortoise species, previously designated as Virus “X,” have been frequently detected from various tissues by virus isolation in Terrapene heart cell culture as the preferred laboratory method for diagnosis. Here, we describe the development of 2 diagnostic reverse transcription (RT)-PCR–based assays for the identification and characterization of tortoise PVs belonging to the tentative genus Topivirus. To test the novel diagnostic systems, PVs were isolated from swab and tissue samples collected in Germany, Italy, and Hungary between 2000 and 2013. All 25 tested isolates gave positive results with both novel consensus primer sets. Sequencing of the amplified products confirmed that all studied viruses were members of the new proposed genus Topivirus. Phylogenetic analyses clearly distinguished 2 lineages within the genus. Based on sequence analysis, no association was observed between the geographic distribution and genetic relatedness. Furthermore, no strict host specificity was indicated. The PCR-based diagnosis may provide a time-saving and sensitive method to detect tortoise PVs, and evaluation of PV presence in these animals may help control virus spread.

KW - Diagnostic reverse transcription PCR assay

KW - Terrapene heart cell

KW - Testudo graeca

KW - Topivirus

UR - http://www.scopus.com/inward/record.url?scp=84967139357&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84967139357&partnerID=8YFLogxK

U2 - 10.1177/1040638716628584

DO - 10.1177/1040638716628584

M3 - Article

C2 - 27034342

AN - SCOPUS:84967139357

VL - 28

SP - 309

EP - 314

JO - Journal of Veterinary Diagnostic Investigation

JF - Journal of Veterinary Diagnostic Investigation

SN - 1040-6387

IS - 3

ER -