Novel bio-barcode detection method for ricin

Abstract

The invention discloses a high-sensitivity novel bio-barcode detection method for ricin, which is used for the detection of ricin protein. According to the novel bio-barcode detection method, a marked bar code DNA strand is improved to be a single strand on the basis of the conventional bio-barcode detection method; the marked bar code DNA strand is prolonged until the marked bar code DNA strand can be detected by a fluorescent quantitative PCR / TaqMan fluorescent probe; and a bar code DNA strand in a sandwich reaction compound does not need to be dissociated and is directly used for performing qualitative detection on the bar code RNA strand in the sandwich reaction compound by a TaqMan fluorescent probe-based real-time fluorescent quantitative PCR method or the conventional PCR method. The method has great practical significance for the high sensitivity detection of the ricin protein and has wide application prospect.

Description

technical field [0001] The invention relates to a detection method and its application in the field of biotechnology, in particular to a novel biological barcode detection method for high-sensitivity detection of ricin. Background technique [0002] Ricin is a plant glycoprotein extracted from the endosperm of castor seeds, which refers to four toxic substances of ricin, ricinine, ricin allergen and hemagglutinin, among which ricin It is the most toxic and is an important biological warfare agent and source of bioterrorism. In pharmaceutical research, ricin, also known as ricin, is a ribosome-inactivating protein. Ricin is an odorless white powder or crystalline solid. It consists of two polypeptide chains A and B covalently linked by disulfide bonds to form a heterodimer. Among them, chain A is the effector chain with a molecular weight of about 30kDa or 32kDa. It has glycosidase Activity, inhibiting protein synthesis by destroying the 60S subunit of the ribosome, resulti...

AI Technical Summary

Problems solved by technology

In the PCR reaction, due to factors such as the aggregation of templates, reagents, and pyrophosphate molecules, the polymerase reaction is affected, and eventually the PCR reaction no longer proceeds exponentially and enters a "plateau phase", and some reactions have more end products than others , so the quantification of the endpoint PCR reaction method is inaccurate; secondly, the endpoint PCR is prone to cross-contamination, resulting in false positives

The DNA marked by traditional BCA technology is double-stranded, with a length of about 48bp. Not only is the operation cumbersome and the detection process is lengthy, but it also indirectly affects the detection sensitivity. In addition, it is not suitable for real-time fluorescent quantitative PCR detection based on TaqMan probes. Sample templates for precise quantitative analysis

The chip detection of barcode DNA is prone to false positives due to the shortcomings of silver staining reagents, and it is not easy to define negative and positive, and the staining results vary greatly due to time and environmental factors

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