Preparation of robust synthetic control samples and their use in a metatranscriptomic clinical test

Abstract Metatranscriptomics (MT) has the potential to revolutionize the field of molecular diagnostics. Due to the complexity of MT diagnostic models, positive and negative control materials for specific disease indications can be difficult to obtain. Controls must often be sourced directly from pa...

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Main Authors: Ryan Toma, Lan Hu, Guru Banavar, Momchilo Vuyisich
Format: Article
Language:English
Published: Nature Portfolio 2025-03-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-025-95020-y
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author Ryan Toma
Lan Hu
Guru Banavar
Momchilo Vuyisich
author_facet Ryan Toma
Lan Hu
Guru Banavar
Momchilo Vuyisich
author_sort Ryan Toma
collection DOAJ
description Abstract Metatranscriptomics (MT) has the potential to revolutionize the field of molecular diagnostics. Due to the complexity of MT diagnostic models, positive and negative control materials for specific disease indications can be difficult to obtain. Controls must often be sourced directly from patients. This introduces logistical burdens, assay variability, and limits high throughput clinical laboratory operations. To overcome this limitation, we developed a method for generating Synthetic Control (SC) samples, which duplicate the nucleic acid signature of complex clinical specimens and produce the desired test outcome. SCs can be easily and cost-effectively produced in large quantities (> 100,000 SCs per amplification cycle), enabling high throughput diagnostic testing. Here, we report the generation of Synthetic Positive Control (SPC) samples. SPCs were validated and implemented in a clinical laboratory. The SPCs produced robust positive signals (average OC risk score of 0.996) and high levels of reproducibility (%CV of 0.29%) in a high throughput automated CLIA laboratory. SCs are a novel and useful method for the generation of high quality controls for MT-based diagnostic tests, and their adoption could herald the widespread use of MT tests in molecular diagnostics.
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spelling doaj-art-ec93eebfb28b44828ea215abe84162302025-08-20T02:49:23ZengNature PortfolioScientific Reports2045-23222025-03-011511810.1038/s41598-025-95020-yPreparation of robust synthetic control samples and their use in a metatranscriptomic clinical testRyan Toma0Lan Hu1Guru Banavar2Momchilo Vuyisich3Viome Research Institute, Viome Life Sciences, Inc.Viome Research Institute, Viome Life Sciences, Inc.Viome Research Institute, Viome Life Sciences, Inc.Viome Research Institute, Viome Life Sciences, Inc.Abstract Metatranscriptomics (MT) has the potential to revolutionize the field of molecular diagnostics. Due to the complexity of MT diagnostic models, positive and negative control materials for specific disease indications can be difficult to obtain. Controls must often be sourced directly from patients. This introduces logistical burdens, assay variability, and limits high throughput clinical laboratory operations. To overcome this limitation, we developed a method for generating Synthetic Control (SC) samples, which duplicate the nucleic acid signature of complex clinical specimens and produce the desired test outcome. SCs can be easily and cost-effectively produced in large quantities (> 100,000 SCs per amplification cycle), enabling high throughput diagnostic testing. Here, we report the generation of Synthetic Positive Control (SPC) samples. SPCs were validated and implemented in a clinical laboratory. The SPCs produced robust positive signals (average OC risk score of 0.996) and high levels of reproducibility (%CV of 0.29%) in a high throughput automated CLIA laboratory. SCs are a novel and useful method for the generation of high quality controls for MT-based diagnostic tests, and their adoption could herald the widespread use of MT tests in molecular diagnostics.https://doi.org/10.1038/s41598-025-95020-y
spellingShingle Ryan Toma
Lan Hu
Guru Banavar
Momchilo Vuyisich
Preparation of robust synthetic control samples and their use in a metatranscriptomic clinical test
Scientific Reports
title Preparation of robust synthetic control samples and their use in a metatranscriptomic clinical test
title_full Preparation of robust synthetic control samples and their use in a metatranscriptomic clinical test
title_fullStr Preparation of robust synthetic control samples and their use in a metatranscriptomic clinical test
title_full_unstemmed Preparation of robust synthetic control samples and their use in a metatranscriptomic clinical test
title_short Preparation of robust synthetic control samples and their use in a metatranscriptomic clinical test
title_sort preparation of robust synthetic control samples and their use in a metatranscriptomic clinical test
url https://doi.org/10.1038/s41598-025-95020-y
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AT lanhu preparationofrobustsyntheticcontrolsamplesandtheiruseinametatranscriptomicclinicaltest
AT gurubanavar preparationofrobustsyntheticcontrolsamplesandtheiruseinametatranscriptomicclinicaltest
AT momchilovuyisich preparationofrobustsyntheticcontrolsamplesandtheiruseinametatranscriptomicclinicaltest