筑巢式PCR检测慢性粒细胞白血病bcr-abl融合基因  

Detection of bcr-abl Fusion Gene in Chronic Myelocytic Leukemia Patients by Reverse Transcriptase Polymerse Chain Reation

作　　者：肖敏敏[1] 吴祥林[1] 江继发[2] 

机构地区：[1]安徽省铜陵市人民医院检验科,安徽医科大学在职研究生244000 [2]安徽省铜陵市人民医院血液科,244000

出　　处：《医学研究杂志》2006年第8期19-21,共3页Journal of Medical Research

基　　金：安徽省铜陵市卫生局基金资助项目

摘　　要：目的探讨最适退火温度和核酸扩增（PCR）周期，利用筑巢式PCR检测慢性粒细胞白血病（CML）的bcr-abl融合基因类型，并进行PCR荧光定量（RQ—PCR），研究其与疾病的关系。方法通过改变PCR条件，将退火温度由55℃增加到60℃，反应周期由原来的30周期增加到45周期，检测14例22份标本。同时用荧光定量试剂检测标本。结果退火温度为58℃，45周期可测出10^3 copies／μl定量标准品的PCR产物。22份标本6cr-abl融合基因巢式PCR结果均阳性。其中b2a2型9份，b3a2型13份。定量检测18例阳性，量值在10^2-10^6copies／μg，两例急变患者急性期和慢性期有明显差异。结论筑巢式PCR是一种敏感而准确的检测方法，对bcr—abl进行定量有助于反映白血病细胞负荷、评价疗效及判断疾病预后。Objective To increase the sensitivity of residual leukemic cells detectin in chronic myelocytic leukemia（CML） patients with RT - PCR, the optional annealing temperature and PCR cycles were studied to confirm bcr - abl fused gene types, and bcr - abl mRNA transcripts were monitored by RQ -PCR to study the relation with CML at different phases. Methods Through changing the PCR conditions, the annealing temperature was measured from 55℃ to 60℃, and the number of reaction cycles was increased from 30 to 45. All 22 samples were examined, and bcr - abl mRNA transcripts were quantified by RQ - PCR kit. Results Bcr - abl fused gene types were found in 22 samples,of all 9 cases were b2a2 type, 13 cases were b3a2. When the annealing temperature was set for 58℃ and the number of reation cycles was set for 45,103 copies/ul standard samples was detected. 18 samples were positive tested by RQ -PCR kit, and the value was between 102 to 106 copies/g. There were significant differce between the results of chronic phase samples and those of accelerated phase. Conclusions The RT - PCR is a reliable, sensitive and reproducible method of monitoring CML patients. The real - time RT - PCR is useful in evaluating leukemic burden, assessing response to treatment and predicating the prognosis of the disease.

关 键 词：慢性粒细胞白血病 聚合酶链反应 bcr—abl融合基因 

分 类 号：R733.72[医药卫生—肿瘤] R512.62[医药卫生—临床医学]