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The invention discloses a kit for synchronously detecting twenty-two respiratory tract pathogens and a detection method of the kit. The kit comprises DEPC (diethylpyrocarbonate) water, a 5*RT (reverse transcription) buffer, a reverse transcription primer, a reverse transcriptase, an X solution, a 10*PCR (polymerase chain reaction) buffer, a PCR primer, a 25mM magnesium chloride solution, a DNA (deoxyribonucleic acid) polymerase and a positive control, and is characterized in that the reverse transcription primer comprises RT amplification primers of the nine respiratory tract pathogens, and a reverse primer of a human RNA (ribonucleic acid) internal reference, and has a gene sequence shown as SEQ ID NO. 1-6 (sequence identifier number 1-6), and the PCR primer comprises forward and reverse PCR amplification primers of the rest thirteen respiratory tract pathogens, a forward and reverse amplification primer of a human DNA internal reference, a forward and reverse PCR amplification primer of a reaction internal reference, PCR amplification primers of the nine respiratory tract pathogens, and a PCR amplification primer of the human RNA internal reference, and has a gene sequence shown as SEQ ID NO. 7-44. The kit and the detection method have the advantages of high specificity, sensitivity, flux and reliability, low cost, and no false negative results.