title: Rapid detection of hog cholera virus in tissues by the polymerase chain reaction.
abstract: A rapid method for the detection of hog cholera virus (HCV) in infected tissues, using polymerase chain reaction (PCR) was developed. Total RNA isolated from HCV-infected tissues was reverse transcribed with AMV reverse transcriptase and the resulting complementary DNA was amplified by Taq DNA polymerase in the presence of two HCV-specific primers. The amplified DNA fragment was detected by agarose gel electrophoresis. The sensitivity of this method was at 10(4) TCID50 of HCV. The sensitivity increased approximately 1000-fold when the DNA was reamplified with a set of nested primers. DNA sequencing analysis of the PCR products revealed that the HCV sequence amplified from a local field isolate was highly homologous to the HCV Alfort strain. This method may be useful for pathological and epidemiological studies of HCV in pigs.
authors.affiliation: Department of Microbiology and Immunology, Chang-Gung Medical College, Taoyuan, Taiwan, R.O.C.
authors.forename: S T
authors.initials: ST
authors.lastname: Liu
authors.forename: S N
authors.initials: SN
authors.lastname: Li
authors.forename: D C
authors.initials: DC
authors.lastname: Wang
authors.forename: S F
authors.initials: SF
authors.lastname: Chang
authors.forename: S C
authors.initials: SC
authors.lastname: Chiang
authors.forename: W C
authors.initials: WC
authors.lastname: Ho
authors.forename: Y S
authors.initials: YS
authors.lastname: Chang
authors.forename: S S
authors.initials: SS
authors.lastname: Lai
mesh_terms: D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D006691:Classical Swine Fever / Q000175:diagnosis* / Q000235:genetics; D006692:Classical Swine Fever Virus / Q000235:genetics / Q000302:isolation & purification*; D003001:Cloning, Molecular; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012367:RNA, Viral / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013552:Swine
chemical_list: D004277:DNA, Single-Stranded; D004279:DNA, Viral; D012367:RNA, Viral