The invention discloses a kit for detecting B and E type human adenovirus nucleic acid and a detection method thereof. The kit for detecting the B and E type human adenovirus nucleic acid comprises RPA freeze-dried particles, an RPA reaction primer, an RPA reaction probe, an RPA buffer solution, a magnesium salt solution, a transverse flow test strip, a buffer solution, double distilled water and an adenovirus genome DNA. The kit for detecting the B and E type human adenovirus nucleic acid has the advantages that the transverse flow test strip is utilized for carrying out visual qualitative detection on an RPA nucleic acid amplification product, the whole reaction is carried out at the constant temperature of 38 DEG C, a stripe on the test strip can be observed by virtue of naked eyes within 5-20 minutes, results are read and discriminated, and a special detecting instrument is not needed during detection. The kit disclosed by the invention has the characteristics of high sensitivity, strong specificity, easiness in operation, quick reaction and portability of instruments and is especially applicable to bedside rapid diagnosis of adenovirus, epidemic prevention and disease control.