PMID- 15847915
OWN - NLM
STAT- MEDLINE
DCOM- 20050922
LR  - 20220317
IS  - 0166-0934 (Print)
IS  - 1879-0984 (Electronic)
IS  - 0166-0934 (Linking)
VI  - 126
IP  - 1-2
DP  - 2005 Jun
TI  - The use of endogenous and exogenous reference RNAs for qualitative and 
      quantitative detection of PRRSV in porcine semen.
PG  - 21-30
AB  - Semen is known to be a route of porcine reproductive and respiratory syndrome 
      virus (PRRSV) transmission. A method was developed for qualitative and 
      quantitative detection of the seminal cell-associated PRRSV RNA in relation to 
      endogenous and exogenous reference RNAs. As endogenous control for one-step 
      real-time reverse transcription (RT)-PCR UBE2D2 mRNA was selected. Particularly 
      for the analysis of persistent infections associated with low copy numbers of 
      PRRSV RNA, UBE2D2 mRNA is an ideal control due to its low expression in seminal 
      cells and its detection in all samples analysed (n = 36). However, the amount of 
      UBE2D2 mRNA in porcine semen varied (up to 106-fold), thus its use is limited to 
      qualitative detection of PRRSV RNA. For quantitation, a synthetic, non-metazoan 
      RNA was added to the RNA isolation reaction at an exact copy number. The 
      photosynthesis gene ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit 
      (rbcL) from Arabidopsis thaliana was used as an exogenous spike. Unexpectedly, 
      PRRSV RNA was detected in a herd of specific pathogen-free (SPF) boars which were 
      tested ELISA-negative for anti-PRRSV antibodies. Therefore, RT-PCR for seminal 
      cell-associated PRRSV is a powerful tool for managing the SPF status during 
      quarantine programs and for routine outbreak investigations.
FAU - Revilla-Fernandez, Sandra
AU  - Revilla-Fernandez S
AD  - Institute of Animal Breeding and Genetics, Department for Animal Breeding and 
      Reproduction, University of Veterinary Medicine, Veterinaerplatz 1, A-1210 
      Vienna, Austria.
FAU - Wallner, Barbara
AU  - Wallner B
FAU - Truschner, Klaus
AU  - Truschner K
FAU - Benczak, Alexandra
AU  - Benczak A
FAU - Brem, Gottfried
AU  - Brem G
FAU - Schmoll, Friedrich
AU  - Schmoll F
FAU - Mueller, Mathias
AU  - Mueller M
FAU - Steinborn, Ralf
AU  - Steinborn R
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (RNA, Messenger)
RN  - 0 (RNA, Viral)
RN  - EC 2.3.2.23 (Ubiquitin-Conjugating Enzymes)
RN  - EC 4.1.1.39 (RbcL protein, plastid)
RN  - EC 4.1.1.39 (Ribulose-Bisphosphate Carboxylase)
SB  - IM
MH  - Animals
MH  - Molecular Sequence Data
MH  - Porcine Reproductive and Respiratory Syndrome/*diagnosis/virology
MH  - Porcine respiratory and reproductive syndrome virus/genetics/*isolation & 
      purification
MH  - RNA, Messenger/analysis
MH  - RNA, Viral/*analysis/genetics
MH  - Reference Standards
MH  - Reverse Transcriptase Polymerase Chain Reaction/*standards
MH  - Ribulose-Bisphosphate Carboxylase/genetics
MH  - Semen/*virology
MH  - Sensitivity and Specificity
MH  - Sequence Analysis, DNA
MH  - Swine
MH  - Ubiquitin-Conjugating Enzymes/genetics
PMC - PMC7112884
EDAT- 2005/04/26 09:00
MHDA- 2005/09/24 09:00
PMCR- 2005/02/23
CRDT- 2005/04/26 09:00
PHST- 2004/08/12 00:00 [received]
PHST- 2005/01/17 00:00 [revised]
PHST- 2005/01/25 00:00 [accepted]
PHST- 2005/04/26 09:00 [pubmed]
PHST- 2005/09/24 09:00 [medline]
PHST- 2005/04/26 09:00 [entrez]
PHST- 2005/02/23 00:00 [pmc-release]
AID - S0166-0934(05)00030-3 [pii]
AID - 10.1016/j.jviromet.2005.01.018 [doi]
PST - ppublish
SO  - J Virol Methods. 2005 Jun;126(1-2):21-30. doi: 10.1016/j.jviromet.2005.01.018.