Allele-specific electrical genotyping for diagnosis of transthyretin amyloidosis

Abstract Clinical genetic testing often takes days to weeks, but rapid, affordable tests during outpatient visits could significantly benefit patients. This is crucial for detecting common, actionable point mutations, such as those linked to hereditary transthyretin (TTR) amyloidosis, which is often...

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Main Authors: Muhammad Tayyab, Neeru Gandotra, Jianye Sui, Curt Scharfe, Mehdi Javanmard
Format: Article
Language:English
Published: Nature Portfolio 2025-03-01
Series:Communications Engineering
Online Access:https://doi.org/10.1038/s44172-025-00385-7
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author Muhammad Tayyab
Neeru Gandotra
Jianye Sui
Curt Scharfe
Mehdi Javanmard
author_facet Muhammad Tayyab
Neeru Gandotra
Jianye Sui
Curt Scharfe
Mehdi Javanmard
author_sort Muhammad Tayyab
collection DOAJ
description Abstract Clinical genetic testing often takes days to weeks, but rapid, affordable tests during outpatient visits could significantly benefit patients. This is crucial for detecting common, actionable point mutations, such as those linked to hereditary transthyretin (TTR) amyloidosis, which is often underdiagnosed in individuals of West African ancestry with congestive heart failure. Here we developed a method for detecting known DNA variants using allele-specific polymerase chain reaction (ASPCR) and electrical impedance. Oligonucleotide primers were designed to selectively amplify the ancestral and variant allele of TTR c.424G>A (p.Val142Ile). PCR products were detected in solution using passive-flow microfluidic impedance cytometry across multiple excitation frequencies. Impedance responses correlated with DNA concentration, allowing for the calculation of a robust DNA quantification score. TTR V142I genotypes from six patients (four heterozygous, two wildtype) matched results from gel electrophoresis and sequencing. Future improvements will focus on reducing DNA input requirements and enabling multiplex variant detection.
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spelling doaj-art-3090f894989b449fa8f91a0e2d6e9b3c2025-08-20T02:56:20ZengNature PortfolioCommunications Engineering2731-33952025-03-01411810.1038/s44172-025-00385-7Allele-specific electrical genotyping for diagnosis of transthyretin amyloidosisMuhammad Tayyab0Neeru Gandotra1Jianye Sui2Curt Scharfe3Mehdi Javanmard4Department of Electrical and Computer Engineering, Rutgers UniversityDepartment of Genetics, Yale University School of MedicineDepartment of Electrical and Computer Engineering, Rutgers UniversityDepartment of Genetics, Yale University School of MedicineDepartment of Electrical and Computer Engineering, Rutgers UniversityAbstract Clinical genetic testing often takes days to weeks, but rapid, affordable tests during outpatient visits could significantly benefit patients. This is crucial for detecting common, actionable point mutations, such as those linked to hereditary transthyretin (TTR) amyloidosis, which is often underdiagnosed in individuals of West African ancestry with congestive heart failure. Here we developed a method for detecting known DNA variants using allele-specific polymerase chain reaction (ASPCR) and electrical impedance. Oligonucleotide primers were designed to selectively amplify the ancestral and variant allele of TTR c.424G>A (p.Val142Ile). PCR products were detected in solution using passive-flow microfluidic impedance cytometry across multiple excitation frequencies. Impedance responses correlated with DNA concentration, allowing for the calculation of a robust DNA quantification score. TTR V142I genotypes from six patients (four heterozygous, two wildtype) matched results from gel electrophoresis and sequencing. Future improvements will focus on reducing DNA input requirements and enabling multiplex variant detection.https://doi.org/10.1038/s44172-025-00385-7
spellingShingle Muhammad Tayyab
Neeru Gandotra
Jianye Sui
Curt Scharfe
Mehdi Javanmard
Allele-specific electrical genotyping for diagnosis of transthyretin amyloidosis
Communications Engineering
title Allele-specific electrical genotyping for diagnosis of transthyretin amyloidosis
title_full Allele-specific electrical genotyping for diagnosis of transthyretin amyloidosis
title_fullStr Allele-specific electrical genotyping for diagnosis of transthyretin amyloidosis
title_full_unstemmed Allele-specific electrical genotyping for diagnosis of transthyretin amyloidosis
title_short Allele-specific electrical genotyping for diagnosis of transthyretin amyloidosis
title_sort allele specific electrical genotyping for diagnosis of transthyretin amyloidosis
url https://doi.org/10.1038/s44172-025-00385-7
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