Novel Modification of a Confirmatory SMA Sequencing Assay that Can Be Used to Determine SMN2 Copy Number
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. DNA Isolation from Dry Blood Spot
2.3. DNA Primers and Sequencing
2.4. Validation of SMN1 Sequencing Assay
2.5. Modification of Sequencing Assay for Determination of SMN2 Copy Number
3. Results
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Type of Reference Specimens | Number of Specimens | NENSP Detection of Nucleotide at c.840 of SMN1 |
---|---|---|
Affected | n = 14 | “T” |
Parents | n = 16 | “C” and “T” |
Carrier | “C” and “T” | |
Positive control (known SMA) | “T” |
SMN2 Copy Number Assay Applied to the Specimens of Infants Identified by NBS as SMA Affected. | |||
---|---|---|---|
MA Patient Specimens | SMA Status | SMN2 Copy Number Prediction by NENSP from DBS | SMN2 Copy Number Prediction by Diagnostic Lab Test of an Independent Specimen |
TS1 | Affected | 3 | 2 |
TS2 | Affected | More than 3 copies | 4 |
TS3 | Affected | 2 | 2 |
TS4 | Affected | 3 | 2 |
TS5 | Affected | 2 | 2 |
TS6 | Affected | 2 | 2 |
TS7 | Affected | 4 | 4 |
TS8 | Affected | 3 | 2 |
TS9 | Affected | 2 | 2 |
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Kumar, B.; Barton, S.; Kordowska, J.; Eaton, R.B.; Counihan, A.M.; Hale, J.E.; Comeau, A.M. Novel Modification of a Confirmatory SMA Sequencing Assay that Can Be Used to Determine SMN2 Copy Number. Int. J. Neonatal Screen. 2021, 7, 47. https://doi.org/10.3390/ijns7030047
Kumar B, Barton S, Kordowska J, Eaton RB, Counihan AM, Hale JE, Comeau AM. Novel Modification of a Confirmatory SMA Sequencing Assay that Can Be Used to Determine SMN2 Copy Number. International Journal of Neonatal Screening. 2021; 7(3):47. https://doi.org/10.3390/ijns7030047
Chicago/Turabian StyleKumar, Binod, Samantha Barton, Jolanta Kordowska, Roger B. Eaton, Anne M. Counihan, Jaime E. Hale, and Anne Marie Comeau. 2021. "Novel Modification of a Confirmatory SMA Sequencing Assay that Can Be Used to Determine SMN2 Copy Number" International Journal of Neonatal Screening 7, no. 3: 47. https://doi.org/10.3390/ijns7030047