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Chinese Journal of Medical Genetics ; (6): 383-387, 2014.
Artículo en Chino | WPRIM | ID: wpr-291766

RESUMEN

<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>


Asunto(s)
Humanos , Genotipo , Antígenos HLA-A , Genética , Antígenos HLA-B , Genética , Antígenos HLA-C , Genética , Haploidia , Técnicas de Sonda Molecular , Reacción en Cadena de la Polimerasa , Métodos , Estreptavidina , Química
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