發(fā)布時間：2018-03-20 21:34

  本文選題：EB病毒　切入點：實時熒光定量聚合酶鏈式反應　出處：《青島大學》2017年碩士論文　論文類型：學位論文

【摘要】：目的檢測不同類型白血病(急性淋巴細胞白血病、急性髓細胞白血病)患者EB病毒(EBV)的感染情況,以探討EBV與白血病的發(fā)生、發(fā)展及細胞遺傳學改變的相關性,為揭示EBV感染與某些類型白血病的發(fā)生密切相關提供可靠依據(jù)。方法收集185例白血病患者作為病例組,其中110例為急性淋巴細胞白血病(ALL),75例為急性髓細胞白血病(AML),同時收集37例正常人(“正�！敝笡]有任何類型的血液系統(tǒng)疾病和其他癌癥且骨髓象大致正常)作為對照組,采用常規(guī)苯酚-氯仿-異戊醇方法提取骨髓細胞DNA,采用實時熒光定量聚合酶鏈反應(FQ-PCR)檢測所有標本骨髓細胞中EBV-DNA;采用骨髓液短期培養(yǎng)法制備染色體標本并采用常規(guī)R顯帶技術(shù)分析核型;運用單克隆抗體技術(shù)對ALL進行免疫分型,進一步分析T-ALL和B-ALL與EB病毒感染的相關性,并對部分患者進行了臨床隨訪,以進一步探討EB病毒感染對白血病患者的復發(fā)及死亡的影響。兩獨立樣本定量資料的檢驗應用Mann-Whitney U非參數(shù)檢驗方法;各組間EBV感染率的比較采用χ2檢驗;Log-rank檢驗用于比較EBV感染與ALL患者之間的生存關系,生存曲線的生成運用Kaplan-Meier法。結(jié)果185例急性白血病患者中64例檢測到EBV感染,感染率為34.6%,而對照組僅2例檢測到EBV,感染率為5.4%(2/37),急性白血病患者EBV的感染率明顯明顯高于正常對照組(χ2=12.6,p0.001)。110例ALL病人45例陽性,陽性率40.9%,而75例AML病人19例陽性,陽性率為25.3%,ALL組EBV陽性率明顯高于AML組(χ2=4.8,p0.05)。根據(jù)流式結(jié)果,110例ALL患者中,12例為T-ALL,98例為B-ALL,T-ALL組及B-ALL組EBV感染率分別為8.3%、44.9%,B-ALL組EBV感染率明顯高于T-ALL(χ2=4.5,p0.05)。染色體核型分析顯示,EBV陽性的B-ALL、T-ALL和AML患者染色體異常率分別為46.3%(25/54)、33.3%(1/3)和30%(9/30),EBV感染和染色體異常之間無明顯相關性(χ2=2.3,p0.05)。部分患者的臨床隨訪結(jié)果顯示:ALL并EBV感染(ALL-EBV+)患者的復發(fā)率為56.5%(13/23),而ALL非EBV感染(ALL-EBV-)患者復發(fā)率為21.4%(6/28),兩組患者的病死率分別為47.8%(11/23)、10.7%(3/28),ALL-EBV+患者的復發(fā)率及病死率明顯高于ALL-EBV-患者(χ2=6.7、8.7,p0.05)。ALL-EBV+死亡患者的EBV-DNA含量明顯高于生存患者(Z=㧟2.3,p0.05)。結(jié)論EBV感染可能與急性白血病的發(fā)生、發(fā)展及免疫分型有關,ALL-EBV+患者復發(fā)率高、病死率高,預后不良,提示EBV-DNA檢測可作為監(jiān)測ALL-EBV+患者病情進展的標志。
[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.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R733.71

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