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Free DNA sequence derived from echinococcus granulosus and application of free DNA sequence

A technology of Echinococcus granulosus and DNA sequence, which is applied in the fields of biotechnology and medicine and achieves the effects of good application prospect, high specificity and sensitivity, and high efficiency

Active Publication Date: 2020-10-02
中国疾病预防控制中心寄生虫病预防控制所国家热带病研究中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] There are no publications on the use of cell-free DNA sequences of Echinococcus granulosus for the detection and diagnosis of echinococcosis (or echinococcosis)

Method used

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  • Free DNA sequence derived from echinococcus granulosus and application of free DNA sequence
  • Free DNA sequence derived from echinococcus granulosus and application of free DNA sequence
  • Free DNA sequence derived from echinococcus granulosus and application of free DNA sequence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Acquisition of Free DNA Sequences from Echinococcus granulosus

[0042] 1.1 Source of resequencing plasma samples

[0043] Fresh plasma was collected from 3 patients with cystic echinococcosis (from Qinghai Provincial People’s Hospital). The patients with echinococcosis were diagnosed by clinical imaging and all met the indications for surgery. Blood samples were collected before the operation. A patient with cystic echinococcosis.

[0044] 1.2 Resequencing

[0045] 1.2.1 Cell-free DNA extraction from plasma samples

[0046] The free DNA from the fresh plasma of the echinococcosis patients was extracted by QIAamp Circulating Nucleic Acid Kit from QIAGEN Company. After the electrophoresis test is qualified, the library will be built.

[0047] 1.2.2 Library construction

[0048] Qualified DNA samples were randomly cut into 350bp fragments with a Covaris ultrasonic nucleic acid shearing instrument, and the TruSeq DNA LT Sample Prep Kit was used to build a li...

Embodiment 2

[0058] Example 2 Verification of Cell-Free DNA Sequence Sensitivity and Specificity

[0059] 2.1 Standard blood samples as positive and negative controls

[0060] 20 plasma samples from patients with cystic echinococcosis (taken from Qinghai Provincial People’s Hospital). The patients with hydatid disease were diagnosed by clinical imaging and all met the indications for surgery. Blood samples were collected before the operation. For worm patients, the 20 plasma samples were extracted from the free DNA using the QIAamp Circulating Nucleic Acid Kit from QIAGEN as a positive control;

[0061] 20 healthy people's plasma (taken from Qinghai Provincial People's Hospital) came from the same echinococcosis endemic area as the echinococcosis patients. There were no abnormal indicators after physical examination. The blood samples were tested by parasite ELISA and were all negative. The 20 plasma samples were used Free DNA was extracted with the QIAamp Circulating Nucleic Acid Kit kit...

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Abstract

The invention discloses a free DNA sequence derived from echinococcus granulosus. The length of the free DNA sequence is equal to 107 basic groups and is represented by SEQ ID NO.24. Besides, the invention further discloses a screening method of the free DNA sequence derived from the echinococcus granulosus. The screening method comprises the following steps: (S1) acquiring a large number of freeDNA sequences from a blood sample of a patient with cytic echinococcosis by virtue of a re-sequencing technique; (S2) carrying out comparative analysis by virtue of a bioinformatics method so as to screen out echinococcus granulosus-sourced free DNA sequences; and (S3) carrying out verification and screening in patients with the echinococcosis and healthy people through a fluorescence quantitativePCR method, so as to obtain the free DNA sequence derived from the echinococcus granulosus. Besides, the invention further discloses application of the free DNA sequence derived from the echinococcusgranulosus in preparation of products for diagnosing and detecting the echinococcosis. A detection result shows that the free DNA sequence has relatively high sensitivity and specificity in detectionof the patient with the echinococcosis and has excellent application prospects in the diagnosis and detection fields of the echinococcosis.

Description

technical field [0001] The invention belongs to the fields of biotechnology and medicine, and is specifically a free DNA sequence derived from Echinococcus granulosus; in addition, the invention also relates to the use of the free DNA sequence in the preparation of echinococcosis (or echinococcosis) detection and diagnosis products in the application. This invention is funded by National Natural Science Foundation of China (81501771, 81601792, 81772224). Background technique [0002] The larvae of Echinococcus spp. can cause echinococcosis, commonly known as hydatid disease, when parasitic on human body. Insect disease is a parasitic disease that my country focuses on. Among them, the echinococcosis caused by Echinococcus granulosus is cystic echinococcosis, accounting for 98% of the total number of echinococcosis. my country is the country with the most severe echinococcosis epidemic. There are 370 echinococcosis endemic counties in the country, and the endemic area accou...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6806C12N15/11
CPCC12Q1/6888C12Q1/6806C12Q2531/113C12Q2563/107Y02A50/30
Inventor 王莹伍卫平韩秀敏张璟曹建平沈玉娟
Owner 中国疾病预防控制中心寄生虫病预防控制所国家热带病研究中心
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