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Detection primer set for LINE-1 and application of detection primer group

A LINE-1, primer set technology, applied in the field of genes, can solve the problems of high cost, poor adaptability, lack of specificity and sensitivity, etc., and achieve the effect of good specificity

Pending Publication Date: 2020-08-25
邹畅
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Problems solved by technology

However, there are some limitations in imaging detection: it is difficult to detect tumors at an early stage, and some imaging detections have disadvantages such as poor adaptability (such as colonoscopy) and high cost
However, although there are many copies of LINE-1 in the human genome, more than 95% of the 5' end of LINE-1 is deleted, and 10% of LINE-1 is also rearranged, so it has full-length sequence and transposition activity LINE-1 is very rare, and which fragments of LINE-1 can be used as detection targets for tumor screening requires extensive research on LINE-1 related research and verification with a large number of samples
Moreover, the detection effect of a single primer may not have sufficient specificity and sensitivity, and the combination of two or even more pairs of primers may have a better effect. How to screen out a suitable primer combination is a difficulty.

Method used

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  • Detection primer set for LINE-1 and application of detection primer group
  • Detection primer set for LINE-1 and application of detection primer group
  • Detection primer set for LINE-1 and application of detection primer group

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Embodiment Construction

[0062] The technical solution of the present invention will be further described below in combination with experiments.

[0063] Specificity comparison:

[0064] Based on the published LINE1 sequence, 2 pairs of long-segment amplification primers and 2 pairs of short-segment amplification primers were designed, and two pairs of primers (one long and one short) reported in previous literatures (Table 1) were selected for comparison. It was found that LINE1- The two pairs of primers S-A and LINE1-L-A have the best amplification efficiency and specificity, so LINE1-S-A and LINE1-L-A are used in subsequent detection experiments.

[0065] Table 1: Design primers and primer sequences in the literature

[0066]

[0067] Note: The document marked with "*" is: Song Xianrang. The method for measuring the content and integrity of ctDNA with LINE1 gene as the detection object. The SEQ ID NO. numbers of the primer sequences are 1-12 from top to bottom.

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Abstract

The invention discloses a detection primer set for LINE-1 and application of the detection primer set. The primer set comprises primers as shown in SEQ ID NO. 1, 2, 7 and 8. The primer set provided byembodiments of the invention has good specificity and excellent amplification efficiency. The primer set can be used for early screening, recurrence diagnosis and treatment monitoring of various tumors, and has the advantages of simplicity, convenience, rapidness and accuracy.

Description

technical field [0001] The invention relates to the gene field, in particular to a detection primer set for LINE-1 and its application. Background technique [0002] Cancer is a major disease that plagues mankind. According to statistics, there were about 18.1 million new cancer cases worldwide in 2018, and another 970 cancer patients died. In China in 2018, there were about 3.8 million new cancer cases and about 2.29 million deaths, and the overall cancer incidence rate increased by about 3.2%. Cancer has become one of the four major chronic diseases in my country, seriously affecting people's health. Early screening and diagnosis of cancer can effectively reduce cancer mortality. Effective monitoring of treatment effect and recurrence can provide a reliable reference for precise treatment of cancer patients. The current gold standard for cancer diagnosis is imaging testing. However, there are some limitations in imaging detection: it is difficult to detect tumors at an...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/11
CPCC12Q1/6886C12Q1/6851C12Q2600/118C12Q2600/156C12Q2531/113C12Q2525/151C12Q2537/165
Inventor 邹畅陈利鹏赵盼
Owner 邹畅
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