PMID- 34854269
OWN - NLM
STAT- MEDLINE
DCOM- 20220203
LR  - 20220203
IS  - 1976-555X (Electronic)
IS  - 1229-845X (Print)
IS  - 1229-845X (Linking)
VI  - 22
IP  - 6
DP  - 2021 Nov
TI  - Development of a multiplex qRT-PCR assay for detection of African swine fever 
      virus, classical swine fever virus and porcine reproductive and respiratory 
      syndrome virus.
PG  - e87
LID - 10.4142/jvs.2021.22.e87 [doi]
LID - e87
AB  - BACKGROUND: African swine fever virus (ASFV), classical swine fever virus (CSFV), 
      and porcine reproductive and respiratory syndrome virus (PRRSV) are still 
      prevalent in many regions of China. Co-infections make it difficult to 
      distinguish their clinical symptoms and pathological changes. Therefore, a rapid 
      and specific method is needed for the differential detection of these pathogens. 
      OBJECTIVES: The aim of this study was to develop a multiplex real-time 
      quantitative reverse transcription polymerase chain reaction (multiplex qRT-PCR) 
      for the simultaneous differential detection of ASFV, CSFV, and PRRSV. METHODS: 
      Three pairs of primers and TaqMan probes targeting the ASFV p72 gene, CSFV 5' 
      untranslated region, and PRRSV ORF7 gene were designed. After optimizing the 
      reaction conditions, including the annealing temperature, primer concentration, 
      and probe concentration, multiplex qRT-PCR for simultaneous and differential 
      detection of ASFV, CSFV, and PRRSV was developed. Subsequently, 1,143 clinical 
      samples were detected to verify the practicality of the assay. RESULTS: The 
      multiplex qRT-PCR assay could specifically and simultaneously detect the ASFV, 
      CSFV, and PRRSV with a detection limit of 1.78 x 10(0) copies for the ASFV, CSFV, 
      and PRRSV, but could not amplify the other major porcine viruses, such as 
      pseudorabies virus, porcine circovirus type 1 (PCV1), PCV2, PCV3, foot-and-mouth 
      disease virus, porcine parvovirus, atypical porcine pestivirus, and Senecavirus 
      A. The assay had good repeatability with coefficients of variation of intra- and 
      inter-assay of less than 1.2%. Finally, the assay was used to detect 1,143 
      clinical samples to evaluate its practicality in the field. The positive rates of 
      ASFV, CSFV, and PRRSV were 25.63%, 9.36%, and 17.50%, respectively. The 
      co-infection rates of ASFV+CSFV, ASFV+PRRSV, CSFV+PRRSV, and ASFV+CSFV+PRRSV were 
      2.45%, 2.36%, 1.57%, and 0.17%, respectively. CONCLUSIONS: The multiplex qRT-PCR 
      developed in this study could provide a rapid, sensitive, specific diagnostic 
      tool for the simultaneous and differential detection of ASFV, CSFV, and PRRSV.
CI  - (c) 2021 The Korean Society of Veterinary Science.
FAU - Chen, Yating
AU  - Chen Y
AUID- ORCID: 0000-0001-5510-960X
AD  - College of Animal Science and Technology, Guangxi University, Nanning 530005, 
      China.
FAU - Shi, Kaichuang
AU  - Shi K
AUID- ORCID: 0000-0002-1023-9635
AD  - College of Animal Science and Technology, Guangxi University, Nanning 530005, 
      China.
AD  - Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China. 
      shikaichuang@126.com.
FAU - Liu, Huixin
AU  - Liu H
AUID- ORCID: 0000-0001-5492-1209
AD  - College of Animal Science and Technology, Guangxi University, Nanning 530005, 
      China.
FAU - Yin, Yanwen
AU  - Yin Y
AUID- ORCID: 0000-0002-3890-2080
AD  - Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China.
FAU - Zhao, Jing
AU  - Zhao J
AUID- ORCID: 0000-0001-7771-7542
AD  - College of Animal Science and Technology, Guangxi University, Nanning 530005, 
      China.
FAU - Long, Feng
AU  - Long F
AUID- ORCID: 0000-0002-9625-0527
AD  - Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China.
FAU - Lu, Wenjun
AU  - Lu W
AUID- ORCID: 0000-0003-2989-045X
AD  - Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China.
FAU - Si, Hongbin
AU  - Si H
AUID- ORCID: 0000-0002-0416-6150
AD  - College of Animal Science and Technology, Guangxi University, Nanning 530005, 
      China. shb2009@gxu.edu.cn.
LA  - eng
GR  - AA17204057/Scientific Research and Technology Development Program of 
      Guangxi/China
GR  - Z201954/Guangxi Agricultural and Rural Bureau/China
GR  - Z202031/Guangxi Agricultural and Rural Bureau/China
PT  - Journal Article
PL  - Korea (South)
TA  - J Vet Sci
JT  - Journal of veterinary science
JID - 100964185
SB  - IM
MH  - *African Swine Fever Virus/isolation & purification
MH  - Animals
MH  - China/epidemiology
MH  - *Classical Swine Fever Virus/isolation & purification
MH  - *Porcine respiratory and reproductive syndrome virus/isolation & purification
MH  - Real-Time Polymerase Chain Reaction/*veterinary
MH  - Swine
PMC - PMC8636662
OTO - NOTNLM
OT  - African swine fever virus
OT  - classical swine fever virus
OT  - multiplex real-time quantitative RT-PCR (multiplex qRT-PCR)
OT  - porcine reproductive and respiratory syndrome virus
COIS- The authors declare no conflicts of interest.
EDAT- 2021/12/03 06:00
MHDA- 2022/02/04 06:00
PMCR- 2021/10/01
CRDT- 2021/12/02 07:02
PHST- 2021/05/05 00:00 [received]
PHST- 2021/08/04 00:00 [revised]
PHST- 2021/08/12 00:00 [accepted]
PHST- 2021/12/02 07:02 [entrez]
PHST- 2021/12/03 06:00 [pubmed]
PHST- 2022/02/04 06:00 [medline]
PHST- 2021/10/01 00:00 [pmc-release]
AID - 22.e87 [pii]
AID - 10.4142/jvs.2021.22.e87 [doi]
PST - ppublish
SO  - J Vet Sci. 2021 Nov;22(6):e87. doi: 10.4142/jvs.2021.22.e87.