CAJ | 학술논문

目的改进实时定量RT-PCR的检测计算方法,提高定量RT-PCR在急性早幼粒细胞白血病(APL)微量残留病变监测的临床应用价值.方法采用实时定量RT-PCR和常规定性RT-PCR检测31例APL患者在治疗前后融合基因PML-RARα的表达水平,对两种计算方法进行比较:常规按照患者治疗前后自身比较计算(常规法)和治疗前标准化基线水平计算(标化法).结果31例患者采用两种定量计算方法结果近似,标化法计算结果示患者在治疗缓解、巩固治疗和维持治疗期间融合基因PML-RARα转录本对数下降值分别为(2.0±1.9)、(4.9±1.4)和(5.7±0.1),常规法计算结果为(1.9±1.9)、(4.8±1.3)和(5.7±0.4).在维持治疗阶段标化法定量RT-PCR的结果显示患者个体差异更小.按照标化法计算结果,次要分子生物反应(对数值≥3)和主要分子生物反应(对数值≥5)标准仍然适用.结论采用标化法计算实时定量RT-PCR结果能较好的对APL患者的微量残留病变进行监测,一定程度上降低了患者个体差异对检测结果的影响,同时适用于治疗前标本缺如的患者.
목적 개진실시정량RT-PCR적검측계산방법,제고정량RT-PCR재급성조유립세포백혈병(APL)미량잔류병변감측적림상응용개치.방법 채용실시정량RT-PCR화상규정성RT-PCR검측31례APL환자재치료전후융합기인PML-RARα적표체수평,대량충계산방법진행비교:상규안조환자치료전후자신비교계산(상규법)화치료전표준화기선수평계산(표화법).결과31례환자채용량충정량계산방법결과근사,표화법계산결과시환자재치료완해、공고치료화유지치료기간융합기인PML-RARα전록본대수하강치분별위(2.0±1.9)、(4.9±1.4)화(5.7±0.1),상규법계산결과위(1.9±1.9)、(4.8±1.3)화(5.7±0.4).재유지치료계단표화법정량RT-PCR적결과현시환자개체차이경소.안조표화법계산결과,차요분자생물반응(대수치≥3)화주요분자생물반응(대수치≥5)표준잉연괄용.결론 채용표화법계산실시정량RT-PCR결과능교호적대APL환자적미량잔류병변진행감측,일정정도상강저료환자개체차이대검측결과적영향,동시괄용우치료전표본결여적환자.
Objective To optimize the calculation of quantitative real time RT-PCR (Q-RT-PCR) of PML-RARα in patients with acute promyelocytic leukemia (APL) for molecular monitoring of minimal residual disease(MRD). Methods By using both regular reverse transcription polymerase chain reaction (RT-PCR) and Q-RT-PCR, the expression levels of PML-RARα transcripts were measured before and after treatment. The conventional Q-RT-PCR calculation was directly compared the post-treatment transcript level with the respective pre-treatment one (DoseN) in the individual patient while the standardized calculation was based on the calculation of standardized pre-treatment DoseN of all patients. Results In 181 samples from 31 patients, the results of log-reduction of PML-RARα after induction, at the end of consolidation and during maintenance by conventional method were ( 1.9±1.9), (4.8±1.3 ) and (5.7±0.4), respectively, while by standardized method were (2.0±1.9), (4.9±1.4) and (5.7±0.1 ), respectively. Of notice, the result was with significant less variation of the latter methods during maintenance therapy. Moreover, with defined criteria of molecular response (3.0-4.9 log-reduction as minor and≥5.0 log-reduction as major molecular response), the standardized method was validated in clinical settings. Conclusion The standardized method is superior to the conventional method for calculation of Q-RT-PCR results. The new method can reduce the individual variation in monitoring the MRD and is feasible even for patients with unavailable pretreatment samples.