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开发用于定量检测流行和具有生物医学意义的人类肠道微生物类群的 qPCR 平台。

Development of qPCR platform with probes for quantifying prevalent and biomedically relevant human gut microbial taxa.

机构信息

Knomics LLC, Skolkovo Innovation Center, Bolshoy Bulvar str., building 42, premise 1, room 1639, Moscow, 143026, Russia; Atlas Biomed Group, 31 Malaya Nikitskaya str., 121069, Moscow, Russia.

Knomics LLC, Skolkovo Innovation Center, Bolshoy Bulvar str., building 42, premise 1, room 1639, Moscow, 143026, Russia; Atlas Biomed Group, 31 Malaya Nikitskaya str., 121069, Moscow, Russia.

出版信息

Mol Cell Probes. 2020 Aug;52:101570. doi: 10.1016/j.mcp.2020.101570. Epub 2020 Apr 15.

Abstract

Nowadays the advent of innovative high-throughput sequencing allows obtaining high-quality microbiome profiling. However, PCR-based tests are still considered the "golden standard" for many clinical applications. Here, we designed a qPCR-based platform with fluorescent-labeled oligonucleotide probes for assessing human gut microbiome composition. The system allows conducting qualitative and semiquantitative analysis for 12 prokaryotic taxa that are prevalent in the human gut and associated with diseases, diet, age and other factors. The platform was validated by comparing microbiome profile data obtained with two different methods - the platform and high-throughput 16S rRNA sequencing - across 42 stool samples. The test can form the basis for precise and cost-efficient microbiome assay for large-scale surveys including clinical trials with interventions related to diet and disease risks.

摘要

如今,创新的高通量测序技术的出现使得获得高质量的微生物组谱成为可能。然而,基于 PCR 的检测仍然被认为是许多临床应用的“金标准”。在这里,我们设计了一种基于 qPCR 的荧光标记寡核苷酸探针平台,用于评估人类肠道微生物组的组成。该系统允许对人类肠道中常见的 12 种与疾病、饮食、年龄和其他因素相关的原核生物进行定性和半定量分析。该平台通过比较两种不同方法(平台和高通量 16S rRNA 测序)在 42 个粪便样本中获得的微生物组谱数据进行了验证。该检测方法可以为包括与饮食和疾病风险相关的干预措施的临床试验在内的大规模调查提供精确和具有成本效益的微生物组分析基础。

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