Summary
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.
References
Bentley DR, Brownlee GG (1982) Sequence of the N2 neuraminidase from influenza virus A/NT/60/68. Nucleic Acids Res 10: 5033–5042
Bressoud A, Whitcomb J, Pourzand C, Haller O, Cerutti P (1990) Rapid detection of influenza virus H1 by the polymerase chain reaction. Biochem Biophys Res Commun 167: 425–430
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159
Chomel JJ, Thouvenot D, Onno M, Kaiser C, Gourreau JM, Aymard M (1989) Rapid diagnosis of influenza infection of NP antigen using an immunocapture ELISA test. J Virol Methods 25: 81–92
Claas ECJ, Sprenger MJW, Kleter GEM, van Beek R, Quint WGV, Masurel N (1992) Type-specific identification of influenza viruses A, B and C by the polymerase chain reaction. J Virol Methods 39: 1–13
Concannon P, Kwolek CJ, Salser WA (1984) Nucleotide sequence of the influenza virus A/USSR/90/77 neuraminidase gene. J Virol 50: 654–656
Coonrod JD, Karathanasis P, Betts RF, Donofrio JC (1988) Enzyme-linked immunosorbent assay of core antigens for clinical diagnosis of influenza. J Med Virol 25: 399–409
Donofrio JC, Coonrod D, Davidson JN, Betts RF (1992) Detection of influenza A and B in respiratory secretions with the polymerase chain reaction. PCR Methods Appl 1: 263–280
Elleman TC, Azad AA, Ward CW (1982) Neuraminidase gene from the early Asian strain of human influenza A virus A/RI/5−/57 (H2N2). Nucleic Acids Res 10: 7005–7015
Hiti AL, Nayak DP (1982) Complete nucleotide sequence of the neuraminidase gene of human influenza virus A/WSN/33. J Virol 41: 730–734
Lentz MR, Air GM, Laver WG, Webster RG (1984) Sequence of the neuraminidase gene of influenza virus A/Tokyo/3/67 and previously uncharacterized monoclonal variants. Virology 135: 257–265
Mantero G, Zonaro A, Albertini A, Bertolo P, Primi D (1991) DNA enzyme immunoassay: general method for detecting products of polymerase chain reaction. Clin Chem 37: 422–429
Markoff L, Lai CJ (1982) Sequence of the influenza A/Udorn/72 (H3N2) virus neuraminidase gene as determined from cloned full-length DNA. Virology 119: 288–297
Martìnez C, Del Rio L, Portela A, Domingo E, Ortìn J (1983) Evolution of the influenza virus neuraminidase gene during drift of the N2 subtype. Virology 130: 539–545
Pisareva M, Bechtereva T, Plyusnin A, Dobretsova A, Kisselev O (1992) PCR-amplification of influenza A virus specific sequences. Arch Virol 5: 313–331
Schreier E, Roeske H, Driesel G, Künkel U, Petzold DR, Berlinghoff R, Michel S (1988) Complete nucleotide sequence of the neuraminidase gene of the human influenza virus A/Chile/1/83 (H1N1). Arch Virol 99: 271–276
Schreier E, Schweiger B, Ramelow C, Beziat P, Süss J (1994) Rapid detection of tick-borne encephalitis virus sequences by cDNA amplification coupled to a simple DNA enzyme immunoassay. Clin Diagn Virol 2: 291–295
Stokes CE, Bernstein JM, Kyger SA, Hayden FG (1988) Rapid diagnosis of influenza A and B by 24-h fluorescent focus assay. J Clin Microbiol 26: 1263–1266
Van Rompuy L, Min Jou W, Huylebroeck D, Fiers W (1982) Complete nucleotide sequence of a human influenza neuraminidase gene of subtype N2 (A/Victoria/3/75). J Mol Biol 161: 1–11
Viazov S, Zibert A, Widell A, Cavicchini A, Schreier E, Roggendorf M (1994) Typing of hepatitis C virus isolates by DNA enzyme immunoassay. J Virol Methods 48: 81–92
Yamada A, Imanishi J (1992) Detection of influenza B virus in throat swabs using the polymerase chain reaction. Acta Virol 36: 320–325
Zhang W, Evans DH (1991) Detection and identification of human influenza viruses by the polymerase chain reaction. J Virol Methods 33: 165–189
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Schweiger, B., Lange, I., Heckler, R. et al. Rapid detection of influenza A neuraminidase subtypes by cDNA amplification coupled to a simple DNA enzyme immunoassay. Archives of Virology 139, 439–444 (1994). https://doi.org/10.1007/BF01310805
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01310805