Haplotype Region Handling

In this document, we will go into further detail about how regions are reported in the alternative allele field of an hVCF file and possible edge cases that may arise in hVCF creation.

Alternative allele fields - an overview

Note

For more information about this hVCF field and other general information about hVCF specifications, please review the haplotype VCF specification article.

The alternative allele (##ALT) field contains key-value information that represents the haplotype sequence and ID for a given reference-range in the PHG. One key-value pair is the Regions key. This represents the physical location of the sequence in the genome assembly FASTA file.

A common example of this would be the following example:

##ALT=<ID=57705b1e2541c7634ea59a48fc52026f,Description="haplotype data for line: LineA",Source="data/test/smallseq/LineA.fa",Regions=1:14-19,Checksum=Md5,RefRange=06ae4e937668d301e325d43725a38c3f>

...we have the following regions key:

Regions=1:14-19

...for the following assembly/sample:

Description="haplotype data for line: LineA"

This indicates that for LineA, we have a haplotype sequence that aligned against the given reference range (denoted with the MD5 hash, 06ae4e937668d301e325d43725a38c3f) at chromosome 1 (1:) between the values of 14 and 19 (14-19) base-pairs. We can represent this graphically with a dot plot:

In the above example, LineA (y-axis) is aligned against our reference genome, in this case, identified as Reference (x-axis). The reference range boundary is located on chromosome 1 between the base pairs of 3 and 8 (denoted by the dashed lines). Given our hypothetical alignment scenario, our sample, LineA, aligned against the reference genome also on chromosome 1, but due to structural variations between the two genomes, matches at different base-pair positions (in this case between 14 and 19 base-pairs). Using this region value, we can navigate back to LineA's FASTA file and identify the sequence region:

Possible edge cases - inversions

Given the possibility of structural diversity events between the reference and assembly genomes. There are edge cases where a reference range boundary can land on inversion regions. For example, let's illustrate this scenario with another dot plot:

Here, when we align LineA against the reference, there is an inversion at the end of this boundary. How do we represent this using our Region key-value pairing scheme in the alternate allele field? The Region key can house multiple region elements using comma (,) separated values. Using the above example, this can be displayed as follows:

Regions="1:13-16,1:21-19"

In the above example, we provide multiple regions (separated by commas) to indicate the normal and inverted portions of the alignment:

Region	Chromosome	Range (bp)	Type
1:13-16	1	13 to 16	normal
1:21-19	1	21 to 19	inverted

Since the last element of the region is inverted, the larger value will be the first value left of the - character and the smaller value will be to the right of the - character. This will indicate to PHGv2 that reverse transcription of the sequence will need to occur before the MD5 hashing step.