The present application relates to the technical field of biological detection, and in particular to a detection primer and
nucleic acid mass spectrometry detection method for African horse sickness
virus and
West Nile virus. Based on the MALDI-TOF
nucleic acid mass spectrometry detection method, the present application selects two relatively conservative genes, VP7 and NS2, as target genes for African horse sickness
virus; and for
West Nile virus, two equally conservative genes, NS2A and NS5, are selected. Based on these target genes, the present application designs four pairs of amplification primers and four extension primers. Through optimization of the selection of amplification sites and addition of modified bases, the molecular weight of the extension probe is effectively distinguished within the detection range, thereby achieving the goal of
multiplex detection, and at the same time ensuring high accuracy, specificity and sensitivity of the detection. The present application can not only efficiently, sensitively and accurately identify the infection of African horse sickness
virus and
West Nile virus, but also can be flexibly adjusted and optimized according to actual needs, and is expected to play an important role in future
virus detection and
differential diagnosis.