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DNA bar code standard detection fragment for identifying vespa soror and application of detection fragment

A standard detection and barcoding technology, which is applied in the field of DNA barcoding standard detection fragments for identification of the yellow striped hornet, and achieves the effects of improving identification efficiency, high repeatability and fast speed.

Active Publication Date: 2020-09-08
DALI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no effective DNA barcode identification method with high accuracy to identify the yellow-striped hornet.

Method used

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  • DNA bar code standard detection fragment for identifying vespa soror and application of detection fragment
  • DNA bar code standard detection fragment for identifying vespa soror and application of detection fragment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 is used to identify the acquisition of the DNA barcode standard gene sequence of Vespa yellow

[0037] 1. Collection, identification and storage of samples to be tested

[0038] The wasp sample to be tested was purchased in Zengcheng City, Guangdong Province. The sample was identified as Vespa soror by morphological methods by Professor Dong Dazhi of Kunming Institute of Zoology, Chinese Academy of Sciences. After the identification was correct, the samples were soaked in absolute ethanol and stored in a -20°C freezer.

[0039] 2. Preparation of experimental instruments and reagents

[0040] 2.1 Experimental instruments and reagents

[0041] HWS12 constant temperature electric heating constant temperature water bath (Shanghai Yiheng Technology Co., Ltd., serial number 131022035);

[0042] SIGMA 1-14ED centrifuge (Sigma, Germany, serial number 133828);

[0043] DYCP-31DN electrophoresis instrument (Beijing Liuyi Instrument Factory, batch number 20130816); ...

Embodiment 2

[0106] Example 2 Identification of samples of different forms and processed products of Vespa yellow stripes

[0107] 1. Collection and storage of the samples to be tested: take the pupae and adults of Vespa japonica purchased in Dehong City, Yunnan Province as the samples to be tested, and take part of the adult worms and quickly dry them in an oven to become adult processed products. Vespa adults purchased in Zengcheng City, Guangdong Province were used as positive control samples. All samples were identified as Vespa soror by Prof. Dong Dazhi, Kunming Institute of Zoology, Chinese Academy of Sciences according to morphological methods. After the identification was correct, the samples were soaked in absolute ethanol and stored in a -20°C freezer.

[0108] 2. Preparation of experimental instruments and reagents: the same as in Example 1.

[0109] 3. DNA extraction and template preparation: first, pretreat the positive control sample and the sample to be tested. Sample pre...

Embodiment 3

[0115] The specificity detection of the DNA barcode molecular identification method of embodiment 3 Vespa yellow stripes

[0116] 1. Collection and storage of samples to be tested

[0117] The samples to be tested were the medicinal materials of Vespa japonica purchased in Dehong City, Yunnan Province, and the medicinal materials of Periplaneta americana purchased in Baoshan City, Yunnan Province. The samples were identified as Vespa soror and Periplaneta americana Linn. by Prof. Dong Dazhi, Kunming Institute of Zoology, Chinese Academy of Sciences, and Prof. Yang Zizhong, Dali University. The morphologically identified Vespa adults were used as positive control samples. Wipe the sample to be tested with 75% alcohol for disinfection, take a part of the head or chest muscle tissue of the sample to be tested after disinfection, put it in a 1.5mL centrifuge tube, then soak the muscle tissue with absolute ethanol and store it in a -20°C refrigerator.

[0118] 2. Preparation of e...

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Abstract

The invention discloses a DNA bar code standard detection fragment for identifying vespa soror and application of the detection fragment, and belongs to the field of biological species identification.The nucleotide sequence of the DNA bar code standard detection fragment is as shown in SEQ ID No. 1, and the invention also provides a method for molecular identification of insect medicinal materialvespa soror by using the gene sequence and a PCR amplification technology. A PCR product is verified through agarose gel electrophoresis and then subjected to sequencing, a sequencing result is subjected to manual proofreading and sequence splicing and is compared with a COI gene sequence SEQ ID NO.1 disclosed by the invention, and therefore whether a sample to be detected is vespa soror or not is judged. According to the DNA molecular identification technology, morphological identification does not need to be carried out on insect medicinal material scorpions, quick, accurate and reliable identification on fresh or dry medicinal materials, whole worms or part of worms, pupae or larvae or adults is achieved, and the safety of taking the vespa soror as a medicinal material is guaranteed.

Description

technical field [0001] The invention relates to the field of identification of biological species, in particular to a DNA barcode standard detection fragment for identifying Vespa yellow-striped and its application. Background technique [0002] Vespas, commonly known as wasps or wasps, are the general term for insects in the superfamily Vespoidea of ​​the order Hymenoptera, which mainly includes seven families including Vespidae, Polistidae and Eumenidae. Vespa is not only a widely used resource insect, but also has high medicinal value. It is used as a medicinal insect in folk ethnic medicine prescriptions in Southwest, Southeast, Taiwan and other regions of my country. The bee larvae, adults, bee venom and honeycomb can be used as medicine, and the drug names are respectively bumble bee, bumble bee larva, bee venom and dew hive. Vespa is sweet and pungent in taste, warm in nature, and mainly treats rheumatic arthralgia. Its adults and larvae have the effect of expelling ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2600/166C12Q2565/125C12Q2535/122C12Q2545/113C12Q2537/165
Inventor 张成桂许凯歌陈壮志杨永寿杨红童建新赵昱葛建
Owner DALI UNIV
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