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SNORD57 detection kit for screening and diagnosing intestinal polyp, intestinal adenoma and/or intestinal cancer

A detection kit and sequence technology, applied in the direction of microbial determination/inspection, DNA/RNA fragmentation, recombinant DNA technology, etc. Suitable for a wide range of samples, good specificity, and high amplification efficiency

Active Publication Date: 2022-05-06
广州瑞熹生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There are few domestic and foreign reports on the detection of SNORD57, but the direct design of corresponding primers and probes for SNORD57 will cause non-specific amplification
Currently there is no fast, simple and effective detection method for SNORD57

Method used

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  • SNORD57 detection kit for screening and diagnosing intestinal polyp, intestinal adenoma and/or intestinal cancer
  • SNORD57 detection kit for screening and diagnosing intestinal polyp, intestinal adenoma and/or intestinal cancer
  • SNORD57 detection kit for screening and diagnosing intestinal polyp, intestinal adenoma and/or intestinal cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Primer and probe design

[0045] According to GenBank, the nucleotide sequence of SNORD57 was obtained, as shown in SEQ ID NO.1, and its reverse complementary sequence was shown in SEQ ID NO.2. The SNORD57 refers to nucleolar RNA, and the name is SNORD57 or C / D box 57. The BLAST function analysis of NCBI found that the SNORD57 sequence has a certain homology with the human genome, indicating that the corresponding primers and probes cannot be directly designed for SNORD57, which will cause non-specific amplification. To avoid this situation, unique reverse transcription primers were designed, the reverse transcription product of which contained the SNORD57 sequence without causing non-specificity in subsequent PCR. figure 1 Schematic diagram of the principle of primer design.

[0046] 1. Reverse transcription primer

[0047] The 3' end of the SNORD57 reverse transcription primer has 9 bases complementary to the 3' end sequence of the SNORD57 gene, which can...

Embodiment 2

[0058] Example 2 A SNORD57 Detection Kit for Screening and Diagnosing Intestinal Polyps, Intestinal Adenomas and / or Intestinal Cancers

[0059] 1. Composition

[0060] 1 tube of reverse transcription reaction solution: including reverse transcription buffer and reverse transcription primer whose nucleotide sequence designed in Example 1 is shown in SEQ ID NO. 3, and the final concentration of the reaction system of reverse transcription primer is 0.1 μmol / L.

[0061] 1 tube of reverse transcription reaction enzyme mixture: contains reverse transcriptase, RNase inhibitor and dNTPs.

[0062] 1 tube of PCR reaction solution: contains Taq enzyme, UNG enzyme, dNTPs (including dUTP) and PCR buffer;

[0063] It also includes the PCR primer set designed in Example 1, that is, the forward primer whose nucleotide sequence is shown in SEQ ID NO.4, the reverse primer whose nucleotide sequence is shown in SEQ ID NO.5, and a nucleoside The specific probe whose acid sequence is shown in SE...

Embodiment 3

[0083] Example 3 Detection of the amplification efficiency of the kit

[0084] 1. Experimental method

[0085] Select the determined SNORD57 RNA and dilute it into 4 gradients with 10-fold nuclease-free water to prepare a quantitative standard (concentrations are 1 × 10 6 copies / mL, 1×10 5 copies / mL, 1×10 4 copies / mL, 1×10 3 copies / mL), use the test kit of embodiment 2 to detect, simultaneously use negative, positive quality control substance to carry out quality control.

[0086] 2. Experimental results

[0087] There is no S-shaped amplification curve in the FAM of the negative quality control product; the amplification curves of the FAM channel of the positive quality control product are all obvious S-shaped curves, and the Ct value of the positive quality control product is 28.56. Both negative and positive quality control materials meet the quality control requirements of the kit, so the test results of the samples to be tested are valid.

[0088] The amplification ...

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Abstract

According to the SNORD57 detection kit for screening and diagnosing the intestinal polyp, the intestinal adenoma and / or the intestinal cancer, the detection kit comprises an SNORD57 reverse transcription primer for detecting the SNORD57 and / or a primer probe composition for detecting the SNORD57, and the primer probe composition for detecting the SNORD57 comprises an SNORD57 forward primer, an SNORD57 reverse primer and an SNORD57 probe. The specific reverse transcription primer and the primer group are provided for the finger kernel RNA SNORD57, and the specific reverse transcription primer and the primer group have the advantages of high amplification efficiency, good specificity and the like. According to the invention, semi-automatic detection of SNORD57 is realized, the speed is high, the sensitivity is high, and the applicable sample range is wide.

Description

technical field [0001] The invention relates to the technical field of molecular detection, in particular to a SNORD57 detection kit for screening and diagnosis of intestinal polyps, intestinal adenomas and / or intestinal cancer. Background technique [0002] Nucleolar RNA (snoRNA) is a kind of small non-coding RNA widely present in the nucleolus of eukaryotic cells. Molecules that perform post-transcriptional modifications. The expression of snoRNA may be regulated by host genes, CNVs and DNA methylation. Substantial evidence indicates the importance of snoRNAs in the development of cancer, for example, SNORD78 is overexpressed in non-small cell lung cancer and prostate cancer, and SNORDSOA / B, which can directly bind and inhibit K-Ras, is deleted in multiple cancer types ; SNORA42 expression is an independent prognostic factor for increased overall survival time in cancer patients. SNORA55 silencing in prostate cancer cell lines significantly inhibited cell proliferation ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6883C12Q1/686C12N15/11
CPCC12Q1/6886C12Q1/6883C12Q1/686C12Q2561/113C12Q2563/107C12Q2521/107C12Q2527/127C12Q2521/531
Inventor 周其伟马强李明
Owner 广州瑞熹生物科技有限公司
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