聚合酶链反应检测克隆性免疫球蛋白重链基因重排

目的建立较简便、敏感的Ｂ细胞恶性肿瘤微小残留病（ＭＲＤ）检测方法。方法根据免疫球蛋白重链（ＩｇＨ）基因的保守序列设计引物，应用半巢式聚合酶链反应（ＰＣＲ）基因扩增技术检测克隆性ＩｇＨ基因重排。结果该方法敏感性达１０－３。６２例Ｂ细胞恶性肿瘤患者，５１例检测到克隆性ＩｇＨ基因重排，阳性率为８２％，假阴性率为１８％（１１／６２）。同时检测３５例非Ｂ细胞恶性肿瘤患者和２０例正常人均阴性，无假阳性。检测１４例Ｂ细胞非何杰金淋巴瘤患者形态学检查正常的骨髓标本，ＭＲＤ检出率为７１％（１０／１４）。随访５例处于完全缓解期的Ｂ细胞型急性淋巴细胞白血病患者，６个月内ＭＲＤ均阳性，１２个月后３例转阴性，２例持续阳性，１８个月后持续阳性的２例患者均复发，而转阴性的３例仍处于完全缓解状态。结论半巢式ＰＣＲ检测克隆性ＩｇＨ基因重排，方法简便，敏感性和特异性高，可用于Ｂ细胞恶性肿瘤ＭＲＤ检测

Detection of clonal IgH gene rearrangement by seminested polymerase chain reaction

Objective To establish a method that can detect minimal residual disease (MRD) of B lineage neoplasma. Methods Clonal immunoglobulin heavy chain (IgH) gene rearrangement as a gene marker of B lineage neoplasma, a new method of seminested PCR, was utilized to detect MRD of B lineage neoplasma. Results This method was sensitive to 1 malignant B cell in a background of 10 3 normal cells, with a positive incidence rate of 82%. In addition, although the false negative incidence of this method was 18%, there was no false positive. The application of this method showed that 10 of 14 (71%) B NHL marrow specimens which were normal by morphological method were detected MRD. MRD was also detected in 5 B ALL patients with a complete remission (CR) within 6 months. However in 3 patients with CR, the level of MRD was undetectable for 12, 18 months, while in 2 patients ralapsed in 18 months a trend for MRD persisted at stable levels in 12,18 months. Conclusion Detection of clonal IgH gene rearrangement by seminested PCR provides a simplified, highly specific and sentitive method for detecting MRD of B lineage neoplasma.