Development of a high-throughput sequence-based typing assay for human leukocyte antigen loci / 中华医学遗传学杂志
Zhonghua Yi Xue Yi Chuan Xue Za Zhi
; (6): 323-327, 2012.
Article
de Zh
| WPRIM
| ID: wpr-295484
Bibliothèque responsable:
WPRO
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*02:02 (0.589) > DPA1*01:03 (0.284) > DPA1*02:01 (0.096) > DPA1*04:01 (0.031) were identified. In addition, 14 HLA-DPB1 alleles, including 4 common alleles (with a frequency of more than 5%, namely DPB1*05:01, DPB1*02:01, DPB1*04:01 and DPB1*02:02), 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>
Texte intégral:
1
Indice:
WPRIM
Sujet Principal:
Polymorphisme génétique
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Données de séquences moléculaires
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Séquence nucléotidique
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Profilage d'ADN
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Allèles
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Séquençage nucléotidique à haut débit
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Chaines alpha des antigènes HLA-DP
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Chaines bêta des antigènes HLA-DP
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Fréquence d'allèle
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Génétique
Limites du sujet:
Humans
langue:
Zh
Texte intégral:
Zhonghua Yi Xue Yi Chuan Xue Za Zhi
Année:
2012
Type:
Article