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Application of single cell sequencing as marker in preparation of diagnostic primary sicca syndrome

A technology for Sjögren's syndrome and single-cell sequencing, which is applied in biochemical equipment and methods, and the determination/examination of microorganisms to achieve the effects of reduced pain, high sensitivity, and easy sequencing.

Pending Publication Date: 2021-10-29
FIRST AFFILIATED HOSPITAL OF ANHUI UNIV OF CHINESE MEDICINE
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Problems solved by technology

Single-cell sequencing has not been applied to the diagnosis of Sjögren's syndrome

Method used

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  • Application of single cell sequencing as marker in preparation of diagnostic primary sicca syndrome
  • Application of single cell sequencing as marker in preparation of diagnostic primary sicca syndrome
  • Application of single cell sequencing as marker in preparation of diagnostic primary sicca syndrome

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Embodiment Construction

[0023] The invention provides an application of single-cell sequencing as a marker in preparing and diagnosing primary Sjögren's syndrome.

[0024] Peripheral blood mononuclear cells (PBMC) were isolated from controls (CN) and active primary Sjögren's syndrome (pSS) for single-cell RNA sequencing (scRNA-seq) using the 10x Genomics platform. Unsupervised clustering of cells based on gene expression profiles was performed using the Seurat package and passed to UMAP for cluster visualization (e.g. figure 1 ).

[0025] specifically,

[0026] 1. Single-cell RNA-seq workflow

[0027] PBMCs were isolated from whole blood. Single-cell whole-transcriptome analysis was performed based on the BD-Rhapsody system whole-transcriptome analysis alpha protocol, and the captured single-cell transcriptome microbeads were constructed into a cDNA library containing cell label and UMI information. Briefly, double-stranded cDNA is first extracted from captured single-cell transcripts, followed b...

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Abstract

The invention provides application of single cell sequencing as a marker in preparation of a diagnostic primary sicca syndrome. The single cell sequencing is obtained by separating peripheral blood mononuclear cells through flow-type activated cells, and the primary sicca syndrome is mainly composed of NK cells, and then T cells and B cells. According to the single cell sequencing technology, a genome, a transcriptome, a proteome and an epigenome are sequenced on the single cell level, heterogeneity information among different cells can be obtained from a tissue sample, the advantages of being accurate, efficient, safe and reliable are achieved, and therefore the differential expression marker of the sicca syndrome is screened out on the cell level, and the defects of sicca syndrome diagnosis are overcome. The single cell sequencing is easy and convenient, special preparation and specific time are not needed, and a specimen can be collected anytime and anywhere. In addition, the advantages of being extremely small in trauma and high in sensitivity are achieved.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and specifically relates to the application of single-cell sequencing as a marker in the preparation and diagnosis of primary Sjögren's syndrome. Background technique [0002] The onset of primary Sjogren's syndrome (pSS) is often insidious, and it is difficult for most patients to tell the exact onset time. The clinical manifestations are diverse, and multiple systems of the body may be involved. The severity of the disease varies greatly, and the treatment options are limited. pSS is clinically extensive and complex, and it is a disease that is easily misdiagnosed and missed. The formulation of its diagnostic criteria has been developed and evolved for more than 50 years, and 13 classification criteria have appeared successively. However, there are still various clinical applications. dissatisfied. pSS lacks a single and objective diagnostic index, and traditional diagnosis often ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/6869
CPCC12Q1/6883C12Q1/6869C12Q2600/158C12Q2535/122C12Q2537/165
Inventor 万磊刘健黄传兵谌曦范海霞葛瑶刘天阳刘磊李明赵磊孙广瀚朱子衡李舒马熙檬
Owner FIRST AFFILIATED HOSPITAL OF ANHUI UNIV OF CHINESE MEDICINE
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