Method for detecting peanut allergen by virtue of locked nucleic acid molecular beacon and application

A peanut allergen and molecular beacon technology, which is applied in biochemical equipment and methods, and microbial measurement/inspection, etc., can solve the problems of ELISA method false negative, long detection time, cumbersome steps of imprinting method, etc., and reach the detection limit Low cost, reduced operation steps, good specificity

Active Publication Date: 2017-12-15
ZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY
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Problems solved by technology

However, the steps are cumbersome, the detection takes a long time, and specific primary and secondary antibodies are required, resulting in high cost
[0007] In summary, the technical problem of the existing technology is that the detected protein may change during the food processing process, resulting in the appearance of false negatives in the ELISA method; and the steps of the blotting method are cumbersome, and the time required for detection is relatively long. Long, and requires specific primary and secondary antibodies, high cost

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  • Method for detecting peanut allergen by virtue of locked nucleic acid molecular beacon and application
  • Method for detecting peanut allergen by virtue of locked nucleic acid molecular beacon and application
  • Method for detecting peanut allergen by virtue of locked nucleic acid molecular beacon and application

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Embodiment Construction

[0026] The present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments, but should not be construed as a limitation of the present invention. The experimental methods in the following examples, unless otherwise specified, are conventional methods, and the materials, reagents, etc. used in the following examples, unless otherwise specified, can be obtained from commercial sources.

[0027] A method for detecting peanut allergens using locked nucleic acid molecular beacons provided by the present invention includes three main steps of DNA extraction, digital PCR and result analysis, specifically including:

[0028] S1, DNA extraction of the sample to be tested

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Abstract

The invention belongs to the technical field of molecular biology, and discloses a method for detecting a peanut allergen by virtue of a locked nucleic acid molecular beacon and application. The method comprises the three main steps of DNA (deoxyribonucleic acid) extraction, digital PCR (polymerase chain reaction) and result analysis, wherein a sequence of a used upstream primer is shown as SEQ ID NO.1, a sequence of a downstream primer is shown as SEQ ID NO.2, and a sequence of a probe is shown as SEQ ID NO.3. Compared with another existing common peanut allergen detection method, the detection method has the advantages that the detection sensitivity and specificity can be prevented from being changed after peanuts are processed into cooked food, the characteristic of low detection limit is achieved, corresponding positive results can be obtained for 1ng / mu g (or 1ng / mu l) in DNA samples, and the accuracy of a detection result reaches 100 percent.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a method and application for detecting peanut allergens by using locked nucleic acid molecular beacons. Background technique [0002] As a new type of nucleic acid derivative, locked nucleic acid (LNA) has the advantages of high affinity, good thermal stability, strong resistance to enzymatic digestion, and non-toxicity in vivo, and can be used as a solid primer like ordinary DNA primers. Therefore, it is widely used in many fields such as antisense therapy and single nucleotide polymorphism (single nucleotide polymorphism, SNP) detection. [0003] Digital PCR is a breakthrough nucleic acid quantitative analysis technology, which has attracted attention and developed rapidly in recent years. This technology is to dilute the nucleic acid template first, divide a sample into tens to tens of thousands of parts, and then distribute them to a large number of ind...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q1/6895C12Q2563/159C12Q2563/107C12Q2545/114C12Q2525/10
Inventor耿尧赵黎明郜连飞
OwnerZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY