Development of one step RT-PCR technique for detection of H7 subtype avian influenza.
- Author:
Hongmei BAO
1
;
Xiurong WANG
;
Qimeng TAO
;
Dongdong CAI
;
Fumei WANG
;
Hualan CHEN
Author Information
1. State Key Laboratory of Veterinary Biotechnology, Animal Influenza Laboratory of Ministry of Agriculture, Harbin Veterinary Research Institute of CAAS, Harbin 150001, China.
- Publication Type:Journal Article
- MeSH:
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
- From:
Chinese Journal of Biotechnology
2009;25(11):1658-1663
- CountryChina
- Language:Chinese
-
Abstract:
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.
