The invention discloses a panda-derived feline panleucopenia
virus RAA-CRISPRCas12a detection method, and belongs to the technical field of
virus detection, and the panda-derived feline panleucopenia
virus RAA-CRISPRCas12a detection method comprises the following steps: S1, primer and probe design; s2, screening an RAA amplification primer; s3, RAA reaction condition optimization; s4, performing RAA sensitivity and specificity
verification; s5, preparing crRNA (Complementary Ribonucleic Acid); s6, optimal crRNA screening is carried out; s7, optimizing the RAA-
CRISPR / Cas12a (Reverse Amplified Architecture- S8, the specificity and the sensitivity of the RAA-
CRISPR / Cas12a are verified; s9, clinical sample detection; according to the invention, by elaborately designing a primer and a probe aiming at the
capsid protein VP2
gene of the
feline parvovirus (FPV), fully utilizing the advantages of
recombinase-mediated amplification (RAA) and a
CRISPR / Cas12a
system, and comprehensively optimizing
reaction conditions and components of the
system, the detection method has remarkable advantages, can quickly complete the detection process under the condition of constant temperature of 37 DEG C, is extremely high in sensitivity, and can be used for rapidly detecting the
feline parvovirus. The lowest
detection limit can reach 1 * 10 < 1 > copies / mu L, the specificity is high, and the
coincidence rate with a real-time fluorescent quantitative
PCR method is as high as 100%.