PMID- 35042532
OWN - NLM
STAT- MEDLINE
DCOM- 20220303
LR  - 20220303
IS  - 1746-6148 (Electronic)
IS  - 1746-6148 (Linking)
VI  - 18
IP  - 1
DP  - 2022 Jan 18
TI  - Development of a one-step multiplex qRT-PCR assay for the detection of African 
      swine fever virus, classical swine fever virus and atypical porcine pestivirus.
PG  - 43
LID - 10.1186/s12917-022-03144-4 [doi]
LID - 43
AB  - BACKGROUND: African swine fever virus (ASFV), classical swine fever virus (CSFV) 
      and atypical porcine pestivirus (APPV) have caused great economic losses to the 
      swine industry in China. Since coinfections of ASFV, CSFV and APPV occur in 
      certain pig herds, it is necessary to accurately and differentially detect these 
      pathogens in field-collected samples. In this study, a one-step multiplex 
      real-time quantitative reverse transcription-polymerase chain reaction (multiplex 
      qRT-PCR) was developed for the simultaneous and differential detection of ASFV, 
      CSFV and APPV. RESULTS: The one-step multiplex qRT-PCR presented here was able to 
      simultaneously detect ASFV, CSFV and APPV but could not amplify other viruses, 
      including porcine circovirus type 2 (PCV2), pseudorabies virus (PRV), porcine 
      reproductive and respiratory syndrome virus (PRRSV), foot-and-mouth disease virus 
      (FMDV), porcine parvovirus (PPV), porcine epidemic diarrhoea virus (PEDV), 
      transmissible gastroenteritis virus (TGEV), porcine rotavirus (PRoV), porcine 
      deltacoronavirus (PDCoV), border disease virus (BDV), bovine viral diarrhoea 
      virus type 1 (BVDV-1), BVDV-2, etc. The limit of detection (LOD) of the assay was 
      2.52 x 10(1) copies/muL for ASFV, CSFV and APPV. A repeatability test using 
      standard recombinant plasmids showed that the intra- and interassay coefficients 
      of variation (CVs) were less than 2%. An assay of 509 clinical samples collected 
      in Guangxi Province, southern China, from October 2018 to December 2020 showed 
      that the positive rates of ASFV, CSFV and APPV were 45.58, 12.57 and 3.54%, 
      respectively, while the coinfection rates of ASFV and CSFV, ASFV and APPV, CSFV 
      and APPV were 4.91, 1.38, 0.98%, respectively. Phylogenetic analysis based on the 
      nucleotide sequences of the partial ASFV p72 gene showed that all ASFV strains 
      from Guangxi Province belonged to genotypes I and II. CONCLUSION: A one-step 
      multiplex qRT-PCR with high specificity, sensitivity and repeatability was 
      successfully developed for the simultaneous and differential detection of ASFV, 
      CSFV and APPV.
CI  - (c) 2022. The Author(s).
FAU - Liu, Huixin
AU  - Liu H
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. shikaichuang@126.com.
AD  - Guangxi Center for Animal Disease Control and Prevention, Nanning, 530001, China. 
      shikaichuang@126.com.
FAU - Zhao, Jing
AU  - Zhao J
AD  - College of Animal Science and Technology, Guangxi University, Nanning, 530005, 
      China.
FAU - Yin, Yanwen
AU  - Yin Y
AD  - Guangxi Center for Animal Disease Control and Prevention, Nanning, 530001, China.
FAU - Chen, Yating
AU  - Chen Y
AD  - College of Animal Science and Technology, Guangxi University, Nanning, 530005, 
      China.
FAU - Si, Hongbin
AU  - Si H
AD  - College of Animal Science and Technology, Guangxi University, Nanning, 530005, 
      China. shb2009@gxu.edu.cn.
FAU - Qu, Sujie
AU  - Qu S
AD  - Guangxi Center for Animal Disease Control and Prevention, Nanning, 530001, China.
FAU - Long, Feng
AU  - Long F
AD  - Guangxi Center for Animal Disease Control and Prevention, Nanning, 530001, China.
FAU - Lu, Wenjun
AU  - Lu W
AD  - Guangxi Center for Animal Disease Control and Prevention, Nanning, 530001, China.
LA  - eng
PT  - Journal Article
DEP - 20220118
PL  - England
TA  - BMC Vet Res
JT  - BMC veterinary research
JID - 101249759
SB  - IM
MH  - *African Swine Fever Virus/genetics
MH  - Animals
MH  - China/epidemiology
MH  - *Classical Swine Fever/diagnosis
MH  - *Classical Swine Fever Virus/genetics
MH  - *Pestivirus/genetics
MH  - Phylogeny
MH  - *Reverse Transcriptase Polymerase Chain Reaction/veterinary
MH  - Sensitivity and Specificity
MH  - Swine
MH  - *Swine Diseases/diagnosis
PMC - PMC8764768
COIS- The authors declare that they have no competing interests.
EDAT- 2022/01/20 06:00
MHDA- 2022/03/04 06:00
PMCR- 2022/01/18
CRDT- 2022/01/19 05:48
PHST- 2021/05/05 00:00 [received]
PHST- 2022/01/06 00:00 [accepted]
PHST- 2022/01/19 05:48 [entrez]
PHST- 2022/01/20 06:00 [pubmed]
PHST- 2022/03/04 06:00 [medline]
PHST- 2022/01/18 00:00 [pmc-release]
AID - 10.1186/s12917-022-03144-4 [pii]
AID - 3144 [pii]
AID - 10.1186/s12917-022-03144-4 [doi]
PST - epublish
SO  - BMC Vet Res. 2022 Jan 18;18(1):43. doi: 10.1186/s12917-022-03144-4.