Homologous genomic regions for characterization associated with biological targets

1. A method for facilitating characterization of a single gene disorder from cell-free nucleic acids, the method comprising:

generating a co-amplified mixture based on co-amplifying:

cell-free nucleic acids comprising:

a genomic region of interest associated with the single gene disorder; and

a homologous native genomic region with partial sequence similarity to the genomic region of interest; and

synthetic molecules, each synthetic molecule comprising:

a target-associated region matching a first portion of the genomic region of interest; and

a variation region differing from a second portion of the genomic region of interest;

sequencing the co-amplified mixture to generate first sequencing data and second sequencing data, the first sequencing data associated with the genomic region of interest and the homologous native genomic region, and the second sequencing data associated with the target-associated region and the variation region, the second sequencing data indicating a number of synthetic molecules;

determining an abundance metric for the genomic region of interest and an abundance metric for the homologous native genomic region, the abundance metric for the genomic region of interest based on the first sequencing data and second sequencing data;

and

facilitating the characterization of the single gene disorder based on the abundance metric for the genomic region of interest and the abundance metric for the homologous native genomic region.

2. The method of claim 1 ,

wherein the cell-free nucleic acids comprise first nucleic acid molecules and second nucleic acid molecules,

wherein the first nucleic acid molecules comprise the genomic region of interest,

wherein the second nucleic acid molecules comprise the homologous native genomic region, and

wherein co-amplifying a set of nucleic acid molecules comprises co-amplifying the first and the second nucleic acid molecules based on a primer pair designed based on the partial sequence similarity between the genomic region of interest and the homologous native genomic region.

3. The method of claim 2 ,

wherein the primer pair corresponds to a first primer binding sequence region and a second primer binding sequence region,

wherein the genomic region of interest and the homologous native genomic region each comprise the first primer binding sequence region and the second primer binding sequence region,

wherein the homologous native genomic region further comprises a sequence dissimilarity region with sequence dissimilarity to a corresponding sequence region of the genomic region of interest, and

wherein the sequence dissimilarity region is located between the first primer binding sequence region and the second primer binding sequence region of the homologous native genomic region.

4. The method of claim 1 , further comprising:

determining a reference ratio for the genomic region of interest relative to the homologous native genomic region, based on a number of genomic regions homologous to the genomic region of interest,

wherein facilitating the characterization of the single gene disorder comprises:

determining an observed ratio based on the abundance metric for the genomic region of interest relative to the abundance metric for the homologous native genomic region; and

comparing the observed ratio to the reference ratio.

5. The method of claim 1 , wherein the partial sequence similarity between the genomic region of interest and the homologous native genomic region satisfies a homology threshold of at least 80% homology.

6. The method of claim 1 ,

wherein the cell-free nucleic acids comprise:

a set of genomic regions of interest comprising the genomic region of interest, and

a set of homologous native genomic regions comprising the homologous native genomic regions; and

wherein facilitating the characterization of the single gene disorder comprises facilitating the characterization of the single gene disorder based on abundance metric ratios between abundance metrics for the set of genomic regions of interest and abundance metrics for the set of homologous native genomic regions.

7. The method of claim 1 ,

wherein the single gene disorder comprises a spinal muscular atrophy condition,

wherein the genomic region of interest is from SMN1 gene,

wherein the genomic region of interest comprises at least one of chr5:70238181-70238382, chr5:70247763-70248428, and chr5:70247785-70248019, and

wherein the homologous native genomic region comprises at least one of chr9:20331713-20331910, chr9:20332293-20332902, and chr4:183948110-183948725.

8. The method of claim 1 ,

wherein the single gene disorder comprises an alpha-thalassemia condition,

wherein the genomic region of interest is from HBA1 gene,

wherein the homologous native genomic region is from HBA2 gene, and

wherein facilitating the characterization of the single gene disorder comprises facilitating the characterization of the alpha-thalassemia condition based on the abundance metric for the genomic region of interest from the HBA1 gene relative to the abundance metric for the homologous native genomic region from the HBA2 gene.

9. The method of claim 1 , further comprising:

co-amplifying a set of reference-associated molecules, and second nucleic acid molecules from the cell-free nucleic acids, wherein the set of reference-associated molecules comprises:

reference-associated regions with sequence similarity to homologous native genomic region; and

reference variation regions with sequence dissimilarity to a sequence region of the homologous native genomic region; and

determining an abundance metric for the set of reference-associated molecules,

wherein facilitating the characterization of the single gene disorder comprises facilitating the characterization of the single gene disorder based on the abundance metric for the genomic region of interest, the abundance metric for the homologous native genomic region, the abundance metric for the set of synthetic molecules, and the abundance metric for the set of reference-associated molecules.

10. The method of claim 1 , wherein the abundance metric for the genomic region of interest comprises an intensity metric for the genomic region of interest, wherein the abundance metric for the homologous native genomic region comprises an intensity metric for the homologous native genomic region, and wherein determining the abundance metric for the genomic region of interest and the abundance metric for the homologous native genomic region comprises determining the intensity metric for the genomic region of interest and the intensity metric for the homologous native genomic region based on sequence dissimilarity between the genomic region of interest and the homologous native genomic region.

11. The method of claim 2 , wherein generating the co-amplified mixture comprises amplifying the genomic region of interest based on a set of primers designed for amplification of the genomic region of interest associated with at least one of HBA1 gene and HBA2 gene, the set of primers comprises primers designed for amplification of a set of genomic regions of interest corresponding to polymorphic alleles associated with the at least one of HBA1 gene and HBA2 gene, wherein the polymorphic alleles are within a SEA deletion, wherein the polymorphic alleles comprise at least one of rs2858935, rs397817350, rs1203833, rs2541675, rs2974771, rs57397665, rs3020596, rs2541669, rs2238369, rs11639532, rs2858942, rs1005364, rs11863726, rs3062451, rs28673162, and rs3859139, wherein the set of genomic regions of interest comprises the genomic region of interest, and wherein the method further comprises:

determining an allele characterization based on abundance metrics for the set of genomic regions of interest;

determining an absence or presence of a father-associated deletion corresponding to the at least one of the HBA1 gene and the HBA2 gene based on a comparison between the allele characterization and a mother allele characterization for the mother; and

facilitating the prenatal characterization of the single gene disorder based on the absence or presence of the father-associated deletion.

12. The method of claim 1 , wherein the genomic region of interest comprises a point mutation in the HBA2 gene and the synthetic molecules comprise quality control template (QCT) molecule, wherein the method further comprises:

adding, to the cell-free nucleic acids, a set of the quality control template (QCT) molecules; and

wherein facilitating the prenatal characterization of the single gene disorder comprises facilitating the characterization of the single gene disorder based on an absolute count for the genomic region of interest determined from the abundance metric for the genomic region of interest and an abundance metric for the set of QCT molecules.

13. The method of claim 1 , wherein the cell-free nucleic acids are from a maternal sample.

14. The method of claim 3 , wherein determining the abundance metric for the genomic region of interest and the abundance metric for the homologous native genomic region is based on sequence dissimilarity between the genomic region of interest and the homologous native genomic region.

15. The method of claim 1 , wherein the method further comprises facilitating a prenatal diagnosis of the single gene disorder based on the abundance metric for the genomic region of interest relative to the abundance metric for the homologous native genomic region.

16. The method of claim 7 , further comprising:

determining a second homologous native genomic region with partial sequence similarity to the genomic region of interest from the SMN1 gene,

wherein facilitating the characterization of the single gene disorder comprises facilitating the characterization of the spinal muscular atrophy condition based on the abundance metric for the genomic region of interest from the SMN1 gene, the abundance metric for the homologous native genomic region from the SMN2 gene, and an abundance metric for the second homologous native genomic region.