Differentiation of infectious pancreatic necrosis virus isolates by polymerase chain reaction.
Infectious pancreatic necrosis virus (IPNV) from lake trout of Cornwall lake (LT-IPNV), from trout of Jasper (Ja-IPNV) and Arctic char of Northwest Territories (AC-IPNV) are the three isolates recorded from western Canada. Two segments (nt 453-674 and nt 1197-1503) from VP2 coding region of RNA of the isolates were amplified by polymerase chain reaction (PCR) for differentiation. Primers for cDNA synthesis and amplification by PCR were chosen from VP2 coding region of RNA of Ja-IPNV. The segment of 453-674 could be amplified in all the three isolates at annealing temperature from 50 to 60 degrees C. However the segment nt 1197-1503 could be amplified only from Jasper and not from AC or LT-INV at primer annealing temperature ranging from 50 degrees-60 degrees C. PCR product could be obtained from the latter two isolates only at Primer annealing temperature of 45 degrees C. This difference in annealing temperature in PCR amplification between the isolates could be used for differentiating the isolates at genome level.
Shankar
K M
KM
Department of Microbiology, University of Alberta, Edmonton, Canada.
Roy
K L
KL
Yamamoto
T
T
D001483:Base Sequence; D003937:Diagnosis, Differential; D018061:Infectious pancreatic necrosis virus / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction