A pilot study of the performance of NeoEXOME for NBS in China
To evaluate the performance of NeoEXOME in clinical practice. A pilot, multi-center clinical trial was conducted. From Oct 2019 to Sep 2021, 3423 neonates were enrolled from 5 institutions (Fig 2). Of all these subjects, DBS was collected within 3 days of birth and next-generation sequencing was performed based on our NeoEXOME panel.
Among the 3249 neonates eligible for the analysis, 934 infants were NGS negative, and 389 infants were NGS positive (244 High risk, 4 Moderate risk, 141 Low risk), and 1926 were carriers. NGS positive rate was 12.0%. 343 infants showed one genetic mutation, who were genetically susceptible to mono gene-related diseases; 46 infants had more than one gene mutations, who were genetically susceptible to multi-diseases. Infants that were NGS reported as high-risk account for the highest proportion (Fig 3A-B). While autosomal recessive inheritance was the most common inheritance pattern (Fig 3A-B). Endocrine and metabolism system disorders accounted for the highest proportion in healthy neonates (Fig 3C). Most of the diseases detected were predicted to develop within the first year of life (Fig 3D). Variants of DUOX2account for the most frequent mutant genes (18.3%, 131/716), followed by UGT1A1 , PAH , GJB2 , FLG (Fig 3E). The top five related disorders were thyroid dyshormonogenesis, hyperbilirubinemia, phenylketonurics, deafness, hyperbilirubinemia, and ichthyosis vulgaris, with the frequency of variant 5.7%, 5.4%, 5.0 %, 4.1%, respectively. 1926 infants were tested as carrier of the diseases in our panel. There were 3462 variants, including 1224 pathogenic variants and 2238 likely pathogenic variants (Fig 4A). Variants ofGJB2 account for the most frequent mutant genes of all the centers, followed by UGT1A1 , DUOX2, FLG, andSLC25A13 (Fig 4B).
To further validate the use of NGS in NBS, conventional NBS results were collected to compare the consistency of both approaches. 200 out of 3249 neonates were conventionally NBS positive (+). In the NBS (+) subgroup, 118 were NGS positive (64 High risk, 2 Moderate risk, 52 Low risk), 14 were NGS negative, and 68 were NGS carrier (Table 2). Of the 118 NBS (+)/NGS (+) cases, 90 (76.3%) had consistent results with conventional NBS and NGS (Supplementary Table 3). In the 3049 NBS (-) subgroup, 271 (8.9%) were NGS positive (180 High risk, 2 Moderate risk, 89 Low risk), 920 were NGS negative, and 1858 were NGS carrier (Table 2). 168 of the 271 NGS (+) /NBS (-) neonates were followed up till Dec 2021, and 9 of them (including genes of DUXO2 , PAH , MUT , WAS , and SLC22A5 ) were clinically diagnosed.