Swine-derived reference gene and application thereof in internal control nucleic acid test strip

Abstract

The invention discloses a reference gene primer and a probe for detecting a swine-derived sample. The reference gene primer and probe respectively have nucleic acid sequences shown in SEQ ID NO:1-3. The reference gene primer can meet requirement of internal control nucleic acid detection when a swine-derived sample is detected, and avoid false negative results. The invention also discloses an application of the primer in an internal control nucleic acid test strip, and specific steps are as follows: designing a primer and a probe for pathogenic conservative genes, taking extracted sample DNA as a template, carrying out PCR amplification on the pathogenic detection primer and the reference primer at the same time, adding the reference probe and the pathogenic probe after the reaction is finished, detecting the PCR product of the sample to be detected by using the internal control nucleic acid test strip, and reading detection results. The invention can better meet requirements of fielddetection, and can be used for on-site detection of import and export quarantine, primary veterinary laboratories, livestock farms and the like.

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to a pig-derived internal reference gene and the application of the internal control nucleic acid test strip. Background technique [0002] Molecular biology detection methods are widely used in the detection of diseases due to their fast detection, high sensitivity, and good specificity. False positives and false negatives are potential problems that plague molecular biology testing, and false negatives are more serious than false positives in clinical testing. This is because the cause of false positives is mostly the contamination of amplification products, which can be solved by adding UNG enzyme and nucleic acid removal reagents, and sub-regional detection. However, false negatives are different. It involves the entire detection process, whether the experimental equipment is calibrated, whether the reagents are good or bad, and the differences between batches, the skill of the o...

AI Technical Summary

Problems solved by technology

However, false negatives are different. It involves the entire detection process, whether the experimental equipment is calibrated, whether the reagents are good or bad, and the differences between batches, the skill of the operator, and the amplification inhibitors in the sample will all cause false negative results.

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