Rapid detection of influenza A neuraminidase subtypes by cDNA amplification coupled to a simple DNA enzyme immunoassay.
A newly developed colorimetric method, DNA enzyme immunoassay (DEIA), was applied to the detection of neuraminidase subtypes N1 and N2 of influenza A viruses. Reverse transcription and polymerase chain reaction with universal primers were used for genomic amplification of H1N1, H2N2, and H3N2 strains. Following amplification, an aliquot of the PCR product was hybridized to biotinylated DNA sequences (N1/N2 probes) immobilized on microtiter wells. The hybridization event was revealed by monoclonal antibodies to double stranded DNA in a standard ELISA reaction. The assay described here was able to distinguish accurately between the two neuraminidase subtypes of human influenza A viruses. It is a simple and rapid method facilitating the handling of a large number of samples and therefore seems to be easily applicable to diagnostic laboratories.
Schweiger
B
B
Robert Koch-Institut, Berlin, Federal Republic of Germany.
Lange
I
I
Heckler
R
R
Willers
H
H
Schreier
E
E
D001483:Base Sequence; D015342:DNA Probes; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics*; D016679:Genome, Viral; D007124:Immunoenzyme Techniques; D009980:Influenza A virus / Q000145:classification* / Q000201:enzymology / Q000235:genetics; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000145:classification* / Q000235:genetics; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D015342:DNA Probes; D018076:DNA, Complementary; D009439:Neuraminidase