From saliva to SNP: non-invasive, point-of-care genotyping for precision medicine applications using recombinase polymerase amplification and giant magnetoresistive nanosensors,Lab on a Chip

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From saliva to SNP: non-invasive, point-of-care genotyping for precision medicine applications using recombinase polymerase amplification and giant magnetoresistive nanosensors
Lab on a Chip ( IF 6.1 ) Pub Date : 2022-05-07 , DOI: 10.1039/d2lc00233g
Ana Sofia de Olazarra ₁ , Dana Lee Cortade ₂ , Shan X Wang _{1,

2}

Affiliation

Genetic testing is considered a cornerstone of the precision medicine paradigm. Genotyping of single nucleotide polymorphisms (SNPs) has been shown to provide insights into several important issues, including therapy selection and drug responsiveness. However, a scarcity of widely deployable and cost-effective genotyping tools has limited the integration of precision medicine into routine clinical practice. The objective of our work was to develop a portable, cost-effective, and automated platform that performs SNP genotyping at the point-of-care (POC). Using recombinase polymerase amplification (RPA) and giant magnetoresistive (GMR) nanosensors, we present a highly automated and multiplexed point-of-care platform that utilizes direct saliva for the qualitative genotyping of four SNPs (rs4633, rs4680, rs4818, rs6269) along the catechol-O-methyltransferase gene (COMT), which is associated with the modulation of pain sensitivity and perioperative opioid use. Using this approach, we successfully amplify, detect, and genotype all four of the SNPs, demonstrating 100% accordance between the experimental results obtained using the automated RPA and GMR genotyping assay and the results obtained using a COMT PCR genotyping assay that was formerly validated using pyrosequencing. This automated, portable, and multiplexed RPA and GMR assay shows great promise as a solution for SNP genotyping at the POC and reinforces the broad applications of magnetic nanotechnology in biomedicine.

中文翻译：

从唾液到 SNP：使用重组酶聚合酶扩增和巨磁阻纳米传感器进行精准医疗应用的非侵入性即时基因分型

基因检测被认为是精准医学范式的基石。单核苷酸多态性 (SNP) 的基因分型已被证明可以提供对几个重要问题的见解，包括治疗选择和药物反应性。然而，可广泛部署且具有成本效益的基因分型工具的稀缺性限制了精准医学与常规临床实践的整合。我们工作的目标是开发一个便携式、具有成本效益的自动化平台，在床旁 (POC) 执行 SNP 基因分型。使用重组酶聚合酶放大 (RPA) 和巨磁阻 (GMR) 纳米传感器，我们提出了一个高度自动化和多路复用的即时平台，该平台利用直接唾液对四个 SNP（rs4633、rs4680、rs4818、rs6269）进行定性基因分型。儿茶酚-O-甲基转移酶基因 ( COMT )，与疼痛敏感性的调节和围手术期阿片类药物的使用有关。使用这种方法，我们成功地扩增、检测和基因分型所有四个 SNP，证明使用自动化 RPA 和 GMR 基因分型测定获得的实验结果与使用以前使用验证的COMT PCR 基因分型测定获得的结果之间 100% 一致焦磷酸测序。这种自动化、便携式和多路复用的 RPA 和 GMR 检测作为 POC 的 SNP 基因分型解决方案显示出巨大的前景，并加强了磁性纳米技术在生物医学中的广泛应用。

更新日期：2022-05-10

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