title: A multiplex RT-PCR for rapid and simultaneous detection of porcine teschovirus, classical swine fever virus, and porcine reproductive and respiratory syndrome virus in clinical specimens.
abstract: A multiplex RT-PCR (mRT-PCR) assay was developed and evaluated for its ability to detect multiple viral infections of swine simultaneously. One pair of primers was selected carefully for each of the following three RNA viruses: porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV), and porcine teschovirus (PTV). Each target produced a specific amplicon with a size of 451bp (PRRSV), 343bp (CSFV), or 163bp (PTV). The sensitivity of the mRT-PCR using purified plasmid constructs containing the specific viral target fragments was 2.02 x 10², 2.90 x 10³, and 6.16 x 10³ copies for PRRSV, CSFV, and PTV, respectively. Among 69 clinical samples from Heilongjiang, Jilin, and Henan provinces, co-infection by PRRSV and CSFV was 4.4%, co-infection by PRRSV and PTV was 11.6%, co-infection by PTV and CSFV was 13.0%, and co-infection by the three viruses was 8.7%. In conclusion, the mRT-PCR should be useful for routine molecular diagnosis and epidemiology.
authors.affiliation: Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, No. 427, Maduan Street, Nangang District, Harbin 150001, China.
authors.forename: Shanshan
authors.initials: S
authors.lastname: Liu
authors.forename: Yarong
authors.initials: Y
authors.lastname: Zhao
authors.forename: Quanbo
authors.initials: Q
authors.lastname: Hu
authors.forename: Chaochao
authors.initials: C
authors.lastname: Lv
authors.forename: Chaofan
authors.initials: C
authors.lastname: Zhang
authors.forename: Rui
authors.initials: R
authors.lastname: Zhao
authors.forename: Feng
authors.initials: F
authors.lastname: Hu
authors.forename: Wencheng
authors.initials: W
authors.lastname: Lin
authors.forename: Shangjin
authors.initials: S
authors.lastname: Cui
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D006692:Classical Swine Fever Virus / Q000235:genetics / Q000502:physiology; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics / Q000502:physiology; D012327:RNA Virus Infections / Q000175:diagnosis / Q000662:veterinary*; D012328:RNA Viruses / Q000235:genetics / Q000502:physiology*; D012367:RNA, Viral; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology*; D052700:Teschovirus / Q000235:genetics / Q000502:physiology
chemical_list: D012367:RNA, Viral