Differentiation between Cyprinid herpesvirus type-3 lineages using duplex PCR.
To date, all the isolates of Cyprinid herpesvirus type-3 (CyHV3) responsible for serious outbreaks in carps Cyprinus carpio have been found to be very similar or identical on the basis of DNA sequences of a few reference genes. However, two genetic lineages (U/I and J) are distinguished by full-length genome sequencing. Two molecular markers presenting genetic variations were targeted for developing a duplex PCR assay able to distinguish CyHV3-U/I from CyHV3-J while avoiding DNA sequencing. The method was validated on a series of 42 samples of infected carps from France, The Netherlands and Poland collected from 2001 to 2008. Among these samples, both the U/I and J genotypes were identified, but also a third genotype representing a genetic intermediate between U/I and J for one of the two molecular markers. A classification of CyHV3 genotypes, based on the alleles of the two molecular markers, is proposed. The assay is easy to perform and provides a genotype information with samples moderately or highly concentrated. This tool should improve our knowledge regarding the present distribution and future diversification of this emerging virus.
Bigarré
L
L
Pathologie Virale des Poissons, AFSSA, Technopôle Brest-Iroise, Plouzané, France. l.bigarre@afssa.fr
Baud
M
M
Cabon
J
J
Antychowicz
J
J
Bergmann
S M
SM
Engelsma
M
M
Pozet
F
F
Reichert
M
M
Castric
J
J
D000818:Animals; D002347:Carps / Q000821:virology*; D017931:DNA Primers / Q000235:genetics; D005602:France; D005838:Genotype; D006564:Herpesviridae / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D006566:Herpesviridae Infections / Q000821:virology*; D009426:Netherlands; D011044:Poland; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers