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基于微卫星的聚合酶链反应(PCR)与荧光原位杂交(FISH)检测1p和19q等位基因状态:在患者管理应用中的局限性与优势

Detection of allelic status of 1p and 19q by microsatellite-based PCR versus FISH: limitations and advantages in application to patient management.

作者信息

Jha Prerana, Sarkar Chitra, Pathak Pankaj, Sharma Mehar Chand, Kale Shashank Sharad, Gupta Deepak, Chosdol Kunzang, Suri Vaishali

机构信息

Department of Pathology, All India Institute of Medical Sciences, New Delhi, India.

出版信息

Diagn Mol Pathol. 2011 Mar;20(1):40-7. doi: 10.1097/PDM.0b013e3181e961e9.

DOI:10.1097/PDM.0b013e3181e961e9
PMID:21326038
Abstract

Combined loss of chromosome arms 1p and 19q in oligodendroglial tumors has become a powerful predictor of prognosis and treatment response, and hence clinical testing for their detection is widely used nowadays. Polymerase chain reaction (PCR)-based loss of heterozygosity (LOH) analysis and fluorescence in-situ hybridization (FISH) are the 2 important relatively common clinical molecular diagnostic techniques used for this purpose and they have their unique advantages and limitations. The preference of methodology used depends on local expertise and existing laboratory capabilities. However, there is no consensus on which methodologic approach has a better potential. The objective of the study was to compare the accuracy, reliability, and feasibility of FISH and PCR in detecting the 1p and 19q LOH status. Using the PCR-based method, a LOH analysis was performed on 30 oligodendrogliomas and 10 glioblastomas using fresh-frozen tissue and peripheral blood as control. A FISH assay using paraffin blocks was performed on all the cases. Concordance for 1p and 19q was found in 92.5% (37 of 40) and 82.5% (33 of 40) cases, respectively. The relative advantages and limitations of both the techniques are analyzed and discussed. The main issue pertains to the use of the best technique in large clinical trials whose results are translated to patient care services. Unless the technique used is correct, the results of clinical trials and their correlations may be somewhat questionable.

摘要

少突胶质细胞瘤中1号染色体短臂和19号染色体长臂的联合缺失已成为预后和治疗反应的有力预测指标,因此目前临床检测中广泛应用检测这两种缺失的方法。基于聚合酶链反应(PCR)的杂合性缺失(LOH)分析和荧光原位杂交(FISH)是用于此目的的两种相对常见且重要的临床分子诊断技术,它们各有独特的优势和局限性。所使用方法的偏好取决于当地的专业知识和现有的实验室能力。然而,对于哪种方法更具潜力尚无共识。本研究的目的是比较FISH和PCR检测1p和19q LOH状态的准确性、可靠性和可行性。采用基于PCR的方法,以新鲜冷冻组织和外周血作为对照,对30例少突胶质细胞瘤和10例胶质母细胞瘤进行了LOH分析。对所有病例进行了石蜡切片的FISH检测。1p和19q的一致性分别在92.5%(40例中的37例)和82.5%(40例中的33例)的病例中发现。分析并讨论了这两种技术的相对优势和局限性。主要问题涉及在大型临床试验中使用最佳技术,而这些试验的结果会应用于患者护理服务。除非所使用的技术正确,否则临床试验的结果及其相关性可能会有些可疑。

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