EB病毒与急性白血病的相关性研究

发布时间：2018-03-20 21:34

本文选题：EB病毒　切入点：实时荧光定量聚合酶链式反应　出处：《青岛大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的检测不同类型白血病(急性淋巴细胞白血病、急性髓细胞白血病)患者EB病毒(EBV)的感染情况,以探讨EBV与白血病的发生、发展及细胞遗传学改变的相关性,为揭示EBV感染与某些类型白血病的发生密切相关提供可靠依据。方法收集185例白血病患者作为病例组,其中110例为急性淋巴细胞白血病(ALL),75例为急性髓细胞白血病(AML),同时收集37例正常人(“正常”指没有任何类型的血液系统疾病和其他癌症且骨髓象大致正常)作为对照组,采用常规苯酚-氯仿-异戊醇方法提取骨髓细胞DNA,采用实时荧光定量聚合酶链反应(FQ-PCR)检测所有标本骨髓细胞中EBV-DNA;采用骨髓液短期培养法制备染色体标本并采用常规R显带技术分析核型;运用单克隆抗体技术对ALL进行免疫分型,进一步分析T-ALL和B-ALL与EB病毒感染的相关性,并对部分患者进行了临床随访,以进一步探讨EB病毒感染对白血病患者的复发及死亡的影响。两独立样本定量资料的检验应用Mann-Whitney U非参数检验方法;各组间EBV感染率的比较采用χ2检验;Log-rank检验用于比较EBV感染与ALL患者之间的生存关系,生存曲线的生成运用Kaplan-Meier法。结果185例急性白血病患者中64例检测到EBV感染,感染率为34.6%,而对照组仅2例检测到EBV,感染率为5.4%(2/37),急性白血病患者EBV的感染率明显明显高于正常对照组(χ2=12.6,p0.001)。110例ALL病人45例阳性,阳性率40.9%,而75例AML病人19例阳性,阳性率为25.3%,ALL组EBV阳性率明显高于AML组(χ2=4.8,p0.05)。根据流式结果,110例ALL患者中,12例为T-ALL,98例为B-ALL,T-ALL组及B-ALL组EBV感染率分别为8.3%、44.9%,B-ALL组EBV感染率明显高于T-ALL(χ2=4.5,p0.05)。染色体核型分析显示,EBV阳性的B-ALL、T-ALL和AML患者染色体异常率分别为46.3%(25/54)、33.3%(1/3)和30%(9/30),EBV感染和染色体异常之间无明显相关性(χ2=2.3,p0.05)。部分患者的临床随访结果显示:ALL并EBV感染(ALL-EBV+)患者的复发率为56.5%(13/23),而ALL非EBV感染(ALL-EBV-)患者复发率为21.4%(6/28),两组患者的病死率分别为47.8%(11/23)、10.7%(3/28),ALL-EBV+患者的复发率及病死率明显高于ALL-EBV-患者(χ2=6.7、8.7,p0.05)。ALL-EBV+死亡患者的EBV-DNA含量明显高于生存患者(Z=㧟2.3,p0.05)。结论EBV感染可能与急性白血病的发生、发展及免疫分型有关,ALL-EBV+患者复发率高、病死率高,预后不良,提示EBV-DNA检测可作为监测ALL-EBV+患者病情进展的标志。
[Abstract]:Objective to investigate the relationship between Epstein-Barr virus (EBV) infection and the occurrence, development and cytogenetic change of EBV in patients with different types of leukemia (acute lymphoblastic leukemia, acute myeloid leukemia). Methods 185 patients with leukemia were collected as case groups to provide reliable evidence for the close relationship between EBV infection and the occurrence of certain types of leukemia. Of these, 110 were acute lymphoblastic leukemia (ALL) and 75 were acute myeloid leukemia (AMLA), while 37 normal controls ("normal" means that there were no hematological diseases and other cancers of any kind and bone marrow images were generally normal) were collected as controls. The DNA of bone marrow cells was extracted by phenol-chloroform-isoamyl alcohol method, the EBV-DNA in bone marrow cells of all specimens was detected by real-time fluorescence quantitative polymerase chain reaction (FQ-PCR), and the chromosome samples were prepared by short-term culture of bone marrow fluid. The karyotypes were analyzed by means of R banding technique. Immunotyping of ALL was carried out by monoclonal antibody technique, and the correlation between T-ALL and B-ALL and EB virus infection was further analyzed, and some patients were followed up. To further investigate the effect of EB virus infection on the recurrence and death of leukemia patients, Mann-Whitney U nonparametric test method was used to test the quantitative data of two independent samples. The comparison of EBV infection rate among groups was performed by 蠂 2 test and Log-rank test to compare the survival relationship between EBV infection and ALL patients. Kaplan-Meier method was used to generate the survival curve. Results EBV infection was detected in 64 of 185 patients with acute leukemia. The infection rate of EBV in acute leukemia patients was significantly higher than that in the normal control group (蠂 2 / 12. 6 p 0.001 / 110). The positive rate of EBV was 40.9% in 45 patients with ALL and 19 cases in 75 cases of AML. The positive rate of EBV in all group was significantly higher than that in AML group (蠂 2 / 4. 8%, p 0. 05). According to the results of flow cytometry, 12 of 110 ALL patients had EBV infection rate of 8. 3% in T-ALL group and 98 cases in B-ALL group and 8. 3% in B-ALL group respectively (蠂 2 + 4. 5% p0. 05). The karyotype analysis showed that EBV positive rate was significantly higher in BALL group than in T ALL group (蠂 2 / 4. 5% p 0. 05%), and the positive rate of EBV in BALL group was significantly higher than that in T ALL group (蠂 2 / 4. 5% p0. 05%). There was no significant correlation between B-ALL T-ALL infection and chromosome abnormality in B-ALL T-ALL and AML patients (46.3%, 25 / 54 / 33.33%) and 9 / 30 / 30% EBV infection and chromosomal abnormality (蠂 2 / 2.3 p0.05 respectively). The clinical follow-up results of some patients showed that the recurrence rate of patients with EBV infection was 56.535%, while ALL was not EBV infected with ALL-EBV. The recurrence rate and mortality of patients with ALL-EBV were significantly higher than those of patients with ALL-EBV- (蠂 ~ 2 / 6. 7 + 8.7 p 0.05. ALL-EBV). The EBV-DNA content of patients with death of ALL-EBV was significantly higher than that of patients who died of ALL-EBV. The mortality rate of patients with ALL-EBV was significantly higher than that of patients who died of ALL-EBV. The mortality rate of patients with ALL-EBV was significantly higher than that of patients who died of ALL-EBV. Conclusion EBV infection may be associated with the occurrence, development and immunological classification of ALL-EBV patients with high recurrence rate, high fatality rate and poor prognosis, suggesting that EBV-DNA detection can be used as a marker to monitor the progression of ALL-EBV patients.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R733.71

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