Novel gene quantitative detection method composition, kit and method thereof

A detection method and composition technology, applied in biochemical equipment and methods, microbial measurement/testing, recombinant DNA technology, etc., can solve problems such as expensive instruments and reagents, difficulty in meeting the detection needs of on-site or grassroots laboratories, etc.

Inactive Publication Date: 2018-03-16
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this type of technology is sensitive and accurate, it needs to rely on expensive instruments and reagents, and it is difficult to meet the detection needs of on-site or grassroots laboratories

Method used

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  • Novel gene quantitative detection method composition, kit and method thereof
  • Novel gene quantitative detection method composition, kit and method thereof
  • Novel gene quantitative detection method composition, kit and method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033]This example verifies the specificity, sensitivity and linear correlation of pig and bovine specific padlock probes.

[0034] The probe sequence was synthesized by IDT Biological Co., Ltd. of the United States. The 5' end of the padlock lock probe was connected to a phosphate, the 5' end of the capture probe was labeled with biotin, and the 5' end of the detection probe was labeled with HRPMT. See the table below for the specific sequence.

[0035] name

sequence

illustrate

GHpadlock1cow probe

5' AGCTTTGGGCTTTAGATCCTCAATGCTGCTGCTG TACTACCAATAGAGGTACAGTGAATGCGAGTCCGTCTCGATGGATGTGTTCAG 3' (SEQ ID No. 1)

Phosphate group attached to the 5' end

GHpadlock1pig probe

5' AACCTTGGGCTTTGCTTCTAATCCTCAATGCTGCTGCTTTACTAGTGCTGGATGATCGTCCCTTCTCGGATGGGGCACC 3' (SEQ ID No. 2)

Phosphate group attached to the 5' end

Cytbpadlock1cow probe

5' ATCCGATACATACACGCCAAGAAATCCTCAATGCTGCTAGGTTCCAGTGAATGCGAGTCCGTCTCAGAAAGAACTACGGCTGAATC 3' (SEQ I...

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Abstract

The invention relates to a novel object gene quantitative detection method padlock-RCA-ELISA method. The invention also relates to an oligonucleotide probe composition for relative quantitative detection on different animal source growth hormone genes in animal tissues (pig and cattle) and relative quantitative detection (pig and cattle) on mitochondrial cytochrome b gene by using the method. Theinvention also relates to a padlock-RCA-ELISA kit containing the composition. By using the method, quantitative detection can be carried out on a target gene in a simple, rapid, and accurate mode.

Description

technical field [0001] The invention belongs to the field of biotechnology. Specifically, the invention relates to a novel gene quantitative detection method-padlock-RCA-ELISA method, and using the method to perform relative quantification of growth hormone genes from different animal sources in animal tissues (pig and bovine ) and oligonucleotide probe compositions for relative quantification (porcine and bovine) of mitochondrial cytochrome genes. It also relates to a padlock-RCA-ELISA kit comprising said composition. Background technique [0002] Gene quantitative detection is widely used in medical diagnosis, infectious pathogen monitoring, forensic medicine, and food safety. The current common method is real-time fluorescent quantitative PCR, and the latest technology room digital PCR. These methods all need to be based on PCR technology, and the signal detection is realized by amplifying the target gene in the molecular copy number. Although this type of technology i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q2531/125
Inventor 吴亚君陈磊陈颖刘鸣畅杨艳歌
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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