Development of a method for the separation of HLA-A, -B and -C haploid using biotinylated probe and streptavidin magnetic beads / 中华医学遗传学杂志
Chinese Journal of Medical Genetics
;
(6): 383-387, 2014.
Article
in Chinese
| WPRIM
| ID: wpr-291766
ABSTRACT
<p><b>OBJECTIVE</b>To develop a method for separating the human leukocyte antigen (HLA)-A, -B and -C haploid using biotinylated probes and streptavidin magnetic beads in order to solve ambiguous HLA genotyping results.</p><p><b>METHODS</b>Based on sequence information of HLA alleles from the IMGT/HLA database, the 5-biotinylated probes were designed. The probe was mixed and extended with corresponding genomic DNA, and incubated with streptavidin magnetic beads, which could form a streptavidin magnetic beads-biotin-probe DNA complex. The unique DNA haploid binding to corresponding probe was isolated after washes and elution. The separated haploid genomic DNA was used as template for HLA-A, -B and -C loci amplification and sequencing analysis.</p><p><b>RESULTS</b>Among the 12 HLA-A probes, 19 HLA-B probes and 13 HLA-C probes, DNA sequencing has confirmed that 9 HLA-A probes, 9 HLA-B probes and 5 HLA-C probes could successfully separate the haploid from genomic DNA samples.</p><p><b>CONCLUSION</b>The developed method for HLA-A, -B and -C haploid separation is reliable, which can solve certain ambiguity and improve the accuracy of HLA genotyping.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
HLA-A Antigens
/
HLA-B Antigens
/
HLA-C Antigens
/
Chemistry
/
Polymerase Chain Reaction
/
Molecular Probe Techniques
/
Streptavidin
/
Genetics
/
Genotype
/
Haploidy
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Medical Genetics
Year:
2014
Type:
Article
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