The invention relates to the technical field of
gene engineering, in particular to a circulating in-situ hybridization method for detecting
RNA (Ribonucleic Acid) in tissue. According to the invention, a microfluidic
system is not needed, the
cover glass is treated and coated, the tissue slice is attached to the coated
cover glass, the probe hybridization
pool is attached to the periphery of the tissue, and a closed device for
hybridization reaction is provided. The method comprises the following steps: carrying out in-situ hybridization on
RNA sites by using lock-type probes, connecting the lock-type probes hybridized to correct sites into a ring through ligase, amplifying the lock-type probes connected into the ring by using a rolling circle amplification technology, converting an
RNA signal into a single-stranded
DNA signal and simultaneously completing
signal amplification, and enabling each lock-type probe to contain a unique
barcode base sequence, the method is used for combining with a fluorescent probe to detect RNA of a corresponding bar code sequence. The method is high in
signal intensity, collection of
fluorescence signal points does not depend on a super-resolution imaging platform and a microfluidic
system, and multi-
gene in-situ detection which is low in cost and easy to operate can be achieved.