Development and application of a SYBR green RT-PCR for first line screening and quantification of porcine sapovirus infection

A. Mauroy, W.H.M. van der Poel, R.W. van der Honing-Hakze, C. Thys, E. Thiry

Research output: Contribution to journalArticleAcademicpeer-review

5 Citations (Scopus)

Abstract

Background: Sapoviruses are single stranded positive sense RNA viruses belonging to the family Caliciviridae. The virus is detected in different species including the human and the porcine species as an enteric pathogen causing asymptomatic to symptomatic enteritis. In this study, we report the development of a rapid real time qRT-PCR based on SYBR Green chemistry for the diagnosis of porcine sapovirus infection in swine. Results: The method allows the detection of porcine sapoviruses and the quantification of the genomic copies present in stool samples. During its development, the diagnostic tool showed good correlation compared with the gold standard conventional RT-PCR and was ten-fold more sensitive. When the method was applied to field samples, porcine noroviruses from genogroup 2 genotype 11b were also detected. The method was also applied to swine samples from the Netherlands that were positive for PoSaV infection. Phylogenetic results obtained from the samples showed that PoSaV sequences were genetically related to the currently described genogroup III, to the proposed genogroup VII and also to the MI-QW19 sequence (close to the human SaV sequences). Conclusions: A rapid, sensitive, and reliable diagnosis method was developed for porcine sapovirus diagnosis. It correlated with the gold standard conventional RT-PCR. Specificity was good apart for genogroup 2 genotype 11b porcine noroviruses. As a first line screening diagnosis method, it allows a quicker and easier decision on doubtful samples.
Original languageEnglish
Article number193
Number of pages11
JournalBMC Veterinary Research
Volume8
DOIs
Publication statusPublished - 2012

Fingerprint

Sapovirus
Swine
reverse transcriptase polymerase chain reaction
screening
Polymerase Chain Reaction
swine
Genotype
Infection
infection
Norovirus
gold
positive sense, single-stranded RNA viruses
Caliciviridae
sampling
green chemistry
methodology
Enteritis
genotype
RNA Viruses
enteritis

Keywords

  • virus-like particles
  • genetic diversity
  • enteric caliciviruses
  • epidemiology
  • noroviruses
  • recombination
  • pigs
  • classification
  • europe
  • assays

Cite this

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title = "Development and application of a SYBR green RT-PCR for first line screening and quantification of porcine sapovirus infection",
abstract = "Background: Sapoviruses are single stranded positive sense RNA viruses belonging to the family Caliciviridae. The virus is detected in different species including the human and the porcine species as an enteric pathogen causing asymptomatic to symptomatic enteritis. In this study, we report the development of a rapid real time qRT-PCR based on SYBR Green chemistry for the diagnosis of porcine sapovirus infection in swine. Results: The method allows the detection of porcine sapoviruses and the quantification of the genomic copies present in stool samples. During its development, the diagnostic tool showed good correlation compared with the gold standard conventional RT-PCR and was ten-fold more sensitive. When the method was applied to field samples, porcine noroviruses from genogroup 2 genotype 11b were also detected. The method was also applied to swine samples from the Netherlands that were positive for PoSaV infection. Phylogenetic results obtained from the samples showed that PoSaV sequences were genetically related to the currently described genogroup III, to the proposed genogroup VII and also to the MI-QW19 sequence (close to the human SaV sequences). Conclusions: A rapid, sensitive, and reliable diagnosis method was developed for porcine sapovirus diagnosis. It correlated with the gold standard conventional RT-PCR. Specificity was good apart for genogroup 2 genotype 11b porcine noroviruses. As a first line screening diagnosis method, it allows a quicker and easier decision on doubtful samples.",
keywords = "virus-like particles, genetic diversity, enteric caliciviruses, epidemiology, noroviruses, recombination, pigs, classification, europe, assays",
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Development and application of a SYBR green RT-PCR for first line screening and quantification of porcine sapovirus infection. / Mauroy, A.; van der Poel, W.H.M.; van der Honing-Hakze, R.W.; Thys, C.; Thiry, E.

In: BMC Veterinary Research, Vol. 8, 193, 2012.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Development and application of a SYBR green RT-PCR for first line screening and quantification of porcine sapovirus infection

AU - Mauroy, A.

AU - van der Poel, W.H.M.

AU - van der Honing-Hakze, R.W.

AU - Thys, C.

AU - Thiry, E.

N1 - WOS:000312670600001

PY - 2012

Y1 - 2012

N2 - Background: Sapoviruses are single stranded positive sense RNA viruses belonging to the family Caliciviridae. The virus is detected in different species including the human and the porcine species as an enteric pathogen causing asymptomatic to symptomatic enteritis. In this study, we report the development of a rapid real time qRT-PCR based on SYBR Green chemistry for the diagnosis of porcine sapovirus infection in swine. Results: The method allows the detection of porcine sapoviruses and the quantification of the genomic copies present in stool samples. During its development, the diagnostic tool showed good correlation compared with the gold standard conventional RT-PCR and was ten-fold more sensitive. When the method was applied to field samples, porcine noroviruses from genogroup 2 genotype 11b were also detected. The method was also applied to swine samples from the Netherlands that were positive for PoSaV infection. Phylogenetic results obtained from the samples showed that PoSaV sequences were genetically related to the currently described genogroup III, to the proposed genogroup VII and also to the MI-QW19 sequence (close to the human SaV sequences). Conclusions: A rapid, sensitive, and reliable diagnosis method was developed for porcine sapovirus diagnosis. It correlated with the gold standard conventional RT-PCR. Specificity was good apart for genogroup 2 genotype 11b porcine noroviruses. As a first line screening diagnosis method, it allows a quicker and easier decision on doubtful samples.

AB - Background: Sapoviruses are single stranded positive sense RNA viruses belonging to the family Caliciviridae. The virus is detected in different species including the human and the porcine species as an enteric pathogen causing asymptomatic to symptomatic enteritis. In this study, we report the development of a rapid real time qRT-PCR based on SYBR Green chemistry for the diagnosis of porcine sapovirus infection in swine. Results: The method allows the detection of porcine sapoviruses and the quantification of the genomic copies present in stool samples. During its development, the diagnostic tool showed good correlation compared with the gold standard conventional RT-PCR and was ten-fold more sensitive. When the method was applied to field samples, porcine noroviruses from genogroup 2 genotype 11b were also detected. The method was also applied to swine samples from the Netherlands that were positive for PoSaV infection. Phylogenetic results obtained from the samples showed that PoSaV sequences were genetically related to the currently described genogroup III, to the proposed genogroup VII and also to the MI-QW19 sequence (close to the human SaV sequences). Conclusions: A rapid, sensitive, and reliable diagnosis method was developed for porcine sapovirus diagnosis. It correlated with the gold standard conventional RT-PCR. Specificity was good apart for genogroup 2 genotype 11b porcine noroviruses. As a first line screening diagnosis method, it allows a quicker and easier decision on doubtful samples.

KW - virus-like particles

KW - genetic diversity

KW - enteric caliciviruses

KW - epidemiology

KW - noroviruses

KW - recombination

KW - pigs

KW - classification

KW - europe

KW - assays

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DO - 10.1186/1746-6148-8-193

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JO - BMC Veterinary Research

JF - BMC Veterinary Research

SN - 1746-6148

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ER -