Microdissection and gene rearrangement analysis of paraffin-embedded specimens of orbital malignant lymphoma.
Microdissection and gene rearrangement analysis of paraffin-embedded specimens of orbital malignant lymphoma.
复制标题
眼眶恶性淋巴瘤石蜡包埋标本的显微切割和基因重排分析。
DOI：
10.1007/s10384-003-0038-7
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发表时间：
2004
影响因子：
2.4
通讯作者：
Mochizuki,Manabu
中科院分区：
文献类型：
--
作者：
Miyanaga,Masaru;Kiyosawa,Motohiro;Takase,Hiroshi;Eishi,Yoshinobu;Shen,DeFen;Chan,Chi-Chao;Mochizuki,Manabu
目的应用显微解剖和聚合酶链反应（PCR）技术对石蜡包埋的眼眶标本进行基因重排分析，探讨恶性淋巴瘤的诊断价值。方法对4例经组织病理学诊断为眼眶恶性淋巴瘤的患者进行标本检查。从石蜡包埋标本上显微解剖恶性细胞。提取细胞DNA， PCR扩增免疫球蛋白重链（immunoglobulin heavy chain, IgH）基因。利用第三框架（FR3A）、第二框架（FR2A）和互补决定区3 （CDR3）的引物检测基因重排。还检测了b细胞淋巴瘤/白血病-2 （bcl-2）基因的易位。结果4例石蜡包埋标本中，3例载玻片上可见恶性细胞。这三个病例的标本显示FR3A、FR2A和CDR3的重排。在1例中也检测到abcl -2相关易位。结论石蜡包埋眼窝标本的基因重排分析可用于恶性淋巴瘤的诊断。该方法的优点是由于检测灵敏度高，只需要一个小样本。中华眼科杂志2004；48:123-127©日本眼科学会2004
PurposeTo determine whether a definite diagnosis of malignant lymphoma can be made from paraffin-embedded archived orbital specimens by gene rearrangement analysis using microdissection and polymerase chain reaction (PCR).MethodsSpecimens from four patients with histopathologically diagnosed orbital malignant lymphoma were examined. The malignant cells were microdissected off the paraffin-embedded specimens. DNA was extracted from the cells, and the immunoglobulin heavy chain (IgH) gene was amplified by PCR. Gene rearrangements were detected by using primers for the third framework (FR3A), the second framework (FR2A), and the complementary determining region 3 (CDR3). Translocation of the B-cell lymphoma/leukemia-2 (bcl-2) gene was also examined.ResultsMalignant cells were present on the slides of the paraffin-embedded specimens of three of four cases. The specimens from these three cases showedIgHrearrangements for FR3A, FR2A, and CDR3. Abcl-2-associated translocation was also detected in one case.ConclusionsGene rearrangement analysis is applicable to paraffin-embedded archived orbital specimens to confirm a diagnosis of malignant lymphoma. The advantage of this method is that only a small specimen is needed because the detection sensitivity is high.Jpn J Ophthalmol2004;48:123–127 © Japanese Ophthalmological Society 2004