The invention relates to the technical field of molecular biological detection of an
insect-borne infectious
disease, and discloses a triple real-time
fluorescence RT-PCR detection method, primer and probe as well as kit of
Zika virus,
dengue virus and
chikungunya virus. The primer comprises the sequences of SEQ ID No.1, SEQ ID No.2, SEQ ID No.4, SEQ ID No.5, SEQ ID No.7 and SEQ ID No.8; the probe comprises the sequences of SEQ ID No.3, SEQ ID No.6, and SEQ ID No.9; the kit comprises a primer probe
mixed solution, an RT-PCR reaction solution, an
enzyme mixed solution, and a positive standard substance. The
Zika virus, the
dengue virus and the
chikungunya virus can be fast detected in the reaction solution of a same tube by use of a triple real-time
fluorescence RT-PCR amplification technology by optimizing the reaction solution formula and the primer probe sequences, the single detection is unnecessary; compared with the traditional PCR detection method, the operation step is reduced, whether the reaction solution contains the three
virus can be fast and accurately detected, the detection time is shortened, the detection efficiency is improved, and the cost is saved.