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Rapid detection method and kit for indicating total glycoside content range of cistanche deserticola raw drug

A detection kit and detection method technology, which is applied in the field of biotechnology detection, can solve problems such as complexity, time-consuming, cumbersome experimental procedures, etc., and achieve the effects of strong specificity, high sensitivity, and shortened detection cycle

Inactive Publication Date: 2016-01-27
BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for the detection of the phenylethanol glycoside content of Cistanche deserticola, the above-mentioned main detection methods all require the excavated Cistanche deserticola to be dried for at least half a month, and then ground after drying. This process takes a long time and is The experimental process is cumbersome and complicated

Method used

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  • Rapid detection method and kit for indicating total glycoside content range of cistanche deserticola raw drug
  • Rapid detection method and kit for indicating total glycoside content range of cistanche deserticola raw drug
  • Rapid detection method and kit for indicating total glycoside content range of cistanche deserticola raw drug

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Experimental program
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Effect test

Embodiment 1

[0085] Embodiment 1: Test the total glycoside content of Cistanche deserticola in random sampling (sample collection place: Inner Mongolia Left Banner).

[0086] (1) RNA extraction;

[0087] (2) RNA purification;

[0088] (3) reverse transcription reaction;

[0089] (4) RT-qPCR reaction;

[0090](5) Calculate the total glycoside content range according to the PCR results: According to the experiment, the CT values ​​of the four genes are respectively: SEQ1 is 33.90369415, SEQ2 is 27.05919711, SEQ3 is 34.07738368, and SEQ4 is 30.52809588, and the calculated n 1 , n 2 , n 3 :

[0091] no 1 = 2 -(CT(SEQ1)-CT(SEQ4)) =0.096348214

[0092] no 2 = 2 -(CT(SEQ2)-CT(SEQ4)) =11.0724208

[0093] no 2 = 2 -(CT(SEQ3)-CT(SEQ4)) =0.085419674

[0094] Calculated according to formula (Ⅱ) to get P 1 ,P 2 ,P 2 , using the data in Table 5, in the left flag: N 11 =0.02,N 12 =1.63,N 13 =0.01,N 21 =0.01,N 22 =4.50,N 23 =0.04,N 21 = 0, N 32 =9.64,N 33 =0.01, compare 3 P at th...

Embodiment 2

[0100] Example 2: Test the total glycoside content of Cistanche deserticola in random samples (sample collection place: Right Banner, Inner Mongolia).

[0101] (1) RNA extraction;

[0102] (2) RNA purification;

[0103] (3) reverse transcription reaction;

[0104] (4) RT-qPCR reaction;

[0105] (5) Calculate the range of total glycoside content according to the PCR results: According to the experiment, the CT values ​​of the four genes are respectively: SEQ1 is 32.59574763, SEQ2 is 28.58986219, SEQ3 is 34.18855667, and SEQ4 is 29.99904887, and the calculated n 1 , n 2 , n 3 :

[0106] no 1 = 2 -(CT(SEQ1)-CT(SEQ4)) =0.096348214

[0107] no 2 = 2 -(CT(SEQ2)-CT(SEQ4)) =11.0724208

[0108] no 2 = 2 -(CT(SEQ3)-CT(SEQ4)) =0.085419674

[0109] Calculated according to formula (Ⅱ) to get P 1 ,P 2 ,P 2 , using the data in Table 4, in the right flag: N 11 =0.04,N 12 =3.33,N 13 =0.38,N 21 =0.01,N 22 =8.02,N 23 =0.06,N 21 = 0, N 32 =21.33,N 33 =0.02, compare 3 P a...

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Abstract

The invention discloses a rapid detection method and kit for indicating the total glycoside content range of cistanche deserticola raw drug. According to the invention, real-time fluorescence quantitative PCR method is employed to determine the expression values of cistanche deserticola raw drug's four genes, and the employed primer sequences are as the following: SEQ1; SEQ2; SEQ3; and SEQ4; wherein the gene SEQ2 is positively correlated with the total glycoside content, the gene SEQ1 is negatively correlated with the gene SEQ3; and the gene SEQ4 is a reference gene. The method provided by the invention utilizes the next generation sequencing technology and HPLC technology to identify the three genes varying cooperatively with the total glycoside content of cistanche deserticola, and utilizes real-time fluorescence quantitative PCR to rapidly indicate the content of main medicinal components of the raw drug, thus providing guidance and standard reference for cistanche deserticola raw drug grading, efficient product development and other applications, and has the characteristics of strong specificity, high sensitivity, and high efficiency and speed.

Description

technical field [0001] The invention belongs to the field of biotechnology detection, and relates to a rapid detection method and a kit for indicating the content range of total glycosides in raw medicinal materials of Cistanche deserticola. Background technique [0002] Cistanche deserticola, known as "desert ginseng", has extremely high medicinal value and is widely used in traditional Chinese medicinal materials. It was recorded in "Shen Nong's Herbal Classic" more than 1,800 years ago. Cistanche is sweet in taste and warm in nature. It has the functions of nourishing kidney and strengthening yang, enhancing immunity, nourishing blood and moistening dryness, prolonging life, protecting liver and protecting liver. Cistanche Herba Cistanche is used for both medicine and food, and it has been regarded as a treasure by countries in the Western Regions as a tribute to the imperial court in history. Cistanche's suitable food groups mainly include: those with physical deficienc...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2561/113C12Q2561/101C12Q2545/114C12Q2531/113
Inventor 宋述慧陈婷婷李语丽王西亮姚富文
Owner BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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