PMID- 35883301
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20220731
IS  - 2076-2615 (Print)
IS  - 2076-2615 (Electronic)
IS  - 2076-2615 (Linking)
VI  - 12
IP  - 14
DP  - 2022 Jul 8
TI  - The Development of a Multiplex Real-Time Quantitative PCR Assay for the 
      Differential Detection of the Wild-Type Strain and the MGF505-2R, EP402R and 
      I177L Gene-Deleted Strain of the African Swine Fever Virus.
LID - 10.3390/ani12141754 [doi]
LID - 1754
AB  - African swine fever virus (ASFV) causes African swine fever (ASF), a devastating 
      hemorrhagic disease of domestic pigs and wild boars. Currently, the MGF505R, 
      EP402R (CD2v) and I177L gene-deleted ASFV strains were confirmed to be the ideal 
      vaccine candidate strains. To develop an assay for differentiating the wild-type 
      and gene-deleted ASFV strains, four pairs of specific primers and TaqMan probes 
      targeting the ASFV B646L (p72), I177L, MGF505-2R and EP402R (CD2v) genes were 
      designed. A multiplex real-time qPCR assay for the differential detection of the 
      wild-type and gene-deleted ASFV strains was developed after optimizing the 
      reaction conditions, including the annealing temperature, primer concentration 
      and probe concentration. The results showed that the multiplex real-time qPCR 
      assay can specifically test the ASFV B646L (p72), I177L, MGF505-2R and EP402R 
      (CD2v) genes with a limit of detection (LOD) of 32.1 copies/muL for the B646L 
      (p72) gene, and 3.21 copies/muL for the I177L, MGF505-2R and EP402R (CD2v) genes. 
      However, the assay cannot test for the classical swine fever virus (CSFV), 
      porcine reproductive and respiratory syndrome virus (PRRSV), porcine epidemic 
      diarrhea virus (PEDV), porcine deltacoronavirus (PDCoV), porcine circovirus type 
      2 (PCV2), PCV3 and pseudorabies virus (PRV). The assay demonstrated good 
      repeatability and reproducibility with coefficients of variation (CV) less than 
      1.56% for both the intra- and inter-assay. The assay was used to test 4239 
      clinical samples, and the results showed that 12.60% (534/4239) samples were 
      positive for ASFV, of which 10 samples lacked the EP402R gene, 6 samples lacked 
      the MGF505-2R gene and 14 samples lacked the EP402R and MGF505-2R genes. The 
      results indicated that the multiplex real-time qPCR developed in this study can 
      provide a rapid, sensitive and specific diagnostic tool for the differential 
      detection of the ASFV B646L, I177L, MGF505-2R and EP402R genes.
FAU - Zhao, Kang
AU  - Zhao K
AUID- ORCID: 0000-0003-3568-8231
AD  - College of Animal Science and Technology, Guangxi University, Nanning 530005, 
      China.
FAU - Shi, Kaichuang
AU  - Shi K
AD  - College of Animal Science and Technology, Guangxi University, Nanning 530005, 
      China.
AD  - Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China.
FAU - Zhou, Qingan
AU  - Zhou Q
AD  - Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China.
FAU - Xiong, Chenyong
AU  - Xiong C
AD  - College of Animal Science and Technology, Guangxi University, Nanning 530005, 
      China.
FAU - Mo, Shenglan
AU  - Mo S
AD  - Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China.
FAU - Zhou, Hongjin
AU  - Zhou H
AD  - College of Animal Science and Technology, Guangxi University, Nanning 530005, 
      China.
FAU - Long, Feng
AU  - Long F
AD  - Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China.
FAU - Wei, Haina
AU  - Wei H
AD  - Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China.
FAU - Hu, Liping
AU  - Hu L
AD  - Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China.
FAU - Mo, Meilan
AU  - Mo M
AUID- ORCID: 0000-0003-3903-0509
AD  - College of Animal Science and Technology, Guangxi University, Nanning 530005, 
      China.
LA  - eng
GR  - AB21238003; AA17204057/the Key Research and Development Program, and the Science 
      and Technology Major Project of Guangxi Science and Technology Bureau, China/
PT  - Journal Article
DEP - 20220708
PL  - Switzerland
TA  - Animals (Basel)
JT  - Animals : an open access journal from MDPI
JID - 101635614
PMC - PMC9311895
OTO - NOTNLM
OT  - African swine fever virus (ASFV)
OT  - detection
OT  - differentiation
OT  - gene-deleted strain
OT  - multiplex real-time qPCR
OT  - wild-type strain
COIS- The authors declare no conflict of interest.
EDAT- 2022/07/28 06:00
MHDA- 2022/07/28 06:01
PMCR- 2022/07/08
CRDT- 2022/07/27 01:05
PHST- 2022/05/31 00:00 [received]
PHST- 2022/07/04 00:00 [revised]
PHST- 2022/07/05 00:00 [accepted]
PHST- 2022/07/27 01:05 [entrez]
PHST- 2022/07/28 06:00 [pubmed]
PHST- 2022/07/28 06:01 [medline]
PHST- 2022/07/08 00:00 [pmc-release]
AID - ani12141754 [pii]
AID - animals-12-01754 [pii]
AID - 10.3390/ani12141754 [doi]
PST - epublish
SO  - Animals (Basel). 2022 Jul 8;12(14):1754. doi: 10.3390/ani12141754.