[Development of one step RT-PCR technique for detection of H7 subtype avian influenza].
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.
Bao
Hongmei
H
State Key Laboratory of Veterinary Biotechnology, Animal Influenza Laboratory of Ministry of Agriculture, Harbin Veterinary Research Institute of CAAS, Harbin 150001, China.
Wang
Xiurong
X
Tao
Qimeng
Q
Cai
Dongdong
D
Wang
Fumei
F
Chen
Hualan
H
D000818:Animals; D001717:Birds; D002642:Chick Embryo; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics*; D057916:Influenza A Virus, H7N1 Subtype / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D019267:Hemagglutinin Glycoproteins, Influenza Virus; C109963:hemagglutinin, avian influenza A virus