Detection method of chIRF3 fluorescent quantitation RT-PCR

• Main Authors: HUANG HAIPEI, LI SHANGQUAN, SHEN WEIWEI, CHEN BOWEN, CHEN YANYAN, HE XIUMIAO, LIU YUJING
• Format: Patent
• Language: Chinese; English
• Published: 17.09.2019
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; VINEGAR; WINE

The invention discloses a detection method of chIRF3 fluorescent quantitation RT-PCR. The detection method comprises the following steps of performing primer design, performing total RNA extraction, performing chIRF3 gene amplification, enabling an amplification product to be connected with a pUC-T carrier to construct a plasmid standard product, performing experiment on a fluorescent quantitation RT-PCR amplification system and condition, sensitivity, repeatability and specificity, and performing chIRF3 detection on Sanhuang chickens infected with infectious bursal disease viruses. According to the experiment, a primer is designed according to a chIRF3 gene sequence, firstly a target fragment is amplified by an ordinary PCR method, the size of a chIRF3 fragment is 146bp, fluorescent quantitation PCR detection is performed by using a recombinant of the amplified fragment and amplified plasmid as a standard product, and the properties of the method are evaluated. The correlation coefficient of the established