Differential diagnosis of fish pathogenic rhabdoviruses by reverse transcriptase-dependent polymerase chain reaction.
Reverse transcriptase-dependent polymerase chain reaction (RT-PCR) was applied to the detection and differentiation of viral hemorrhagic septicemia virus (VHSV) and infectious hematopoietic necrosis virus (IHNV) using primer pairs designed for the amplification of glycoprotein G-specific gene fragments of the two viruses. The products of 443 bp (VHS) and 548 bp (IHN), respectively, were amplified from the total RNA extracts of RTG-2 cells infected with a total of 9 different strains of either VHS virus or IHN virus. Restriction analysis using FokI, and DNA sequencing of the PCR products demonstrated specificity of the amplification. The RT-PCR amplification of VHSV or IHNV G-genes was found to be a simple, highly specific and sensitive method allowing differential diagnosis of VHS and IHN within 8 h.
Bruchhof
B
B
Federal Research Centre for Virus Diseases of Animals, Tübingen, Germany.
Marquardt
O
O
Enzmann
P J
PJ
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D018076:DNA, Complementary; D004279:DNA, Viral / Q000032:analysis; D005069:Evaluation Studies as Topic; D005393:Fish Diseases / Q000175:diagnosis / Q000821:virology*; D008562:Membrane Glycoproteins / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis; D012194:RNA-Directed DNA Polymerase; D012209:Rhabdoviridae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D018353:Rhabdoviridae Infections / Q000097:blood / Q000662:veterinary* / Q000821:virology; D012680:Sensitivity and Specificity; D014337:Trout / Q000097:blood / Q000821:virology*; D014759:Viral Envelope Proteins / Q000235:genetics
D018076:DNA, Complementary; D004279:DNA, Viral; C096089:G protein, viral hemorrhagic septicemia virus; D008562:Membrane Glycoproteins; D012367:RNA, Viral; D014759:Viral Envelope Proteins; D012194:RNA-Directed DNA Polymerase