The invention belongs to the
biotechnology application field, and relates to simultaneous detection and
genotyping of infections of 16 respiratory viruses (including FluA, FluB, sH1N1, PIV1, PIV2, PIV3, RSVA, RSVB, HRV, HMPV, HBoV, CoV NL63, CoV OC43, CoV 229E, CoV HKU1 and Adv) of nasopharyngeal extract specimens of patients of respiratory-related diseases of all levels of
disease prevention and control institutions and sentinel hospitals. Specifically,
nucleotide sequences of representative strains of the 16 respiratory viruses are downloaded from NCBI; through literature review and
multiple sequence alignment,
pathogen relatively-conservative regions are determined, and
multiplex specific primers are designed. Single-tube
multiplex (18 stages) PCR detection is carried out for detecting the 16
respiratory virus conservative regions, and an entire reaction takes less than 2 hours. According to the invention, a defect that
genotyping cannot be carried out with conventional single-tube
multiplex fluorescent qualitative PCR detections can be overcome, and defects of complicated operations, long time, and high cost of conventional
chip detection methods can be overcome. With the application provided by the invention, a novel idea is provided for
respiratory virus genotyping technologies. With characteristics of high specificity, high sensitivity, and high speed, powerful technological support is provided for rapid and accurate screening and genotyping of the respiratory-
disease-related viruses. The invention has important significance upon the researches of respiratory-tract patient infection
pathogen spectrum of out nation, and upon molecular epidemiological investigations.