Detection of Cyprinid herpesvirus-3 (CyHV-3) DNA in infected fish tissues by nested polymerase chain reaction.
Diagnostic tests with high analytical sensitivity are required to detect Cyprinid herpesvirus-3 (CyHV-3) in carriers that have been implicated in the dissemination of this important disease of koi and common carp Cyprinus carpio. A nested polymerase chain reaction (PCR) test was developed to detect CyHV-3 DNA in tissues of infected fish. The 2-round PCR amplified 529 and 379 bp segments of the CyHV-3 gene coding for the major capsid protein using specific external and internal primers, respectively. The PCR test did not amplify genomic DNA from either Cyprinid herpesvirus-1 (CyHV-1) or Cyprinid herpesvirus-2 (CyHV-2). The nested PCR test detected CyHV-3 DNA at a 10-fold lower concentration than a routinely used, 1-round test. Hence, the new method should provide more sensitive diagnosis of CyHV-3 infection among koi and carp populations.
El-Matbouli
Mansour
M
Institute of Zoology, Fish Biology and Fish Diseases, Faculty of Veterinary Medicine, University of Munich, Germany. el-matbouli@lmu.de
Rucker
Ute
U
Soliman
Hatem
H
D000818:Animals; D001921:Brain / Q000821:virology; D036022:Capsid Proteins / Q000235:genetics; D002347:Carps; D004279:DNA, Viral / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D005393:Fish Diseases / Q000175:diagnosis / Q000821:virology*; D005880:Gills / Q000821:virology; D006564:Herpesviridae / Q000235:genetics* / Q000302:isolation & purification; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D007668:Kidney / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013154:Spleen / Q000821:virology
D036022:Capsid Proteins; D004279:DNA, Viral