Nikiforova Marina N, Hsi Eric D, Braziel Rita M, Gulley Margaret L, Leonard Debra G B, Nowak Jan A, Tubbs Raymond R, Vance Gail H, Van Deerlin Vivianna M
Department of Pathology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
Arch Pathol Lab Med. 2007 Feb;131(2):185-9. doi: 10.5858/2007-131-185-DOCIGR.
The diagnosis of B-cell lymphoid malignancy can frequently be substantiated by detecting clonal immunoglobulin heavy chain (IGH) gene rearrangements, which is typically done by polymerase chain reaction (PCR) amplification and/or Southern blot analysis.
To characterize current laboratory practice for the assessment of IGH rearrangements and to identify opportunities for improvement.
The data from the Molecular Oncology Proficiency Survey distributed to participating laboratories by the Molecular Pathology Committee of the College of American Pathologists from 1998 through 2003 were analyzed.
Thirty-nine proficiency survey specimens (29 positive and 10 negative for clonal IGH rearrangements) were distributed. For Southern blot analysis, 944 results were reported, with a successful response rate of 95%. For PCR detection, 2349 results were reported, with a successful response rate of 72%. A higher rate of successful responses by PCR was achieved using framework 3 primers in combination with other frameworks (82%) compared with framework 3 primers only (76%) and when fresh/frozen (72%) compared with paraffin-embedded (65%) tissues were analyzed.
The performance of the participating laboratories was very good, by both Southern blot and PCR analysis. As expected, Southern blot analysis consistently detects a higher proportion of IGH rearrangements than PCR analysis. Further improvement and standardization of the IGH PCR assay is important if it is to replace Southern blot analysis as the standard method. Participation in this survey is a valuable tool for assessing laboratory performance and it directs our attention to areas where we may improve laboratory practice.
B细胞淋巴瘤的诊断通常可通过检测克隆性免疫球蛋白重链（IGH）基因重排来证实，这通常通过聚合酶链反应（PCR）扩增和/或Southern印迹分析来完成。
描述当前评估IGH重排的实验室操作，并确定改进的机会。
分析了美国病理学家学会分子病理学委员会从1998年至2003年分发给参与实验室的分子肿瘤学能力验证调查的数据。
共分发了39份能力验证标本（29份克隆性IGH重排阳性，10份阴性）。对于Southern印迹分析，报告了944个结果，成功应答率为95%。对于PCR检测，报告了2349个结果，成功应答率为72%。与仅使用框架3引物（76%）相比，使用框架3引物与其他框架组合（82%）时，PCR的成功应答率更高；与分析石蜡包埋组织（65%）相比，分析新鲜/冷冻组织（72%）时，PCR的成功应答率更高。
通过Southern印迹和PCR分析，参与实验室的表现都非常好。正如预期的那样，Southern印迹分析始终比PCR分析检测到更高比例的IGH重排。如果IGH PCR检测要取代Southern印迹分析作为标准方法，进一步改进和标准化该检测方法很重要。参与这项调查是评估实验室表现的宝贵工具，它使我们关注到可以改进实验室操作的领域。
