PCR (Polymerase Chain Reaction) method for amplifying N-terminal hypervariable region of S gene of porcine epidemic diarrhea virus

• Main Authors: ZHUANG XIAOLONG, GUO WEILU, FAN BAOCHAO, ZHU XUEJIAO, LI BIN, SONG SHIYING, PENG QI
• Format: Patent
• Language: Chinese; English
• Published: 12.04.2022
• 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; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention relates to the technical field of animal virology and animal infectious diseases, and discloses a PCR (Polymerase Chain Reaction) primer for amplifying an N-terminal hypervariable region of an S gene of a porcine epidemic diarrhea virus, a pair of primers is designed in a conserved region obtained by analyzing sequences at two ends according to the N-terminal hypervariable region (V2) of the S gene of the porcine epidemic diarrhea virus, and the pair of primers is used for fragment amplification of V2. And the obtained product can be subjected to subsequent strain typing sequencing. The method further provides the whole process of treatment of clinical porcine diarrhea excrement and intestinal tissue samples, extraction of total RNA, reverse transcription, PCR, electrophoresis gel leakage and result judgment. By using the method, 1823bp fragments can be seen in a positive sample. The amplification primer provided by the invention is good in conservative property, and the PCR method is good in sp