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目的为了降低异基因造血干细胞移植(allo-HSCT)后巨细胞病毒(CMV)感染的相关死亡率,寻求一种更快捷、更特异的诊断CMV感染的分子生物学方法。方法2001年4月至2003年4月,从79例接受异基因造血干细胞移植的患者中采集了135份外周血标本。采用巢式聚合酶链反应(nested-PCR)方法检测患者外周血中CMVgB DNA,阳性标本做酶切分型,部分行序列测定。结果CMVgB DNA检测中有42例患者(53.9%)的66份标本(48.9%)阳性;对其中阳性患者的45份标本进行了酶切分型,结果CMV gB1型21例(46.7%),CMV gB2型14例(31.1%),CMV gB3型7例(15.6%),CMV gB4型3例(6.7%)。先后有2种型别的CMV感染者有3例(3.8%)。经分析,CMV gB阳性和CMV gB阴性患者GVHD的发生率分别为81.0%和32.4%(P<0.01);CMV gB1、gB2、gB3以及gB4型II~IV度急性GVHD及慢性GVHD的发生率分别为:81.0%、50.0%、42.9%和0。结论Nested-PCR方法可快捷特异地检测CMV感染。CMVgB1、2型中,重度急性GVHD和慢性GVHD发生率较高。CMV gB基因分型检测可有效地指导临床抗病毒治疗,且对移植后患者的临床转归有一定的预测价值。
Objective To seek a more rapid and specific molecular biological method to diagnose CMV infection in order to reduce the related mortality of cytomegalovirus (CMV) infection after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods From April 2001 to April 2003, 135 peripheral blood samples were collected from 79 patients who underwent allogeneic hematopoietic stem cell transplantation. The nested-PCR method was used to detect CMVgB DNA in peripheral blood of patients. The positive samples were digested by restriction enzyme digestion and sequenced. Results In the CMVgB DNA test, 66 samples (48.9%) of 42 patients (53.9%) were positive. 45 of the positive samples were digested by restriction enzyme digestion. The results showed that CMV gB1 was found in 21 cases (46.7%), CMV There were 14 cases (31.1%) of gB2, 7 cases (15.6%) of CMV gB3 and 3 cases (6.7%) of CMV gB4. There are three types of CMV infection (3.8%). The incidences of GVHD in CMV gB positive and CMV gB negative patients were 81.0% and 32.4%, respectively (P <0.01). The incidences of acute GVHD and chronic GVHD in CMV gB1, gB2, gB3 and gB4 type II to IV were respectively As follows: 81.0%, 50.0%, 42.9% and 0. Conclusion Nested-PCR method can detect CMV infection rapidly and specifically. CMVgB1, 2 type, severe acute GVHD and chronic GVHD higher incidence. CMV gB genotyping test can effectively guide the clinical anti-virus therapy, and the prognosis of patients after transplantation has some predictive value.