Objective: To improve the positivity rate and accuracy of MYD88 mutation detection in patients with Waldenström macroglobulinemia (WM) . Methods: MYD88 mutation status was retrospectively evaluated in 66 patients diagnosed with WM in Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from June 2017 to June 2021. The positivity rate and accuracy of the different methods and specimens for MYD88 mutation detection were analyzed. Results: MYD88 mutations were detected in 51 of 66 patients with WM, with an overall positivity rate of 77%. The positivity rate of the next-generation sequencing (NGS) or allele-specific polymerase chain reaction (AS-PCR) was significantly higher than that of the first-generation Sanger sequencing (84% vs 71% vs 46%, P<0.05) . For the different specimens, the positivity rate for the lymph nodes or bone marrow was significantly higher than that of peripheral blood (79% vs 84% vs 52%, P<0.05) . The positivity rate of the MYD88 mutation in the lymph nodes, bone marrow, and peripheral blood determined by NGS was 86%, 90%, and 67%, respectively. The positivity rate in the lymph nodes, bone marrow, and peripheral blood detected by AS-PCR was 78%, 81%, and 53%, respectively. Thirty-nine patients with WM underwent ≥ 2 MYD88 mutation detections. The final MYD88 mutational status for each patient was used as the standard to determine the accuracy of the different methods and in different specimens. The accuracy of MYD88 mutation detection in the lymph nodes (n=18) and bone marrow (n=13) by NGS was significantly higher than that in the peripheral blood (n=4) (100% vs 100% vs 75%, P<0.05) . There was no statistically significant difference in the accuracy of MYD88 mutation detection by AS-PCR in the lymph nodes (n=15) , bone marrow (n=11) , or peripheral blood (n=16) (93% vs 91% vs 88%, P>0.05) . Conclusions: In the detection of the MYD88 mutation in patients diagnosed with WM, NGS or AS-PCR is more sensitive than Sanger sequencing. Lymph nodes and bone marrow specimens are better than peripheral blood specimens.
目的： 探索如何提高华氏巨球蛋白血症（WM）患者MYD88突变检测的阳性率和准确率。 方法： 回顾性分析2017年6月至2021年6月上海交通大学医学院附属瑞金医院66例初诊WM患者MYD88突变检测结果，分析不同方法和标本检测MYD88突变的阳性率和准确率。 结果： 66例WM患者中51例MYD88突变阳性，整体阳性率77%。根据检测方法分类：二代测序（NGS）和等位基因特异性PCR（AS-PCR）检测突变阳性率显著高于一代测序Sanger法（84%对71%对46%，P<0.05）。根据标本取材部位分类：淋巴结和骨髓标本检测突变阳性率显著高于外周血标本（79%对84%对52%，P<0.05）。NGS检测淋巴结、骨髓和外周血MYD88突变阳性率分别为86%、90%和67%。AS-PCR检测淋巴结、骨髓和外周血MYD88突变阳性率分别为78%、81%和53%。39例WM患者进行≥2次MYD88突变检测，以每例患者最终突变检测结果作为标准，判断不同方法和标本的准确率。淋巴结（18例）和骨髓（13例）NGS检测准确率显著高于外周血（4例）标本（100%对100%对75%，P<0.05）。淋巴结（15例）、骨髓（11例）和外周血（16例）AS-PCR检测准确率的差异无统计学意义（93%对91%对88%，P>0.05）。 结论： 在检测WM患者MYD88突变情况方面，NGS或AS-PCR法优于Sanger法，淋巴结和骨髓标本优于外周血标本。.
Keywords: Myeloid differentiation factor 88; Next-generation sequencing; Polymerase chain reaction; Waldenstrom Macroglobulinemia.