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.
Artículo
en Chino
| 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>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Antígenos HLA-A
/
Antígenos HLA-B
/
Antígenos HLA-C
/
Química
/
Reacción en Cadena de la Polimerasa
/
Técnicas de Sonda Molecular
/
Estreptavidina
/
Genética
/
Genotipo
/
Haploidia
Límite:
Humanos
Idioma:
Chino
Revista:
Chinese Journal of Medical Genetics
Año:
2014
Tipo del documento:
Artículo
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