PMID- 37368767
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20230701
IS  - 2306-7381 (Electronic)
IS  - 2306-7381 (Linking)
VI  - 10
IP  - 6
DP  - 2023 May 31
TI  - Evaluation of a Porcine Endogenous Reference Gene (Internal Sample Control) in a 
      Porcine Reproductive and Respiratory Syndrome Virus RT-qPCR.
LID - 10.3390/vetsci10060381 [doi]
LID - 381
AB  - Endogenous reference genes are used in gene-expression studies to "normalize" the 
      results and, increasingly, as internal sample controls (ISC) in diagnostic 
      quantitative polymerase chain reaction (qPCR). Three studies were conducted to 
      evaluate the performance of a porcine-specific ISC in a commercial porcine 
      reproductive and respiratory syndrome virus (PRRSV) reverse transcription-qPCR. 
      Study 1 evaluated the species specificity of the ISC by testing serum from seven 
      non-porcine domestic species (n = 34). In Study 2, the constancy of ISC detection 
      over time (>/=42 days) was assessed in oral fluid (n = 130), serum (n = 215), and 
      feces (n = 132) collected from individual pigs of known PRRSV status. In Study 3, 
      serum (n = 150), oral fluid (n = 150), and fecal samples (n = 75 feces, 75 fecal 
      swabs) from commercial herds were used to establish ISC reference limits. Study 1 
      showed that the ISC was porcine-specific, i.e., all samples from non-porcine 
      species were ISC negative (n = 34). In Study 2, the ISC was detected in all oral 
      fluid, serum, and fecal samples, but differed in concentration between specimens 
      (p < 0.05; mixed-effects regression model). The results of Study 3 were used to 
      establish ISC reference limits for the 5th, 2.5th and 1.25th percentiles. 
      Overall, the ISC response was consistent to the point that failure in detection 
      is sufficient justification for re-testing and/or re-sampling.
FAU - Munguia-Ramirez, Berenice
AU  - Munguia-Ramirez B
AUID- ORCID: 0000-0002-2557-1817
AD  - Veterinary Diagnostic and Production Animal Medicine Department, College of 
      Veterinary Medicine, Iowa State University, Ames, IA 50011, USA.
FAU - Armenta-Leyva, Betsy
AU  - Armenta-Leyva B
AD  - Veterinary Diagnostic and Production Animal Medicine Department, College of 
      Veterinary Medicine, Iowa State University, Ames, IA 50011, USA.
FAU - Henao-Diaz, Alexandra
AU  - Henao-Diaz A
AD  - Pig Improvement Company (PIC) Mexico, Santiago de Queretaro 76040, Queretaro, 
      Mexico.
FAU - Ye, Fangshu
AU  - Ye F
AUID- ORCID: 0000-0001-6994-1210
AD  - Department of Statistics, College of Liberal Arts and Sciences, Iowa State 
      University, Ames, IA 50011, USA.
FAU - Baum, David H
AU  - Baum DH
AD  - Veterinary Diagnostic and Production Animal Medicine Department, College of 
      Veterinary Medicine, Iowa State University, Ames, IA 50011, USA.
FAU - Gimenez-Lirola, Luis G
AU  - Gimenez-Lirola LG
AUID- ORCID: 0000-0002-4407-7996
AD  - Veterinary Diagnostic and Production Animal Medicine Department, College of 
      Veterinary Medicine, Iowa State University, Ames, IA 50011, USA.
FAU - Zimmerman, Jeffrey J
AU  - Zimmerman JJ
AUID- ORCID: 0000-0002-8107-2422
AD  - Veterinary Diagnostic and Production Animal Medicine Department, College of 
      Veterinary Medicine, Iowa State University, Ames, IA 50011, USA.
LA  - eng
PT  - Journal Article
DEP - 20230531
PL  - Switzerland
TA  - Vet Sci
JT  - Veterinary sciences
JID - 101680127
PMC - PMC10301490
OTO - NOTNLM
OT  - PRRSV
OT  - RT-qPCR
OT  - diagnostics
OT  - endogenous reference genes
OT  - internal sample control
COIS- The authors declare no conflict of interest with respect to the conduct, 
      authorship, and/or publication of this study. Co-author J.J.Z. serves as a 
      consultant to IDEXX Laboratories, Inc. (Westbrook, ME, USA) on areas of 
      diagnostic medicine independent of this research. The terms of the consulting 
      arrangement have been reviewed and approved by Iowa State University in 
      accordance with its conflict-of-interest policies.
EDAT- 2023/06/27 19:13
MHDA- 2023/06/27 19:14
PMCR- 2023/05/31
CRDT- 2023/06/27 12:42
PHST- 2023/05/01 00:00 [received]
PHST- 2023/05/29 00:00 [revised]
PHST- 2023/05/30 00:00 [accepted]
PHST- 2023/06/27 19:14 [medline]
PHST- 2023/06/27 19:13 [pubmed]
PHST- 2023/06/27 12:42 [entrez]
PHST- 2023/05/31 00:00 [pmc-release]
AID - vetsci10060381 [pii]
AID - vetsci-10-00381 [pii]
AID - 10.3390/vetsci10060381 [doi]
PST - epublish
SO  - Vet Sci. 2023 May 31;10(6):381. doi: 10.3390/vetsci10060381.