Development of a high-throughput sequence-based typing assay for human leukocyte antigen loci / 中华医学遗传学杂志
Chinese Journal of Medical Genetics
;
(6): 323-327, 2012.
Artículo
en Chino
| WPRIM
| ID: wpr-295484
ABSTRACT
<p><b>OBJECTIVE</b>To develop a reliable assay for simultaneous sequence-based typing (SBT) of HLA-DPA1 and HLA-DPB1, and to apply it for the study of allelic polymorphisms in southern Chinese Han population.</p><p><b>METHODS</b>Based on full-length HLA-DPA1 and HLA-DPB1 allelic sequences, locus-specific PCR primers were designed and applied to amplify the target sequence encompassing the entire exon 2 of HLA-DPA1 and HLA-DPB1. PCR products were purified with magnetic beads, and run through an ABI 3730 DNA sequencer. Genotypes were assigned with an Assign 3.5 SBT software.</p><p><b>RESULTS</b>The target sequences of HLA-DPA1 and HLA-DPB1 were both amplified with the PCR procedure. Little background and noise was observed in the derived sequences. Among 176 non-related healthy individuals, 4 HLA-DPA1 alleles with the frequencies of DPA1*0202 (0.589) > DPA1*0103 (0.284) > DPA1*0201 (0.096) > DPA1*0401 (0.031) were identified. In addition, 14 HLA-DPB1 alleles, including 4 common alleles (with a frequency of more than 5%, namely DPB1*0501, DPB1*0201, DPB1*0401 and DPB1*0202), 7 alleles with a frequency ranging from 1%-5% and 3 alleles with a frequency of less than 1% were identified. The results of HLA-DPB1 genotyping were all in accordance with the typing results derived from an Atria AlleleSEQR HLA-DPB1 kit.</p><p><b>CONCLUSION</b>A reliable technique has been established for simultaneous genotyping of HLA-DPA1 and HLA-DPB1, which may have a broad application in population and disease association studies.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Polimorfismo Genético
/
Datos de Secuencia Molecular
/
Secuencia de Bases
/
Dermatoglifia del ADN
/
Alelos
/
Secuenciación de Nucleótidos de Alto Rendimiento
/
Cadenas alfa de HLA-DP
/
Cadenas beta de HLA-DP
/
Frecuencia de los Genes
/
Genética
Límite:
Humanos
Idioma:
Chino
Revista:
Chinese Journal of Medical Genetics
Año:
2012
Tipo del documento:
Artículo
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