# Feasibility and validation of an integrated newborn screening algorithm with targeted Next Generation Sequencing (tNGS) technology as part of a 2nd-tier test for Pompe and MPS I

> **NIH NIH R44** · BAEBIES, INC. · 2021 · $793,462

## Abstract

ABSTRACT
Feasibility and validation of an integrated newborn screening algorithm with targeted Next Generation
 Sequencing (tNGS) technology as part of a 2nd-tier test for Pompe and MPS I
Newborn screening (NBS) utilizes high throughput primary (1st-tier) screening assays paired with referral and
clinical follow-up testing to identify babies who are affected with certain inherited disorders. Where necessary,
2nd-tier tests are performed prior to referral for follow-up testing in order to reduce the number of false positives
(screen positive samples that are determined to be unaffected). False-positive newborn screens have undesired
consequences for both families and the public health lab referral system, including: high cost associated with
additional confirmatory testing, extra testing burden on referral centers, parental anxiety, parent-baby bonding
issues, and added stress to the baby with additional tests and blood draws. There exists a strong need to reduce
the rate of false positive newborn screens by implementing 2nd-tier molecular or biochemical tests prior to referral.
This Phase II project will be a continuation of our successful Phase I research, which generated an integrated
2nd-tier targeted next generation sequencing (tNGS) workflow capable of identifying both point mutations and
large deletions/duplication events from dried blood spot (DBS) specimens. We will continue to focus on Pompe
disease and Mucopolysaccharidosis Type I (MPS I) -- two lysosomal storage disorders that were recently
recommended for universal NBS in the U.S., but have been challenging to implement as NBS tests due to the
high rates of pseudodeficient variants. Currently, 2nd-tier testing via either additional biochemical analysis or gene
sequencing for known pathogenic variants are used to identify pseudodeficiency and reduce false positive test
rates. We will expand our novel tNGS 2nd-tier workflow by: 1) developing bioinformatic tools for variants of
uncertain significance (VUS) cut-off and cross-reactive immunological material (CRIM) status prediction; 2)
improving our existing copy number variability (CNV) caller; and 3) integrating additional enzyme measurements
and demographic data with the tNGS score. Demographic data has previously been shown to correlate to
measured enzyme activities due to biological factors and DBS sample variability. Our algorithm will provide a
better disease state call and associated data for improved follow-up care, provide critical predictions for disease
onset and treatment considerations.
The 2nd-tier tests developed through this work will be sold initially as a diagnostic send-out service and eventually
as kits to public health labs that are currently screening, or planning to screen for Pompe disease and MPS I.
Affected individuals who are identified using our tests will be referred to follow-up earlier and will have an
accelerated path to disease confirmation and treatment. These features are especially important for Pompe
d...

## Key facts

- **NIH application ID:** 10082458
- **Project number:** 5R44HD094543-03
- **Recipient organization:** BAEBIES, INC.
- **Principal Investigator:** Viren R Amin
- **Activity code:** R44 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $793,462
- **Award type:** 5
- **Project period:** 2018-08-16 → 2022-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10082458

## Citation

> US National Institutes of Health, RePORTER application 10082458, Feasibility and validation of an integrated newborn screening algorithm with targeted Next Generation Sequencing (tNGS) technology as part of a 2nd-tier test for Pompe and MPS I (5R44HD094543-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10082458. Licensed CC0.

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