Establishment and application of PCV-2, PCV-3 and PCV-4 triple PCR (Polymerase Chain Reaction) identification and detection method

• Main Authors: OU YUNWEN, WANG QIN, LIU LIJUN
• Format: Patent
• Language: Chinese; English
• Published: 13.05.2022
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention discloses a primer and a method for triple PCR (Polymerase Chain Reaction) identification and detection of PCV-2, PCV-3 and PCV-4 and application of the primer and the method in preparation of a triple PCR identification and detection kit. The primer of the PCV-2 is P1: TGTTTTCGAAGGCA, P2: ACAGCCCTAACCTATG, and the primer of the PCV-4 is a primer of the PCV-4. The primer of the PCV-3 is P3: GAGCGCTCGTAGTTCC, and the primer of the PCV-3 is P4: AGAAGGCGCTATGTC; the primer of the PCV-4 is P5: TGTTTGGGGTAGTTA, and the primer of the PCV-4 is P6: CAATCAGATCTAGG. The reaction system comprises 25.0 [mu] L of 2 * Taq MasterMix, 0.5 [mu] L of primers, 3.0 [mu] L of a template and 19.0 [mu] L of H2O; the reaction procedure is that the temperature is 95 DEG C for 5 minutes; the temperature is 95 DEG C for 40 seconds, the temperature is 50 DEG C for 40 seconds, the temperature is 72 DEG C for 40 seconds, and 30 cycles are carried out; the temperature is 72 DEG C for 10 minutes. The established PCV-2, PCV-3 a