Quantitative real-time PCR method for detection of B-lymphocyte monoclonality by comparison of κ and λ immunoglobulin light chain expression
Quantitative real-time PCR method for detection of B-lymphocyte monoclonality by comparison of κ and λ immunoglobulin light chain expression
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DOI：
10.1373/49.1.51
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发表时间：
2003-01-01
影响因子：
9.3
通讯作者：
Kubista, M
中科院分区：
文献类型：
--
作者：
Ståhlberg, A;Åman, P;Kubista, M
背景：IgL κ：IgL λ比值异常长期以来一直被用作非霍奇金B细胞淋巴瘤的临床标准。作为对淋巴瘤的定量实时PCR为基础的多标记物诊断分析的第一步，我们已经开发了一种方法，用于确定IgLkappa：IgLlambda比率在临床samples.Methods：点亮探针为基础的实时PCR被用来定量IgLkappa和IgLlambda cDNA从32个临床样本。样本也进行了研究，通过常规免疫组化分析和流式细胞仪。结果：32例疑似非霍奇金淋巴瘤的样本分析，28个被正确分配从实时PCR测量假设不变的PCR效率的生物样品。4份样本为假阴性。一个是T细胞淋巴瘤，一个是弥漫性大B细胞淋巴瘤，一个是重新分析，并发现淋巴瘤阳性的原位校准，其中考虑到样本特异性PCR抑制。12个样品是细针抽吸，这些都是正确的assigned.Conclusions：这项工作是第一步，通过定量点亮探针为基础的实时PCR分析临床样本。定量实时PCR似乎适合通过测量细针抽吸物中肿瘤标志物的表达来高通量检测癌症。(C)2003年美国临床化学协会。
Background: An abnormal IgLkappa:IgLlambda ratio has long been used as a clinical criterion for non-Hodgkin B-cell lymphomas. As a first step toward a quantitative real-time PCR-based multimarker diagnostic analysis of lymphomas, we have developed a method for determination of IgLkappa:IgLlambda ratio in clinical samples.Methods: Light-up probe-based real-time PCR was used to quantify IgLkappa and IgLlambda cDNA from 32 clinical samples. The samples were also investigated by routine immunohistochemical analysis and flow cytometry.Results: Of 32 suspected non-Hodgkin lymphoma samples analyzed, 28 were correctly assigned from real-time PCR measurements assuming invariant PCR efficiencies in the biological samples. Four samples were false negatives. One was a T-cell lymphoma, one was a diffuse large B-cell lymphoma, and one was reanalyzed and found lymphoma-positive by in situ calibration, which takes into account sample-specific PCR inhibition. Twelve of the samples were fine-needle aspirates, and these were all correctly assigned.Conclusions: This work is a first step toward analyzing clinical samples by quantitative light-up probe-based real-time PCR. Quantitative real-time PCR appears suitable for high-throughput testing of cancers by measuring expression of tumor markers in fine-needle aspirates. (C) 2003 American Association for Clinical Chemistry.