中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (33): 6194-6198.doi: 10.3969/j.issn.2095-4344.2012.33.023

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

中国汉族人群鼻咽癌患者HLA-DQB1等位基因多态性分析

卢 亮1，吕 震2   

  1. 1深圳市血液中心，广东省深圳市 518035；
    2蚌埠医学院，安徽省蚌埠市 233000
  • 收稿日期:2012-01-17 修回日期:2012-05-08 出版日期:2012-08-12 发布日期:2012-08-12
  • 通讯作者: 卢亮★，男，广东省广州市人，汉族，1989年中山大学医疗系毕业，硕士，副主任技师，主要从事输血医学检验方面研究。 luliang189@163.com
  • 作者简介:卢亮★，男，广东省广州市人，汉族，1989年中山大学医疗系毕业，硕士，副主任技师，主要从事输血医学检验方面研究。 luliang189@163.com

Polymorphism of HLA-DQB1 alleles in nasopharyngeal carcinoma patients

Lu Liang1, Lü Zhen2   

  1. 1Shenzhen Blood Center Institute, Shenzhen HLA Laboratory of China Marrow Donor Program, Shenzhen 518035, Guangdong Province, China;
    2 Bengbu Medical College, Bengbu 233000, Anhui Province, China
  • Received:2012-01-17 Revised:2012-05-08 Online:2012-08-12 Published:2012-08-12
  • Contact: Lu Liang, Shenzhen Blood Center Institute, Shenzhen HLA Laboratory of China Marrow Donor Program, Shenzhen 518035, Guangdong Province, China luliang189@163.com
  • About author:Lu Liang★, Master, Associate chief technician, Shenzhen Blood Center Institute, Shenzhen HLA Laboratory of China Marrow Donor Program, Shenzhen 518035, Guangdong Province, China

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摘要：

背景：目前开展HLA-DQB1基因与鼻咽癌疾病相关性研究一方面受昂贵的进口试剂所制约，另一方面单纯做HLA-DQB1第2外测序分型，模棱两可，结果较少。
目的：采用HLA-DQB1基因第2至第3外显子双向测序分型检测技术方法。
方法：纳入286份中国汉族健康人群血液样本，46例鼻咽癌患者样本，使用Gentra DNA提取试剂制备基因组DNA，取100份健康人群样本作为平行对照组进行基因分型，选择及自行设计HLA-DQB1基因第2至3外显子扩增引物及测序引物，探索最适合PCR扩增及测序反应条件。采用商品化HLA-DQB1基因单向测序分型试剂盒作对照。
背景：目前开展HLA-DQB1基因与鼻咽癌疾病相关性研究一方面受昂贵的进口试剂所制约，另一方面单纯做HLA-DQB1第2外测序分型，模棱两可，结果较少。
目的：采用HLA-DQB1基因第2至第3外显子双向测序分型检测技术方法。
方法：纳入286份中国汉族健康人群血液样本，46例鼻咽癌患者样本，使用Gentra DNA提取试剂制备基因组DNA，取100份健康人群样本作为平行对照组进行基因分型，选择及自行设计HLA-DQB1基因第2至3外显子扩增引物及测序引物，探索最适合PCR扩增及测序反应条件。采用商品化HLA-DQB1基因单向测序分型试剂盒作对照。
结果与结论：100份平行对照组样本与HLA-SBT测序分型结果一致。286例健康个体中共检出13种等位基因，27例样本的等位基因为纯合子。鼻咽癌人群共检出DQB1等位基因11种，5例样本的等位基因为纯合子。鼻咽癌患者DQB1*02等位基因频率高于对照组，但所有等位基因频率比较，差异无显著性意义(χ2 < 3.84，P > 0.05)。未发现HLA-DQB1基因与鼻咽癌有相关性。说明采用的中国汉族人群HLA-DQB1基因测序分型方法具有操作方便、结果稳定和低成本消耗等优势。100份平行对照组样本与HLA-SBT测序分型结果一致。286例健康个体中共检出13种等位基因，27例样本的等位基因为纯合子。鼻咽癌人群共检出DQB1等位基因11种，5例样本的等位基因为纯合子。鼻咽癌患者DQB1*02等位基因频率高于对照组，但所有等位基因频率比较，差异无显著性意义(χ2 < 3.84，P > 0.05)。未发现HLA-DQB1基因与鼻咽癌有相关性。说明采用的中国汉族人群HLA-DQB1基因测序分型方法具有操作方便、结果稳定和低成本消耗等优势。

关键词: 人类白细胞抗原, 基因型, 中国汉族, 基因测序, 鼻咽癌

Abstract:

BACKGROUND: Correlation studies between HLA-DQB1 gene and nasopharyngeal carcinoma are constrained by expensive imported reagents; on the other hand, the genotyping of HLA-DQB1 at exon 2 is ambiguous, and the results are less.
OBJECTIVE: To apply the direct sequencing genotyping method of HLA-DQB1 gene at exons 2 and 3.
METHODS: 286 samples from voluntary healthy donors and 46 samples from nasopharyngeal carcinoma patients were included. Gentra DNA extraction reagent was applied to prepare genomic DNA. 100 samples from healthy donors served as parallel control group for genotyping. We explored the most suitable conditions for PCR amplification and sequencing reaction of HLA-DQB1 allele exons 2 and 3. Post-process HLA-DQB1 gene sequencing parting kit was used for one-way parallel controls.
RESULTS AND CONCLUSION: The results of 100 samples as the parallel control are consistent with those of HLA-SBT genotyping. For 286 healthy samples, 13 HLA-DQB1 alleles were detected and 27 HLA-DQB1 alleles were homozygous in unrelated healthy individuals. Eleventh HLA-DQB1 alleles were detected, and five HLA-DQB1 alleles were homozygous in nasopharyngeal carcinoma patients. The frequency of HLA-DQB1*02:01 in nasopharyngeal carcinoma C patients was higher than that in unrelated healthy individuals; however, the frequency distribution of HLA-DQB1 alleles had not significant differences between the two groups (x2 < 3.84, P > 0.05). No significant association between the HLA-DQB1 and nasopharyngeal carcinoma was confirmed. These findings indicate that the method for HLA-DQB1 genotype from the Chinese Han population is easy to operate with stable results and low cost.

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