RT-PCR-ELISA as a tool for diagnosis of low-pathogenicity avian influenza.
A one-tube reverse transcriptase/polymerase chain reaction coupled with an enzyme-linked immunosorbent assay (RT-PCR-ELISA) was developed for the rapid detection of avian influenza virus (AIV) in clinical specimens. A total of 419 swab pools were analyzed from chickens experimentally infected with low-pathogenicity AIV, from wild aquatic birds, and from domestic ducks. The AIV was detected in 32 swab pools by RT-PCR-ELISA compared to 23 by virus isolation (VI) in embryonated specific pathogen free (SPF) chicken eggs. Thus, 39% more specimens were positive by RT-PCR-ELISA than by VI. Two of the twenty-three VI-positive specimens were negative when tested by RT-PCR-ELISA. The diagnostic sensitivity and specificity of the RT-PCR-ELISA was 91% and 97%, respectively, using VI in SPF eggs as the gold reference standard.
Danish Veterinary Institute, Hangevej 2, DK-8200 Aarhus N, Denmark.
Karen
K
Dybkaer
Mette
M
Munch
Kurt J
KJ
Handberg
Poul H
PH
Jørgensen
D000482:Allantois / Q000821:virology; D000818:Animals; D001483:Base Sequence; D001717:Birds; D002642:Chick Embryo / Q000821:virology; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000145:classification / Q000276:immunology; D009980:Influenza A virus / Q000302:isolation & purification / Q000472:pathogenicity*; D005585:Influenza in Birds / Q000175:diagnosis* / Q000276:immunology; D016133:Polymerase Chain Reaction / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction; D021161:Self-Sustained Sequence Replication / Q000379:methods
D017931:DNA Primers; D019267:Hemagglutinin Glycoproteins, Influenza Virus