title: Development of an updated PCR assay for detection of African swine fever virus.
abstract: Due to the current unavailability of vaccines or treatments for African swine fever (ASF), which is caused by African swine fever virus (ASFV), rapid and reliable detection of the virus is essential for timely implementation of emergency control measures and differentiation of ASF from other swine diseases with similar clinical presentations. Here, an improved PCR assay was developed and evaluated for sensitive and universal detection of ASFV. Primers specific for ASFV were designed based on the highly conserved region of the vp72 gene sequences of all ASFV strains available in GenBank, and the PCR assay was established and compared with two OIE-validated PCR tests. The analytic detection limit of the PCR assay was 60 DNA copies per reaction. No amplification signal was observed for several other porcine viruses. The novel PCR assay was more sensitive than two OIE-validated PCR assays when testing 14 strains of ASFV representing four genotypes (I, V, VIII and IX) from diverse geographical areas. A total of 62 clinical swine blood samples collected from Uganda were examined by the novel PCR, giving a high agreement (59/62) with a superior sensitive universal probe library-based real-time PCR. Eight out of 62 samples tested positive, and three samples with higher Ct values (39.15, 38.39 and 37.41) in the real-time PCR were negative for ASFV in the novel PCR. In contrast, one (with a Ct value of 29.75 by the real-time PCR) and two (with Ct values of 29.75 and 33.12) ASFV-positive samples were not identified by the two OIE-validated PCR assays, respectively. Taken together, these data show that the novel PCR assay is specific, sensitive, and applicable for molecular diagnosis and surveillance of ASF.
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute (HVRI), Chinese Academy of Agricultural Sciences, 678 Haping Road, Harbin, 150069, China.|Joint Laboratory on Veterinary Microbiology between HVRI and SVA, 678 Haping Road, Harbin, 150069, China.
authors.forename: Yuzi
authors.initials: Y
authors.lastname: Luo
authors.affiliation: National Animal Disease Diagnostics and Epidemiology Center (NADDEC), Ministry of Agricultural, Animal Industry and Fisheries, Entebbe, Uganda.
authors.forename: Stella A
authors.initials: SA
authors.lastname: Atim
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute (HVRI), Chinese Academy of Agricultural Sciences, 678 Haping Road, Harbin, 150069, China.|Joint Laboratory on Veterinary Microbiology between HVRI and SVA, 678 Haping Road, Harbin, 150069, China.
authors.forename: Lina
authors.initials: L
authors.lastname: Shao
authors.affiliation: National Animal Disease Diagnostics and Epidemiology Center (NADDEC), Ministry of Agricultural, Animal Industry and Fisheries, Entebbe, Uganda.
authors.forename: Chrisostom
authors.initials: C
authors.lastname: Ayebazibwe
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute (HVRI), Chinese Academy of Agricultural Sciences, 678 Haping Road, Harbin, 150069, China.|Joint Laboratory on Veterinary Microbiology between HVRI and SVA, 678 Haping Road, Harbin, 150069, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Sun
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute (HVRI), Chinese Academy of Agricultural Sciences, 678 Haping Road, Harbin, 150069, China.|Joint Laboratory on Veterinary Microbiology between HVRI and SVA, 678 Haping Road, Harbin, 150069, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute (HVRI), Chinese Academy of Agricultural Sciences, 678 Haping Road, Harbin, 150069, China.|Joint Laboratory on Veterinary Microbiology between HVRI and SVA, 678 Haping Road, Harbin, 150069, China.
authors.forename: Shengwei
authors.initials: S
authors.lastname: Ji
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute (HVRI), Chinese Academy of Agricultural Sciences, 678 Haping Road, Harbin, 150069, China.|Joint Laboratory on Veterinary Microbiology between HVRI and SVA, 678 Haping Road, Harbin, 150069, China.
authors.forename: Xing-Yu
authors.initials: XY
authors.lastname: Meng
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute (HVRI), Chinese Academy of Agricultural Sciences, 678 Haping Road, Harbin, 150069, China.|Joint Laboratory on Veterinary Microbiology between HVRI and SVA, 678 Haping Road, Harbin, 150069, China.
authors.forename: Su
authors.initials: S
authors.lastname: Li
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute (HVRI), Chinese Academy of Agricultural Sciences, 678 Haping Road, Harbin, 150069, China.|Joint Laboratory on Veterinary Microbiology between HVRI and SVA, 678 Haping Road, Harbin, 150069, China.
authors.forename: Yongfeng
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Biological Sciences, College of Natural Sciences, Makerere University, P. O. Box 7062, Kampala, Uganda.
authors.forename: Charles
authors.initials: C
authors.lastname: Masembe
authors.affiliation: Department for Disease Control and Epidemiology, National Veterinary Institute (SVA), 75189, Uppsala, Sweden.|Joint Laboratory on Veterinary Microbiology between HVRI and SVA, 678 Haping Road, Harbin, 150069, China.
authors.forename: Karl
authors.initials: K
authors.lastname: Ståhl
authors.affiliation: Department of Microbiology, National Veterinary Institute (SVA), 75189, Uppsala, Sweden.|Joint Laboratory on Veterinary Microbiology between HVRI and SVA, 678 Haping Road, Harbin, 150069, China.
authors.forename: Frederik
authors.initials: F
authors.lastname: Widén
authors.affiliation: Department of Microbiology, National Veterinary Institute (SVA), 75189, Uppsala, Sweden.|Joint Laboratory on Veterinary Microbiology between HVRI and SVA, 678 Haping Road, Harbin, 150069, China.
authors.forename: Lihong
authors.initials: L
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute (HVRI), Chinese Academy of Agricultural Sciences, 678 Haping Road, Harbin, 150069, China. qiuhuaji@163.com.|Joint Laboratory on Veterinary Microbiology between HVRI and SVA, 678 Haping Road, Harbin, 150069, China. qiuhuaji@163.com.
authors.forename: Hua-Ji
authors.initials: HJ
authors.lastname: Qiu
mesh_terms: D000357:African Swine Fever / Q000175:diagnosis*; D000358:African Swine Fever Virus / Q000235:genetics / Q000302:isolation & purification*; D000818:Animals; D017931:DNA Primers / Q000235:genetics; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013552:Swine; D014454:Uganda
chemical_list: D017931:DNA Primers