Qi Zong-Li, Zhang Bao, Han Xi-Qun, Zhu Mei-Gang, Zhao Tong
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2008 Nov;28(11):1964-7.
To analyze and optimize the gene rearrangement primers of different frame regions (FR) of immunoglobulin heavy chain (IgH) genes by bioinformatic methods and explore the application of these primers in the detection of paraffin-embedded lymphoma tissues.
Three pairs of primers from IgH FR1, FR2 and FR3 regions (P1c, P2A and P31, respectively) were selected as the B cell gene rearrangement primers after comparison of the gene fragments in 44 IgH variable and 6 joining regions. Using one pair of T cell receptor (TCR) gamma primer as the T cell gene rearrangement primer, 101 histopathologically confirmed lymphoproliferative samples including 80 B cell lymphomas, 14 T cell lymphomas, and 7 reactive proliferative lymph nodes were examined by PCR for gene arrangement. The DNAs from DG75 and Jurkat cell lines were used as the positive controls for B and T cell lymphoma, respectively, with those from reactive proliferative lymph nodes as the negative control.
The positivity rates of IgH primers (P1c, P2A and P31) in the 80 B cell lymphomas were 37.5% (30/80), 52.5% (42/80) and 70.0% (56/80), respectively, and only one of the 14 T cell lymphoma cases was positive for the primers, suggesting significant differences in the detection rates of B cell lymphomas by the 3 primers. The detection rate was increased to 83.9% by combining the results by P31 and P2A primers. No positivity was found in the proliferative reaction tissues.
Primers from IgH FR3 region genes are more sensitive than that from the FR1 and FR2 regions in the detection of gene rearrangement in paraffin-embedded lymphoma tissues. The detection rates can be increased by combining the results with the primers for IgH FR3 with that of FR2.
运用生物信息学方法分析并优化免疫球蛋白重链（IgH）基因不同框架区（FR）的基因重排引物，探讨这些引物在石蜡包埋淋巴瘤组织检测中的应用。
在比较44个IgH可变区和6个连接区的基因片段后，选择来自IgH FR1、FR2和FR3区的三对引物（分别为P1c、P2A和P31）作为B细胞基因重排引物。使用一对T细胞受体（TCR）γ引物作为T细胞基因重排引物，通过聚合酶链反应（PCR）检测101例经组织病理学确诊的淋巴增殖性样本，其中包括80例B细胞淋巴瘤、14例T细胞淋巴瘤和7例反应性增生淋巴结的基因重排情况。分别以DG75和Jurkat细胞系的DNA作为B细胞和T细胞淋巴瘤的阳性对照，以反应性增生淋巴结的DNA作为阴性对照。
80例B细胞淋巴瘤中，IgH引物（P1c、P2A和P31）的阳性率分别为37.5%（30/80）、52.5%（42/80）和70.0%（56/80），14例T细胞淋巴瘤病例中只有1例对这些引物呈阳性，表明这3种引物对B细胞淋巴瘤的检测率存在显著差异。将P31和P2A引物的结果相结合，检测率提高到了83.9%。在增生反应组织中未发现阳性结果。
在石蜡包埋淋巴瘤组织的基因重排检测中，IgH FR3区基因的引物比FR1和FR2区的引物更敏感。将IgH FR3区引物与FR2区引物的结果相结合可提高检测率。
