METHOD FOR THE MULTIPLE DETECTION OF INFLUENZA VIRUS TYPE A, TYPE B OR TYPE C BY MEANS OF OLIGONUCLEOTIDES DESIGNED TO ALLOW ALLELIC DISCRIMINATION IN A MULTIPLEX RT-PCR.

摘要

The invention relates to a broad-spectrum detection system having a specificity of 99% and being based on mechanisms for the multiple allelic differentiation of influenza virus type A (sub-types H1N1, H3N2, H5N1, H7N3), type B or type C. The main components of the system are sense oligonucleotides that allow allelic discrimination, said oligonucleotides exhibiting various characteristics including: a modification in the penultimate nucleotide of region 3' in order to generate a mismatch when hybridising to the target sequence of the M1 gene; the corresponding polymorphism of the sequence to be analysed being located in the last nucleotide of region 3'; and said oligonucleotides being specific for highly conserved regions having a low incidence of genetic drift. The system also comprises antisense oligonucleotides designed such that they are specific for non-conserved regions in order to guarantee their specificity for the specific region of the target sequence to be analysed. All the aforementioned forms part of one single reaction allowing the multiple differential analysis of the different types and sub-types of influenza, and leading to a reduction in the time it takes to obtain results and an increase in the viability thereof for use in epidemiological population studies.