title: Rapid and sensitive detection of avian influenza virus subtype H7 using NASBA.
abstract: Nucleic acid sequence-based amplification with electrochemiluminescent detection (NASBA/ECL) is an isothermal technique allowing rapid amplification and detection of specific regions of nucleic acid from a diverse range of sources. It is especially suitable for amplifying RNA. A NASBA/ECL technique has been developed allowing the detection of RNA from avian influenza virus subtype H7 derived from allantoic fluid harvested from inoculated chick embryos and from cell cultures. Degenerate amplification primers and amplicon capture probes were designed enabling the detection of low and highly pathogenic avian influenza of the H7 subtype from the Eurasian and North American lineages and the Australian sub-lineage. The NASBA/ECL technique is specific for subtype H7 and does not cross-react with other influenza subtypes or with viruses containing haemagglutinin-like genes. The assay is 10- to 100-fold more sensitive than a commercially available antigen capture immunoassay system. The NASBA/ECL assay could be used in high throughput poultry screening programmes.
authors.affiliation: Hong Kong DNA Chips Ltd., 1805-6, 18/F, Lu Plaza, 2 Wing Yip Street, Kowloon, Hong Kong SAR, China.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Collins
authors.forename: Lung-Sang
authors.initials: LS
authors.lastname: Ko
authors.forename: King-Yip
authors.initials: KY
authors.lastname: Fung
authors.forename: Ka-Yun
authors.initials: KY
authors.lastname: Chan
authors.forename: Jun
authors.initials: J
authors.lastname: Xing
authors.forename: Lok-Ting
authors.initials: LT
authors.lastname: Lau
authors.forename: Albert Cheung Hoi
authors.initials: AC
authors.lastname: Yu
mesh_terms: D000818:Animals; D001483:Base Sequence; D002642:Chick Embryo; D004563:Electrochemistry / Q000379:methods; D009980:Influenza A virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D008163:Luminescent Measurements; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D011189:Potassium Chloride / Q000494:pharmacology; D011201:Poultry Diseases / Q000821:virology; D012313:RNA; D012367:RNA, Viral / Q000032:analysis*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D013997:Time Factors
chemical_list: C034477:RNA primers; D012367:RNA, Viral; D012313:RNA; D011189:Potassium Chloride