肝肾移植术后受者人巨细胞病毒两种检测方法的对比研究

COMPARATIVE STUDY OF TWO METHODS TO DETECT HUMAN CYTOMEGALOVIRUS IN HEPATIC OR RENAL TRANSPLANT RECIPIENTS

目的:探讨巢式PCR(Nest PCR)检测器官移植术后受体人巨细胞病毒(HCMV)DNA,并与传统ELISA法作方法学对照。方法:巢式PCR针对HCMV AD169株IEA基因设计内外两对引物,对59例肝、肾移植术后受体(肝移植27例、肾移植32例)血、尿标本进行HCMV DNA检测。分离血清作EIASA检测IgG、IgM。结果:血液Nest PCR阳性率肝移植62.9％,肾移植46.8％;ELISA法阳性率:IgC35.5％,IgM27.1％,IgG+IgM18.6％,IgM阳性标本(16例)DNA检测皆为阳性,6例IgG、IgM皆阴性标本DNA检测仍为阳性。两法检测,P<0.05,证明两法存在显著差异,结论:巢式PCR是一种敏感特异、简便快速诊断HCMV感染的方法,灵敏度及特异性高于传统ELISA,能弥补ELISA的假阴性,更适用于临床检测器官移植术后HCMV感染。

Objective: To compare nest polymerase chain reaction (Nest PCR) with enzyme - linked immunoadsordent assay (ELISA) in detecing DNA in human cytomegalovirus (HCMV) in patients receiving organ transplants. Methos: Based on two pairs of internal and external primers in IEA gene from HCMV AD169 strain, Nest PCR probe was designed, and blood and urine samples from 59 cases receiving hepatic (27 cases) or renal (32 cases) transplants were detected with the probe. At the same time, the levels of IgG and IgM in sear of these subjects were detected with ELISA. Result: With Nest PCR, positive rates of HCMV in blood samples from hepatic and renal transplant recipients were 62. 9% and 46. 8, respectively. With ELISA, the positive rate of IgG, IgM and IgG+ IgM were 35. 5%, 27. 1% and 18. 6%, respectively. The DNA detection of 16 cases with positive IgM was negative, but that of 6 cases with negative IgG and/or IgM was postive. The difference between both methods was significant ( p < 0. 05). Conclusion: Nest PCR is a sensitive, specific, simple and rapid method to diagnose HCMV infection, and its sensitivity and particularity overmatch ELISA. Therefore, it is suggested that Nest PCR is more suitable to detect HCMV infection after organ transplant in clinical than ELISA.