PMID- 33763963
OWN - NLM
STAT- MEDLINE
DCOM- 20220506
LR  - 20220506
IS  - 1865-1682 (Electronic)
IS  - 1865-1674 (Linking)
VI  - 69
IP  - 3
DP  - 2022 May
TI  - Development and validation of a reverse transcription real-time PCR assay for 
      specific detection of PRRSGard vaccine-like virus.
PG  - 1212-1226
LID - 10.1111/tbed.14084 [doi]
AB  - Increasing use of modified live virus (MLV) vaccines presents challenges to 
      interpret positive results of porcine reproductive and respiratory syndrome virus 
      (PRRSV) screening PCR that can detect both wild-type and vaccine strains. 
      Instead, vaccine-specific PCR provides a convenient tool to detect vaccine-like 
      virus from a sample. Here we report the development and validation of a real-time 
      RT-PCR specific for PRRSGard((R)) , a newly available commercial PRRSV-2 MLV 
      vaccine. Analytical specificity, sensitivity and diagnostic performance of 
      PRRSGard PCR were evaluated and compared to a commercial PRRSV screening PCR 
      (reference PCR). PRRSGard and reference PCRs did not cross-react with any of the 
      27 non-PRRSV swine pathogens. PRRSGard PCR did not cross-react with other PRRSV-2 
      vaccine viruses and 31 laboratory and field PRRSV-2 isolates representing various 
      genetic lineages of PRRSV-2. PRRSGard and reference PCRs consistently detected up 
      to 10(-6) and 10(-5) dilutions of PRRSGard vaccine virus, respectively. Based on 
      testing serial dilutions of in vitro transcribed RNA, the 95% limit of detection 
      of PRRSGard PCR was 16 genomic copies/reaction with C(T) cut-off value of 36 and 
      7 genomic copies/reaction with C(T) cut-off value of 37. Diagnostic performance 
      of PRRSGard PCR was evaluated using 846 clinical samples (684 serum and 162 oral 
      fluid samples). Compared to the reference screening PCR, diagnostic sensitivity, 
      specificity and agreement of PRRSGard PCR were 95.34%, 98.85% and 97.52% with 
      cut-off C(T) value of 36 and 98.14%, 96.56% and 97.16% with cut-off C(T) value of 
      37. In addition, PRRSGard PCR was able to detect PRRSGard vaccine virus in a 
      sample even with the co-presence of another PRRSV strain. In summary, in contrast 
      to a reference screening PCR that detects both vaccine and field PRRSV strains, 
      PRRSGard PCR provides a convenient tool to specifically detect PRRSGard 
      vaccine-like virus and to inform PRRSV vaccination protocols.
CI  - (c) 2021 Wiley-VCH GmbH.
FAU - Rawal, Gaurav
AU  - Rawal G
AUID- ORCID: 0000-0003-3753-6119
AD  - Department of Veterinary Diagnostic and Production Animal Medicine, College of 
      Veterinary Medicine, Iowa State University, Ames, IA, USA.
FAU - Yim-Im, Wannarat
AU  - Yim-Im W
AD  - Department of Veterinary Diagnostic and Production Animal Medicine, College of 
      Veterinary Medicine, Iowa State University, Ames, IA, USA.
FAU - Chamba, Fabian
AU  - Chamba F
AD  - Pharmgate Animal Health, Wilmington, NC, USA.
FAU - Smith, Chad
AU  - Smith C
AD  - Pharmgate Animal Health, Wilmington, NC, USA.
FAU - Okones, Jeff
AU  - Okones J
AD  - Pharmgate Animal Health, Wilmington, NC, USA.
FAU - Francisco, Charles
AU  - Francisco C
AD  - Pharmgate Animal Health, Wilmington, NC, USA.
FAU - Zhang, Jianqiang
AU  - Zhang J
AUID- ORCID: 0000-0001-7228-6202
AD  - Department of Veterinary Diagnostic and Production Animal Medicine, College of 
      Veterinary Medicine, Iowa State University, Ames, IA, USA.
LA  - eng
PT  - Journal Article
DEP - 20210427
PL  - Germany
TA  - Transbound Emerg Dis
JT  - Transboundary and emerging diseases
JID - 101319538
RN  - 0 (Vaccines, Attenuated)
RN  - 0 (Viral Vaccines)
SB  - IM
MH  - Animals
MH  - DNA Viruses
MH  - *Porcine Reproductive and Respiratory Syndrome/diagnosis/prevention & control
MH  - *Porcine respiratory and reproductive syndrome virus/genetics
MH  - Real-Time Polymerase Chain Reaction/veterinary
MH  - Reverse Transcription
MH  - Swine
MH  - *Swine Diseases
MH  - Vaccines, Attenuated
MH  - *Viral Vaccines
OTO - NOTNLM
OT  - PRRSGard
OT  - PRRSV
OT  - porcine reproductive and respiratory syndrome virus
OT  - reference screening PCR
OT  - reverse transcription real-time PCR
OT  - vaccine-like
EDAT- 2021/03/26 06:00
MHDA- 2022/05/07 06:00
CRDT- 2021/03/25 06:54
PHST- 2021/03/14 00:00 [revised]
PHST- 2021/01/31 00:00 [received]
PHST- 2021/03/22 00:00 [accepted]
PHST- 2021/03/26 06:00 [pubmed]
PHST- 2022/05/07 06:00 [medline]
PHST- 2021/03/25 06:54 [entrez]
AID - 10.1111/tbed.14084 [doi]
PST - ppublish
SO  - Transbound Emerg Dis. 2022 May;69(3):1212-1226. doi: 10.1111/tbed.14084. Epub 
      2021 Apr 27.