白血病患者循环DNA定量和完整性分析及其临床意义

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目的分析白血病患者血浆DNA的含量及完整性,并探讨其临床意义。方法以2008年6月-2009年3月收治的60例白血病患者作为实验组,同期在门诊行体检的健康正常人20例、血液系统良性病变者20例、淋巴瘤患者10例、实体肿瘤患者20例(食管癌6例、原发性肝癌4例、鼻咽癌5例、乳腺癌5例)作为对照组(n=70)。测定两组患者的血浆DNA含量,比较两组间、不同FAB分型的急性髓细胞白血病患者间、实验组不同年龄段及不同病程患者间血浆DNA水平的差异。采用聚合酶链反应扩增β-actin大片段和小片段产物并行琼脂糖凝胶电泳观察,比较实验组患者与对照组中正常人血浆DNA的差异。结果实验组患者血浆DNA浓度(413.2±118.4μg/L)约为对照组中正常人(19.6±7.9μg/L)的21倍(P<0.05),且高于对照组的其他疾病患者(P<0.05)。实验组血浆DNA浓度与年龄及疾病病程有关,不同FAB型别的急性髓细胞白血病患者血浆DNA含量无显著差异。实验组血浆DNA电泳呈现大小不一的片段,而且其血浆DNA中可扩增出β-actin的大、小片段产物,完整性较高。正常人仅可见少量小片段DNA。结论白血病患者血浆DNA含量和完整性明显增高,对白血病的诊断及预后判断具有重要价值。 Objective To analyze the content and integrity of plasma DNA in patients with leukemia and to explore its clinical significance. Methods Sixty leukemia patients admitted from June 2008 to March 2009 were selected as the experimental group. In the same period, 20 healthy subjects were examined in the outpatient setting, 20 with benign disease in the blood system, 10 with lymphomas, 20 cases (esophageal cancer in 6 cases, 4 cases of primary liver cancer, 5 cases of nasopharyngeal carcinoma, 5 cases of breast cancer) as a control group (n = 70). The plasma DNA levels were measured in two groups. The differences of plasma DNA levels between two groups and among patients with acute myeloid leukemia with different FAB typing and between different age groups and different durations of the experimental group were compared. Polymerase chain reaction amplification of β-actin large fragments and small fragments of products were observed by agarose gel electrophoresis, the experimental group patients and control group, the difference between normal plasma DNA. Results The plasma DNA concentration in the experimental group (413.2 ± 118.4μg / L) was about 21 times higher than that in the control group (19.6 ± 7.9μg / L) (P <0.05), and higher than that in the control group <0.05). Plasma DNA concentration in the experimental group was related to age and disease duration. There was no significant difference in plasma DNA levels between different FAB types of acute myeloid leukemia patients. Plasma DNA electrophoresis in the experimental group showed fragments of different sizes, and large and small fragment products of β-actin were amplified in the plasma DNA, and the integrity was high. Normal people can only see a small amount of small pieces of DNA. Conclusion The plasma DNA content and integrity of patients with leukemia are significantly increased, which is of great value in the diagnosis and prognosis of leukemia.
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