1.Development of one step RT-PCR technique for detection of H7 subtype avian influenza.
Hongmei BAO ; Xiurong WANG ; Qimeng TAO ; Dongdong CAI ; Fumei WANG ; Hualan CHEN
Chinese Journal of Biotechnology 2009;25(11):1658-1663
According to 45 hemagglutinin (HA) gene sequences of H7 subtype of avian influenza virus (AIV), a pair of specific oligonucleotide primers was designed. We developed one step RT-PCR for detecting AIV subtype H7. Sensitivity to detection of allantoic fluid by one step RT-PCR reached 10(5.5) EID50/mL and detection of swab samples reached 10(3) EID50/mL. We simultaneity detected the tissue and swab samples infected with H7 subtypes AIV by one step RT-PCR and virus isolation method. The results showed that the sensitivity of the assay gave an excellent correlation with the conventional virus isolation method. H1-H15 subtypes of avian influenza and other avian diseases were detected by the one step RT-PCR. The results showed the assays were high specific, without cross-reaction with other subtypes or other avian diseases. Development of one step RT-PCR will provide effective technical support for the rapid diagnosis and surveillance of molecular epidemiology of AIV subtype H7.
Animals
;
Birds
;
Chick Embryo
;
Hemagglutinin Glycoproteins, Influenza Virus
;
genetics
;
Influenza A Virus, H7N1 Subtype
;
classification
;
genetics
;
isolation & purification
;
Influenza in Birds
;
virology
;
Reverse Transcriptase Polymerase Chain Reaction
;
methods
;
Sensitivity and Specificity