基于荧光猝灭探针的基因分型全自动系统。

Fluorescent quenching probes based genotyping with a fully automated system.

作者信息

Zhang Jie, Shi Changgen, Zhang Lei, Zhang Yan, Lu Qing, Wang Rongfang

机构信息

Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, China.

Shanghai R&D Center, DiaSys Diagnostic Systems (Shanghai) Co., Ltd., Shanghai, 201318, China.

出版信息

Heliyon. 2021 Apr 27;7(4):e06858. doi: 10.1016/j.heliyon.2021.e06858. eCollection 2021 Apr.

DOI:10.1016/j.heliyon.2021.e06858

PMID:33997392

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8100075/

Abstract

OBJECTIVE

The aim of the present study is to develop and validate a reliable and simple application for genotyping serum amyloid A1 ().

METHODS

The specific nested PCR was performed to amplify a product of gene. Two quenching probes (QPs) were designed for detecting two single nucleotide polymorphism (SNP) sites, rs1136743(C/T) and rs1136747(C/T) respectively for genotypes. The specific nested PCR and QPs of genotying was introduced into a fully automated genotyping system (I-densy, ARKRAY, Inc.), which enables the genotyping of from whole blood.

RESULTS

Six genotypes of (α, β, γ, αβ, αγ and βγ) could be determined by monitoring the fluorescence intensity of two QPs with melting temperature (TM) analysis. Total 121 clinical samples were genotyped in the fluorescent quenching probes based method with a fully automated I-densy system and were further sequence confirmed with a PCR direct sequencing approach.

CONCLUSION

This fully automated system is a rapid and reliable strategy for the genotyping and for its future clinical application.

摘要

目的

本研究的目的是开发并验证一种用于血清淀粉样蛋白A1（）基因分型的可靠且简单的应用方法。

方法

进行特异性巢式PCR以扩增基因的一个产物。设计了两种淬灭探针（QPs）分别用于检测两种单核苷酸多态性（SNP）位点，即rs1136743（C/T）和rs1136747（C/T）的基因型。将基因型分型的特异性巢式PCR和QPs引入全自动基因分型系统（I-densy，ARKRAY公司），该系统能够对全血中的进行基因分型。