The invention belongs to the field of
biology, relates to a
salmonella gene detection technology, and discloses a one-pot detection method combining LAMP (loop-mediated isothermal amplification) and
CRISPR / BrCas12b mediated
nucleic acid molecule detection. The method comprises the following steps: designing an LAMP primer, a small
guide RNA (Ribonucleic Acid) and a transcription template overlapping PCR (
Polymerase Chain Reaction) primer, performing loop-mediated isothermal amplification, constructing
CRISPR / Cas shearing recognition (Cas-LAMP), constructing a single-tube one-pot
reaction system of the
CRISPR / Cas shearing recognition, detecting the specificity and sensitivity of Cas-LAMP amplification, and the like. In order to solve the technical problem that amplification and shearing recognition cannot be carried out synchronously, the invention provides a Cas-LAMP technology capable of linking two-step reaction of amplification and shearing recognition, and accurate, sensitive and
rapid detection of
nucleic acid molecules is realized in an in-vitro single
reaction tube system. Compared with a traditional
nucleic acid molecule detection method, the method has the following advantages that one-pot detection can be directly carried out without step-by-step operation, high sensitivity and strong specificity are achieved, and rapid
field detection can be achieved. The method provides
technical support for conventional
nucleic acid detection, and has a
wide field instant detection application prospect.