摘要
目的 探讨用化学法（苯酚：氯仿：异戊醇法）从贮存的骨髓涂片中提取微量DNA检测Fms样酪氨酸激酶3内部串联重复（FLT3-ITD）突变情况,分析其对急性髓系白血病（AML）预后的预测价值.方法 采用化学法从58例-20℃贮存1~5年的骨髓涂片（AML 48例和非恶性血液病10例）中提取微量DNA,采用聚合酶链反应（PCR）技术检测患者FLT3-ITD突变情况.结果 48例初治AML患者中6例（12.5%）检测到FLT3-ITD,10例非恶性血液病患者中均未检测到FLT3-ITD.FLT3-ITD阳性AML患者初次诱导完全缓解（CR）率较FLT3-ITD阴性者低[0（0/6）比77.78%（21/27）,P=0.001],总生存时间短（x^2=7.274,P=0.007）.肝、脾大和FLT3突变是影响AML患者初次化疗CR的危险因素（OR=7.2,P=0.12;OR=36.3,P=0.10）.FLT3-ITD是初治AML患者不良预后的危险因素（RR=9.088,P=0.029）.结论 使用化学法提取贮存的AML患者骨髓涂片微量DNA可在临床中应用,是进行回顾性分子生物学研究的重要实验方法.从骨髓涂片中提取微量DNA检测FLT3-ITD可用于预测AML预后.
Objective To investigate the prognostic value of Fms-like tyrosine kinase3, intenal tandem duplication （FLT3-ITD） detection by DNA extracted from stored bone marrow slides in chemical method. Methods Trace DNA was extracted from 58 bone marrow slides which were stored for 1-5 years below 20 ℃, including 48 patients with de novo acute myeloid leukemia （AML） and 10 controls without hematologic malignancies. Polymerase chain reaction （PCR） was used to detect the FLT3-ITD of these bone marrow slides samples. Results There were 6 patients of FLT3-ITD＋ detected in these 48 AML patients （12.5 %, 6/48）. No FLT3-ITD was found in 10 healthy controls. AML patients with FLT3-ITD＋ had low complete time compared with FLT3-ITD-patients （x^2= 7.274, P= 0.007）. Splenohepatomegalia and FLT3 mutation were the risk factors affecting AML patients with CR after the first chemotherapy （OR= 7.2, P=0.12; OR=36.3, P=0.10）. FLT3-ITD was a risk factor of poor prognosis in patients with newly diagnosed AML （RR=9.088, P= 0.029）. Conclusion Extraction of AML bone marrow slides trace DNA by using chemical method can be widely applied in clinic and is a key experimental way to study the molecular biology retrospectively. Furthermore, the detection of FLT3-ITD by trace DNA extracted from stored bone marrow slides can be used to predict the prognosis of AML.
出处
《白血病．淋巴瘤》
CAS
2017年第5期272-275,共4页
Journal of Leukemia & Lymphoma
基金
佛山市卫生和计划生育局医学科研课题（20160126）
