Rapid detection of hog cholera virus in tissues by the polymerase chain reaction.
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.
Liu
S T
ST
Department of Microbiology and Immunology, Chang-Gung Medical College, Taoyuan, Taiwan, R.O.C.
Li
S N
SN
Wang
D C
DC
Chang
S F
SF
Chiang
S C
SC
Ho
W C
WC
Chang
Y S
YS
Lai
S S
SS
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
D004277:DNA, Single-Stranded; D004279:DNA, Viral; D012367:RNA, Viral