Primers and methods for determining RhD zygosity

摘要

Provided herein are primers comprising a nucleotide sequence complementary to a portion of a RhD gene. Also provided herein are methods of determining a RhD zygosity in a subject. Also provided are methods of detecting a weak D allele in a subject. Further provided are kits for determining an RhD zygosity.

主权项

1. A method of determining Rhesus D (RhD) zygosity in a subject, the method comprising:
(a) obtaining a nucleic acid comprising a RhD gene from a biological sample of the subject; (b) performing a PCR assay on the nucleic acid with a primer pair comprising
(i) a first primer comprising a nucleotide sequence with at least 93% identity to SEQ ID NO:1, wherein the primer is 20 to 31 nucleotides in length, and(ii) a second primer, wherein said second primer hybridizes under stringent conditions to a nucleotide sequence in the region between positions 2940 and 3057 of SEQ ID NO:4, SEQ ID NO:5 or SEQ ID NO:6, wherein the PCR assay results in a PCR product; (c) digesting the PCR product with a restriction enzyme; and (d) analyzing the digested PCR product for nucleotide fragments of a first, second or third pattern; wherein the digested PCR product pattern determines the RhD zygosity of the subject.