Probe identifying three aedes simultaneously on the basis of gene chip and kit

A gene chip and kit technology, applied in the field of gene chip detection and identification probes and kits, can solve the problems of complicated operation, low detection efficiency, low degree of automation, etc., and achieve the effect of strong specificity and high sensitivity

Inactive Publication Date: 2015-11-11
珠海国际旅行卫生保健中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the gene chip technology immobilizes a large number of probes on the support at the same time, it can detect and analyze a large number of sequences in the sample at one time, thereby solving the problem of complicated operation, low degree of automation, small number of operation sequences and detection efficiency of traditional nucleic acid imprinting hybridization technology. Insufficient
At present, there are very few reported technologies for identifying mosquitoes based on gene chips, and only the preliminary research on DNA barcode chips established by Zhao Ming and others can be found. However, the design and development of the chips are mainly through virtual electronic hybridization and verification, and the hybridization between the results and the real objects is still limited. big difference

Method used

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  • Probe identifying three aedes simultaneously on the basis of gene chip and kit
  • Probe identifying three aedes simultaneously on the basis of gene chip and kit
  • Probe identifying three aedes simultaneously on the basis of gene chip and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] [Preparation of Example 1 Gene Chip]

[0023] The preparation method of the present embodiment is as follows:

[0024] 1. Probe design: first download the corresponding gene sequence from the NCBI gene database, and use software to compare the sequences. According to the comparison results, specific oligonucleotide detection probes (Table 1), quality control probes (Table 2) and specific primers (Table 3) were designed at the conserved positions of the gene sequences.

[0025] 2. Synthesis of probes: Add 6 bases T to the 3′ end of each probe and modify the 3′ end amino group as a connecting arm, so that it can be fixed on the chip; except for the amino modification at the 3′ end of the quality control probe , and the 5′ end was simultaneously labeled with biotin.

[0026] 3. Chip preparation: The synthesized probe was diluted to 100uM with deionized water, and 10ul of the probe solution and 10ul of the chip spotting solution were mixed to make the final concentration ...

Embodiment 2

[0045] [Example 2 Detection and identification of Aedes albopictus gene chip]

[0046] The gene chip was prepared according to the method in Example 1, and stored dry at room temperature for future use. Aedes albopictus specimens were taken, and DNA was extracted with an animal tissue genome kit, and the extraction method was operated according to the instructions. PCR amplification and probe hybridization were performed according to the method in Example 1.

[0047] Result analysis: The hybridization results showed that the gene chip prepared in Example 1 had a good specific recognition ability for Aedes albopictus.

Embodiment 3

[0048] [Example 3 Detection and identification of Aedes gene chip in Dongxiang]

[0049]The gene chip was prepared according to the method in Example 1, and stored dry at room temperature for future use. The specimens of Aedes dongxiang were taken, and the DNA was extracted with the animal tissue genome kit, and the extraction method was operated according to the instructions. PCR amplification and probe hybridization were performed according to the method in Example 1.

[0050] Result analysis: The hybridization results showed that the gene chip prepared in Example 1 had a good specific recognition ability for Aedes dongxiang.

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Abstract

The invention belongs to the field of biological detection and identification, and relates to a probe identifying three aedes of an aedes albopictus, an aedes aegypti and an aedes togoi on the basis of a gene chip, and a kit. The invention discloses the probe identifying the three aedes simultaneously on the basis of the gene chip, and the probe comprises the following three probes: the aedes albopictus: CTTTAACACTGCTGCTTTCTAGTTC; the aedes aegypti: GTGCTGAACTTAGCCACCCTGGT; the aedes togoi: AACTCTCCTGCTTTCAAGTAGT. According to gene sequences of the different aedes, conservative positions of the gene sequences are selected and a specific oligonucleotide detection probe is designed; mosquito species identification is carried out by utilizing a biological method; the problems that the number of identification experts is limited, the identification period is long, and the identification efficiency is low and the like in a traditional media mosquito species identification method are overcome; the quick and accurate identification on the three aedes of the aedes albopictus, the aedes aegypti and the aedes togo is realized. According to an experimental detection, the gene chip and the kit provided by the invention are high in specificity, high in sensitivity, and can be well used for media biological identification work of a port.

Description

technical field [0001] The invention belongs to the field of biological detection and identification, and relates to a gene chip detection and identification probe and a kit for Aedes albopictus, Aedes aegypti and Aedes dongxiang. Background technique [0002] Aedes, the collective name for insects of the genus Aedes in the subfamily Culicinae of the family Culicidae. Distributed all over the world, it is the largest genus in the mosquito family, with nearly 1,000 species, and there are more than 100 species in China. Aedes mosquitoes can transmit many diseases, which are harmful to human health. Yellow fever and dengue fever are mainly transmitted by Aedes mosquitoes. Yellow fever is an acute infectious disease caused by yellow fever virus, and Aedes aegypti is the main vector. Dengue fever is an acute infectious disease. It is known that 12 species of Aedes mosquitoes can transmit the disease, but the most important ones are Aedes aegypti and Aedes albopictus. Aedes lar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/16
Inventor 廉国胜汪海波林加燕柯明剑田绿波莫秋华杨泽苏影伍碧梅祝琰
Owner 珠海国际旅行卫生保健中心
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