Rapid and sensitive detection of avian influenza virus subtype H7 using NASBA.
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.
Collins
Richard A
RA
Hong Kong DNA Chips Ltd., 1805-6, 18/F, Lu Plaza, 2 Wing Yip Street, Kowloon, Hong Kong SAR, China.
Ko
Lung-Sang
LS
Fung
King-Yip
KY
Chan
Ka-Yun
KY
Xing
Jun
J
Lau
Lok-Ting
LT
Yu
Albert Cheung Hoi
AC
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
C034477:RNA primers; D012367:RNA, Viral; D012313:RNA; D011189:Potassium Chloride