摘要
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 establish a convenient and specific molecular method for cytomegalovirus （CMV） surveillance in recipients of allogeneic hematopoietic stem cell transplantation （allo-HSCT） and analyze the correlation between cytomegalovirus genotypes and transplant-related complication to decrease CMV- related mortality through restricted endoenzyme digestion. Methods 135 blood .samples were regularly collected from 79 consecutive patients who received allo-HSCT between April 2001 and April 2003. Of the 79 patients observed, median age was 32 （range from 11 to 60） and the ratio of male and female was 1.63： 1. DNA extracted from whole blood was amplified with CMV sequence using Nested PCR and gB genotypes were identified through restriction enzyme analysis. Results Among the 135 clinical isolates from 79 patients, 42 cases （66 specimens） were Positive, The gB genotype from 42 cases （45 samples） were identified as gB1, 21 （46.7 %）; gB2, 14 （31.1%）; gB3, 7 （15.6 %）; gB4, 3 （6.7%）, respectively, by restriction enzyme analysis. Among them, 3 patients were successively infected with CMV gB1 and gB2. The incidence of GVHD in groups of CMV（ ＋ ） and CMV（-） was 81.0% and 32.4%, respectively （P〈0.01）. Incidence of aGVHD Ⅱ～Ⅳ/cGVHD in different CMVgB types was as follows： gB1, 81.0 %; gB2, 50.0%; gB3, 42.9% as well as gB4, 0, respectively. Conclusions Nested-PCR to amplify CMV gB gene is a convenient and speacific molecular method for cytomegalovirus surveillance. Close correlation was presented between CMV gB（ ＋ ） and GVHD. CMV gB1 and gB2 were leading genotypes isolated from our Center. CMV gB1 infection was particularly associated with significantly high incidence of GVHD. This CMV nested-PCR assay can give doctors some valuable prognostic message to reduce transplant-related mortality.
出处
《中华器官移植杂志》
CAS
CSCD
北大核心
2006年第6期348-351,共4页
Chinese Journal of Organ Transplantation
基金
全军医学科研"九五"计划项目基金资助(98M130)
关键词
造血干细胞移植
巨细胞病毒
聚合酶链反应
Hematopoietic stem cell transplantation
Cytomegalovirus
Polymerase chain reaction
