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Application of hsa-miR-378i as marker molecule to preparation of tuberculosis diagnosis reagent kit and detection reagent kit for tuberculosis diagnosis

A technology of hsa-mir-378i and detection reagent, applied in the field of biomedicine, can solve the problems of low detection sensitivity, inability to achieve high sensitivity and high specificity, etc., and achieve the effect of great application prospect, market prospect and high specificity

Pending Publication Date: 2020-01-17
WUXI NO 2 PEOPLES HOSPITAL
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

The prior art discloses a technical solution for diagnosing tuberculosis by using miRNA in plasma exosomes as a marker molecule. However, its detection sensitivity is low, and high sensitivity and high specificity detection standards cannot be achieved.

Method used

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  • Application of hsa-miR-378i as marker molecule to preparation of tuberculosis diagnosis reagent kit and detection reagent kit for tuberculosis diagnosis
  • Application of hsa-miR-378i as marker molecule to preparation of tuberculosis diagnosis reagent kit and detection reagent kit for tuberculosis diagnosis
  • Application of hsa-miR-378i as marker molecule to preparation of tuberculosis diagnosis reagent kit and detection reagent kit for tuberculosis diagnosis

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Embodiment 1

[0100] A total of 65 patients with Mycobacterium tuberculosis infection and 65 healthy controls were selected. The subjects fasted for 12 hours. The next morning, 5 mL of cubital median venous blood was collected with a blood collection tube containing sodium citrate anticoagulant. Centrifuge at 3000r / min for 10min, and it can be seen that it is divided into two layers. Use a pipette gun to absorb the transparent light yellow liquid in the upper layer, that is, plasma. Plasma exosomes were extracted using the Invitrogen exosome extraction kit from Life. Use Trizol to extract total RNA in exosomes, reverse transcribe into cDNA, use TruSeq Small RNA Sample Prep Kits to construct a library, and perform IlluminaHiSeq high-throughput sequencing on the resulting library, using SE50 sequencing strategy to remove low-quality sequences, 3' adapters, 5' contamination, poly A fragments, and <18nt sequences to obtain high-quality small RNA sequences. The number, type and length distribut...

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Abstract

The invention discloses an application of hsa-miR-378i as a marker molecule to preparation of a tuberculosis diagnosis reagent kit and a detection reagent kit for tuberculosis diagnosis. The hsa-miR-378i is from a plasma exosome, and the base sequence of the hsa-miR-378i is shown as SEQID NO:1. The detection reagent kit contains a detection reagent for specially detecting the marker molecule. Besides, the invention further discloses an application to preparation the tuberculosis diagnosis reagent kit through union of hsa-miR-378i and one or two of hsa-miR-148a-3p and hsa-miR-150-5p as the marker molecule. Experimental result indicates that hsa-miR-378i has high specificity, sensitivity and diagnosis effects on tuberculosis diagnosis. Therefore, the application has great application prospects and market prospects to the field of tuberculosis diagnosis.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to the use of hsa-miR-378i as a marker molecule in preparing a diagnostic kit for tuberculosis and a detection kit thereof. Background technique [0002] Currently, tuberculosis (TB) has become an important cause of death worldwide, largely due to difficulties in diagnosis and treatment of TB patients. Early detection and early diagnosis are the key links to reduce the chance of transmission and control the tuberculosis epidemic. Therefore, the identification of new biomarkers for timely and accurate diagnosis of tuberculosis plays a decisive role in the control of tuberculosis. Studies in recent years have shown that exosomes are closely related to Mycobacterium tuberculosis infection. Exosomes are extracellular vesicles secreted by a variety of living cells, with a size of about 30-100nm, containing a variety of biologically active components, and participating in ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/158C12Q2600/178C12Q2600/166
Inventor 尹斌张敏
Owner WUXI NO 2 PEOPLES HOSPITAL
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