Application of real-time RT-PCR utilising lux (light upon extension) fluorogenic primer for the rapid detection of avian influenza viruses.
A real-time RT-PCR assay utilising light upon extension fluorogenic primer (LUX RT-PCR) was developed for the rapid and efficient detection of avian influenza viruses (AIV). The assay detected each of the AIV isolates tested (16/16) and gave negative results with heterologous pathogens (17/17). The detection limit of the assay proved to be 10(-0.5) EID50/0.2 ml and 10(1.5) EID50/0.2 ml in allantoic fluid of virus-infected embryonated chicken eggs and in spiked chicken faeces samples, respectively. Based on its specificity, sensitivity and relative simplicity, the LUX RT-PCR assay provides a novel, rapid and cost-effective diagnostic tool for avian influenza surveillance and monitoring programs.
Kiss
I
I
Central Veterinary Institute, Institute of Debrecen, H-4031 Debrecen, Bornemissza u. 3-7, Hungary. kissi@oai.hu
Germán
P
P
Sámi
L
L
Antal
Márta
M
Farkas
T
T
Kardos
G
G
Kecskeméti
S
S
Dán
A
A
Belák
S
S
D000818:Animals; D001715:Bird Diseases / Q000175:diagnosis* / Q000821:virology; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis*; D005453:Fluorescence; D005585:Influenza in Birds / Q000175:diagnosis* / Q000821:virology; D009975:Orthomyxoviridae / Q000235:genetics*; D011200:Poultry; D011237:Predictive Value of Tests; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004279:DNA, Viral