文章：

多参数流式细胞术与增强型ASO RQ-PCR在多发性骨髓瘤微小残留病检测中的比较分析

Comparative analysis of minimal residual disease detection by multiparameter flow cytometry and enhanced ASO RQ-PCR in multiple myeloma

原文发布日期：2014-10-10

DOI: 10.1038/bcj.2014.69

类型: Original Article

开放获取: 是

英文摘要：

Multiparameter flow cytometry (MFC) and allele-specific oligonucleotide real-time quantitative PCR (ASO RQ-PCR) are the two most sensitive methods to detect minimal residual disease (MRD) in multiple myeloma (MM). We compared these methods in 129 paired post-therapy samples from 22 unselected, consecutive MM patients in complete/near complete remission. Appropriate immunophenotypic and ASO RQ-PCR-MRD targets could be detected and MRD analyses constructed for all patients. The high PCR coverage could be achieved by gradual widening of the primer sets used for clonality detection. In addition, for 13 (55%) of the patients, reverse orientation of the ASO primer and individual design of the TaqMan probe improved the sensitivity and specificity of ASO RQ-PCR analysis. A significant nonlinear correlation prevailed between MFC-MRD and PCR-MRD when both were positive. Discordance between the methods was found in 32 (35%) paired samples, which were negative by MFC-MRD, but positive by ASO RQ-PCR. The findings suggest that with the described technique, ASO RQ-PCR can be constructed for all patients with MM. ASO RQ-PCR is slightly more sensitive in MRD detection than 6−10-color flow cytometry. Owing to technical demands ASO RQ-PCR could be reserved for patients in immunophenotypic remission, especially in efficacy comparisons between different drugs and treatment modalities.

摘要翻译： 

多参数流式细胞术（MFC）和等位基因特异性寡核苷酸实时定量PCR（ASO RQ-PCR）是多发性骨髓瘤（MM）中检测微小残留病（MRD）的两种最灵敏方法。我们在22例未经筛选、连续达到完全/接近完全缓解的MM患者中，对129份治疗后配对样本进行了两种方法的比较。所有患者均能检测到合适的免疫表型和ASO RQ-PCR MRD靶标，并完成MRD分析。通过逐步扩大用于克隆性检测的引物组范围，可实现较高的PCR覆盖率。此外，对13例（55%）患者采用反向ASO引物和个性化设计的TaqMan探针，提升了ASO RQ-PCR分析的灵敏度和特异性。当两种检测均为阳性时，MFC-MRD与PCR-MRD之间存在显著非线性相关性。32份（35%）配对样本存在方法学差异：这些样本经MFC-MRD检测为阴性，但ASO RQ-PCR检测为阳性。研究结果表明采用所述技术可为所有MM患者构建ASO RQ-PCR检测体系。在MRD检测方面，ASO RQ-PCR比6-10色流式细胞术灵敏度略高。鉴于技术要求，ASO RQ-PCR可适用于免疫表型缓解期患者，特别是在不同药物与治疗模式的疗效比较中。

原文链接：

Comparative analysis of minimal residual disease detection by multiparameter flow cytometry and enhanced ASO RQ-PCR in multiple myeloma