Duplex PCR authentication method of cordyceps sinensis original powder

A technology of Cordyceps sinensis and Cordyceps sinensis, which is applied in the field of identification of medicinal materials, can solve the problem of no significant effect in identification of the powder of Cordyceps sinensis, and achieve the effect of low cost, high accuracy, and strong primer specificity

Active Publication Date: 2013-08-07
CHINA NAT RES INST OF FOOD & FERMENTATION IND CO LTD +1
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Problems solved by technology

The above identification methods are mainly used to distinguish Cordyceps sinensis and counterfeit products with similar shapes, but have no significant effect on the authenticity of Cordyceps sinensis grass powder, and it is difficult to play a role in ensuring the quality of Cordyceps sinensis grass powder

Method used

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  • Duplex PCR authentication method of cordyceps sinensis original powder
  • Duplex PCR authentication method of cordyceps sinensis original powder
  • Duplex PCR authentication method of cordyceps sinensis original powder

Examples

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Embodiment 1

[0027] Example 1: Design of PCR primers and detection of samples thereof

[0028] 1 Design of PCR primers

[0029] 1.1 Design of Cordyceps sinensis strain MAT1-2-1 gene-specific primers

[0030] Primer design was carried out according to the Cordyceps sinensis MAT1-2-1 gene sequence information publicly registered on GenBank (see figure 1 ).

[0031] The GenBank accession number of the reference sequence is: FJ654171 (Ophiocordyceps sinensis, MAT1-2-1 gene, complete cds, http: / / www.ncbi.nlm.nih.gov / nuccore / FJ654171).

[0032] An upstream primer was designed at 18-34 and named as CICC-Oph-F1. The sequence is: 5'-GCCAATCCCATCAACAT-3'.

[0033] A downstream primer was designed at 320-337 and named as CICC-Oph-R1. The sequence is: 5'-CACCACCAACGACAAAA-3'.

[0034] 1.2 Design of specific primers for COI gene of Cordyceps sinensis host manta moth

[0035] Primer design was carried out according to the COI gene sequence information of the Cordyceps sinensis host bat moth COI ...

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Abstract

The invention discloses a duplex PCR authentication method of cordyceps sinensis original powder. The method adopts a technical scheme that cordyceps sinensis original powder genome DNA is adopted as a template; with PCR primers designed by the invention, a single-tube duplex PCR reaction is carried out, and characteristic housekeeping genes of a cordyceps sinensis strain and a host hepialus are simultaneously amplified, wherein actual amplified fragments are respectively 320bp and 136bp; and through agarose gel electrophoresis detection, product authenticity is determined. With the method provided by the invention, cordyceps sinensis original powder can be specifically identified, and sample authenticity detection can be completed simply with three steps of DNA extraction, PCR amplification, and electrophoresis detection. The operation is simple and is easy to command, and accuracy is high.

Description

technical field [0001] The invention belongs to the field of identifying the authenticity of medicinal materials, and in particular relates to an authenticating method for the grass powder of Cordyceps sinensis. Background technique [0002] Traditionally, Cordyceps sinensis is mostly decocted or stewed. Since the cell wall of Cordyceps is not broken, the active ingredients are difficult to dissolve, which is not conducive to human body absorption. In order to improve the utilization efficiency of Cordyceps sinensis and increase the absorption of active ingredients of Cordyceps sinensis by the human body, an effective method is to grind Cordyceps sinensis into tablets, capsules, pills, wine and other dosage forms before taking it. In the past, high-quality raw grass was easily accepted by consumers through morphological identification, but after crushing and processing, consumers' acceptance was greatly reduced. The quality control of deep-processed products of Cordyceps si...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12Q1/68
Inventor 程池扎西才吉李辉姚粟刘洋白飞荣
Owner CHINA NAT RES INST OF FOOD & FERMENTATION IND CO LTD
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