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Primer probe combination and detection method for detecting gambogia gambosa and agaricus bisporus

A technology of Apricot bisporus and Apricot bisporus, which is applied in the field of primer probe combination and detection for detection of Apricot bisporus and Apricot bisporus, and can solve problems such as authenticity identification of Apricot bisporus and Apricot bisporus, Achieve the effects of saving testing procedures and time, improving testing accuracy, and reducing testing costs

Active Publication Date: 2022-08-05
XILINGOL VOCATIONAL COLLEGE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to solve the problem existing in the prior art that it is difficult to identify the authenticity of Apricot apricot and Agaricus bisporus, and provide a combination of primers and probes and a detection method for detecting Apricot apricot and Agaricus bisporus , can detect Apricot apricot and Agaricus bisporus in the same tube, and perform accurate qualitative and quantitative detection on both

Method used

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  • Primer probe combination and detection method for detecting gambogia gambosa and agaricus bisporus
  • Primer probe combination and detection method for detecting gambogia gambosa and agaricus bisporus
  • Primer probe combination and detection method for detecting gambogia gambosa and agaricus bisporus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Sequence design of primers and probes

[0024] Comparison of Apricot mushrooms, Agaricus bisporus, Mongolian white mushrooms, Xiuzhen mushrooms, Shiitake mushrooms, Pleurotus eryngii, Enoki mushrooms, Seafood mushrooms, White jade mushrooms, Big white pile mushrooms, Money mushrooms, Cordyceps mushrooms, Beef liver mushrooms, Matsutake mushrooms, sheep The ITS genomes of 17 edible fungi such as Trichosanthes, Chashu and Agaricus, and the ITS genome sequences of 10 varieties or strains were selected for each edible fungi. The above-mentioned 170 sequences were compared by bioinformatics software, and the conserved and specific sequences of Apricot mushroom and Agaricus bisporus were screened out, and primer design software was used to design primers and probes. The sequences of the forward and reverse primers are shown in SEQ ID No.1 and SEQ ID No.2; the sequences of the Apricot mushroom probe, Agaricus bisporus probe and quality control probe are shown in sequ...

Embodiment 2

[0026] Example 2 Establishment of qualitative and quantitative detection methods for Apricot moniliformes and Agaricus bisporus

[0027] (1) Extract the DNA of the mushroom sample to be tested.

[0028] (2) Detect the concentration and quality of sample DNA, and dilute the concentration to 100-200ng / μL.

[0029] (3) Utilize primers and probes prepared in Example 1 to carry out amplification detection on the diluted DNA, utilize Apricot moniliformes and Agaricus bisporus positive standards to do positive controls, utilize sterilized deionized water to do negative controls, utilize DNA The extracted blank control was used as the control group of the extraction method, the Real-time PCR reaction system was shown in Table 1, and the Real-time PCR amplification parameters were shown in Table 2.

[0030] Table 1Real-time PCR reaction system (simultaneous detection of apricot mushroom, Agaricus bisporus and quality control)

[0031]

[0032]

[0033] Table 2 Real-time PCR amp...

Embodiment 3

[0040] Embodiment 3 carries out the specific identification test of Apricot mushroom and Agaricus bisporus to 17 kinds of mushrooms

[0041] Using the primers and probes in Example 1, according to the detection method of Example 2, 17 kinds of mushrooms (Agaricus bisporus, Apricot mushroom, Mongolian white mushroom, Xiuzhen mushroom, Shiitake mushroom, Pleurotus eryngii, Flammulina velutipes, Seafood mushroom, White jade mushroom, large white pile mushroom, golden mushroom, cordyceps mushroom, porcini mushroom, pine mushroom, morel mushroom, tea tree mushroom and Agaricus) were detected by qPCR. The test results are attached image 3 , attached Figure 4 and shown in Table 4. attached image 3 Amplification curve appeared in Apricot serrata, but no amplification curve appeared in other mushrooms, indicating that the primers and probes of Apricot serrata were specific. attached Figure 4 Amplification curve appeared in Agaricus bisporus, and no amplification curve appeared...

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Abstract

The invention discloses a primer probe combination for detecting gambosa gambosa and agaricus bisporus. The primer probe combination comprises forward and reverse primers, a gambosa gambosa probe, an agaricus bisporus probe and a quality control probe, the sequences of the forward primer and the reverse primer are as shown in SEQ ID No. 1 and SEQ ID No. 2; the sequences of the gambosa gambosa probe, the agaricus bisporus probe and the quality control probe are shown as SEQ ID No. 3 to SEQ ID No. 5 in sequence. The invention also discloses a method for detecting the gambosa gambosa and the agaricus bisporus by using the primer probe combination. The primer and probe combination is high in specificity and sensitivity, qualitative and quantitative detection of the gambosum gambosum and the agaricus bisporus is achieved, and the detection accuracy of the gambosum gambosum and the agaricus bisporus is remarkably improved; the method can also be used for performing same-tube detection on the gambosa gambosa, the agaricus bisporus and quality control, so that the detection process and time are effectively saved, and the detection cost is remarkably reduced.

Description

technical field [0001] The present invention relates to the technical field of food detection, in particular to a primer-probe combination and a detection method for detecting Apricot moniliformes and Agaricus bisporus. Background technique [0002] Agaricus bisporus is also known as white mushroom, mushroom, mushroom, foreign mushroom, etc. Agaricus bisporus fruiting body is medium and large, cap width is 5-12 cm, initially hemispherical, later flattened, white, smooth, slightly dry to yellow, and the edge is initially involute. The flesh of the fungus is white, thick, slightly reddish after injury, and has a peculiar smell of mushrooms. The gills are pink at first, then turn brown to dark brown, dense, narrow, free, unequal, stipe 4.5-9cm long, 1.5-3.5cm thick, white, smooth, silky, nearly cylindrical, inside Soft or firm, the fungal ring is single-layered, white, membranous, the middle of the stipe, easy to fall off. Agaricus bisporus has a delicious taste and is an ed...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6851C12N15/11C12R1/645
CPCC12Q1/6895C12Q1/6851C12Q2600/16C12R2001/645C12Q2531/113C12Q2563/107C12Q2537/143C12Q2545/114Y02A50/30
Inventor 郭梁徐伟良李春冬李凤忠刘国强罗建兴乌仁图雅木其勒特格希巴雅尔
Owner XILINGOL VOCATIONAL COLLEGE
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