巢式PCR扩增TCR Vδ_2-Dδ_3重排片段检测急淋脑脊液微量白血病细胞

Determination of minimal leukenic cells in cerebrospinal fluid by nested PCR amplifying TCR V6j-V6, rearranged fragment In acute childhood lymphoblastic leukemia

为探索急性淋巴细胞白血病(ALL)患儿脑脊液(CSF)中微量白血病细胞的检测方法及其意义,本文采用巢式聚合酶链反应(PCR)扩增T细胞受体的Vδ_2-Dδ_3重排片段方法,对21侧小儿ALL进行检测。结果:(1)改良一步煮沸法抽提CSF的DNA效果良好,巢式PCR方法敏感、可靠:(2)13例初治病例中,CSF的微量白血病细胞阳性6例,经常规三联鞘注及大剂量氨甲喋啶(HDMTX)治疗后4例转为阴性并持续缓解,另2例阳性者随之发生中枢神经系统白血病(CNSL);(3)8例缓解治疗期ALL中7例追踪结果,3例经鞘注和HD MTX治疗后CSF和BM的微量白血病细胞同为阳性者出现骨髓复发。结果提示对CSF的微量白血病细胞动态监测对指导治疗、预示CNSL的发生有重要意义。

In order to explore the availability and its clinical significance for detecting the minimal leukemic cells of CSF in patients with acute lymphoblastic leukemia (ALL), 21 ALL cases were determined with nested PCR amplifying TCR Vδ2-Vδ3 rearranged fragment. The results showed that:a) there was a fine effect to extract DNA in CSF by improving one-step boiling procedure and the nested PCR amplifying TCR Vδ2-Vδ3 rearranged fragment was sensitive and reliable, b) The positive minimal leukemic cells in CSF were found in 6 cases of 13 untreated patients. Four cases of the six positive patients turned to be the negative results in CSF with persistent CR after treatment with routine intrathecal injection of three-drug combination and the use of high dose of MTX. CNSL occurred in the rest of two positive cases who were with unchanged positive results in CSF. c) Of 8 patients with ALL in the CR period, 7 cases were followed up clinically. The results showed that three cases with positive minimal leukemic cells in both CSF and bone marrow relapse in bone marrow. Suggesting that dynamic monitoring minimal leukemic cells in CSF could be valuable in guiding the management of ALL and predicting the involevement of CNSL.