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Application of reagent for detecting exosome miR-6879-5p in preparation of thyroid cancer metastasis detection kit

A detection kit, detection reagent technology, applied in the field of cancer diagnosis

Active Publication Date: 2022-02-18
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Human miR-6879-5p (MIMAT0027658), a miRNA encoded by human chromosome 11, has few reports about it

Method used

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  • Application of reagent for detecting exosome miR-6879-5p in preparation of thyroid cancer metastasis detection kit
  • Application of reagent for detecting exosome miR-6879-5p in preparation of thyroid cancer metastasis detection kit
  • Application of reagent for detecting exosome miR-6879-5p in preparation of thyroid cancer metastasis detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: test kit of the present invention

[0026] In this embodiment, the PCR detection kit is taken as an example for introduction.

[0027] 1. Composition of the kit

[0028] 1.1 Reverse transcription reagents

[0029] (1) MicroRNA Stem-loop reverse transcription primer

[0030] The sequence (SEQ ID NO.1) is:

[0031] 5'-GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCTCTCC-3'.

[0032] (2) Other conventional reagents required for reverse transcription

[0033] Including dNTP, 5×buffer, M-MLV reverse transcriptase; it can be a conventional reverse transcription reagent in the field.

[0034] 1.2 Real-time fluorescent quantitative PCR reagents

[0035] (1) Quantitative PCR primers

[0036] The sequences are:

[0037] Forward primer (SEQ ID NO.2): 5'-CAGGGCAGGGAAGGTG-3';

[0038] Reverse primer (SEQ ID NO.3): 5'-GTGCAGGGTCCGAGGT-3'.

[0039] (2) Real-time fluorescent quantitative PCR master mix

[0040] TB Green Premix Ex Taq II (Tli RNaseH Plus) from TaK...

experiment example 1

[0069] Experimental Example 1: Clinical Verification

[0070] 1. Method

[0071] Randomly select 120 cases of plasma samples of papillary thyroid carcinoma, of which 60 cases have lymph node metastasis (metastasis group), 60 cases have no lymph node metastasis (control group), extract exosomal RNA, and use the kit and method of Example 1 to detect plasma Exosomal miR-6879-5p level, and the primers in Example 1 were replaced with primers of miR-4707-3p, miR-4749-5p and miR-8058, and miR-4707-3p, miR-4707-3p, Levels of miR-4749-5p and miR-8058. Then the external reference Cel-miR-39 was used to quantify the aforementioned miRNAs.

[0072] The primer sequences of miR-4707-3p, miR-4749-5p and miR-8058 are shown in Table 3.

[0073] Table 3 Primer sequences of miR-4707-3p, miR-4749-5p and miR-8058

[0074]

[0075] The receiver operating characteristic curve and AUC were used to calculate the sensitivity and specificity of exosomal miR-6879-5p in predicting lymph node metast...

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Abstract

The invention discloses application of a reagent for detecting exosome miR-6879-5p in preparation of a thyroid cancer lymph node metastasis detection kit, and belongs to the field of cancer diagnosis. In the plasma exosome of a papillary thyroid carcinoma lymph node metastasis patient, the level of miR-6879-5p is obviously higher than that of a non-lymph node metastasis patient. According to the discovery, the reagent for detecting the exosome miR-6879-5p is used for preparing the thyroid cancer lymph node metastasis detection kit, whether a papillary thyroid cancer patient has lymph node metastasis or not can be accurately judged, and the kit has a good application prospect.

Description

technical field [0001] The invention belongs to the field of cancer diagnosis. Background technique [0002] Papillary thyroid carcinoma is the most common pathological subtype of thyroid cancer, and its prevalence has exploded worldwide in recent years. Since the onset of most papillary thyroid carcinomas is insidious, there are often no obvious positive symptoms and signs, and they are mostly found during physical examination; and when the diagnosis is made, about 30%-80% of the patients have cervical lymph node metastasis. Cervical lymph node metastasis not only increases the difficulty of surgery, but also prolongs the incision wound, increases the incidence of surgical complications, and seriously affects the prognosis of patients. [0003] At present, the existing preoperative diagnostic technology is mainly high-resolution color ultrasound of the neck combined with ultrasound-guided biopsy, and ultrasound detection is interfered by many factors, and its accuracy is g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/158C12Q2600/178Y02A50/30
Inventor 陈文杰
Owner WEST CHINA HOSPITAL SICHUAN UNIV