异基因造血干细胞移植后实时定量聚合酶链反应在巨细胞病毒感染诊断和治疗中的应用

目的 探讨异基因造血干细胞移植(allo-HSCT)后应用实时定量聚合酶链反应(RQ-PCR)技术检测血浆巨细胞病毒(CMV)DNA载量,诊断CMV感染及指导临床抗CMV治疗的意义.方法 统以318例接受allo-HSCT患者为研究对象,其中同胞HLA相合HSCT患者160例,血缘关系配型不合127例,非血缘关系移植31例.移植前供受者采用ELISA方法 进行CMV血清学抗体检测.移植后采用RQ-PCR方法 检测血浆CMV DNA,6×102拷贝数/ml为CMV PCR阳性.全部患者在预处理阶段-9 d到-2 d应用更昔洛韦预防CMV感染.抢先治疗主要应用更昔洛韦或膦甲酸钠,或两药联合治疗.结果 移植后100天(+100 d)内CMV感染累计发生率40.6%,同胞相合、血缘关系配型不合及非血缘关系移植三种移植方式,+100 d内CMV感染累计发生率分别为17.5%、66.1%、45.2%.无论是单因素还是多因素分析均表明,血缘关系配型不合移植和非血缘关系移植,预处理方案中使用抗胸腺细胞球蛋白(ATG)以及中重度急性移植物抗宿主病是导致CMV感染发生的危险因素.在早期应用抢先治疗后,总体+100 d内CMV病累计发生率为8.8%.三种移植方式+100 d内CMV病累计发生率分别为5.6%、9.4%、22.6%.且三种移植方式CMV感染患者2年生存率差异无统计学意义.结论 allo-HSCT后RQ-PCR检测血浆CMV DNA可有效用于CMV感染诊断和监测,并有利于指导临床抗CMV治疗,减少移植后CMV感染高危患者CMV疾病的发生.

Application of real time polymerase chain reaction to the diagnosis and treatment of cytomegalovirus infection after anogeneic hematopoietic stem cell transplantation

Objective To investigate the role of real time quantitative polymerase chain reaction (RQ-PCR) in the diagnosis and treatment of recipients cytomegalovirus (CMV) infection after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods 318 patients received alIo-HSCT were stud-ied. 160 patients received transplants from HLA matched sibling donors; 127 from HLA mismatched related donors; 31 from unrelated donors. Before transplant recipients and donors received CMV serological test by ELISA. After transplant RQ-PCR was used to test and monitor CMV-DNA in plasma of patients. A positive CMV-PCR was defined as >6 x 102 copies/ml. Ganciclovir was used for CMV prophylaxis in all patients at -9 d to-2 d of conditioning regimen period. Ganciclovir, foscarnet, or combination of the two drugs were used as the preemptive therapy. Results The total 100-day cumulative incidence of CMV infection was 40.6%. The incidence was 17.5%, 66.1% and 45.2% for the HLA matched sibling, HLA mismatched re-lated(MMR) and unrelated donor(MUR) HSCT respectively. Multivariate analysis showed MMR HSCT, MUR HSCT, ATG containing preparative regimen and moderate to severe aGVHD were the risk factors for CMV infection after HSCT. The 100 day cumulative incidence of CMV disease was 8.8% and 5.6% ,9.4%, 22.6% respectively for total and three kinds of HSCT after early preemptive therapy. Two-year survival of CMV infection was similar in the three kinds of SCT. Conclusion Detection of CMV DNA in plasma by real time PCR appears to be effective for the diagnosis and surveillance of CMV infection after HSCT. It may help to initiate antiviral therapy and reduce the incidence of CMV disease in the patients with high risk of CMV in-fection.