Rogers B B, Conlin C, Timmons C F, Dawson D B, Krisher K, Andrews W S
Department of Pathology, Children's Medical Center, Dallas, TX 75235, USA.
Pediatr Pathol Lab Med. 1997 May-Jun;17(3):391-400.
Epstein-Barr virus (EBV)-associated illnesses in posttransplant patients are difficult to diagnose. Attempts to aid in the diagnosis of such illnesses using the polymerase chain reaction (PCR) analysis for EBV have met with variable success due to the potential exquisite sensitivity of the assay. We have designed a relatively insensitive EBV PCR assay and compared the results with objective evidence of EBV activity including serologic response and in situ hybridization for the EBV genome. Eighty-five specimens from 65 patients were analyzed by the EBV PCR using DNA from whole blood. EBV serologic evaluation was done on 53 of the samples and in situ hybridization for EBV (EBER-1 mRNA) on 46 paired liver biopsies. Of 85 samples, 25 (29%) were positive for EBV using the PCR assay. Intensity of amplification was graded 0.5-1+ (weak) to 3+ (strong). Using these criteria, 19 EBV PCR-positive samples were graded 0.5-1+, 5 were graded 2+, and 1 was graded 3+. Of the moderate to strongly positive samples (2+ or 3+), five of six had two or more EBER-1-positive cells in the liver biopsies. Of the remaining 40 liver biopsies with either negative or weak positive PCR results, 3 had only single cells positive for EBER-1; the remainder were negative. In addition, PCR-positive results correlated with increasing EBV anti-early antigen antibody (P = .005) and viral capsid antigen IgG immunoglobulin G VCA (P = .05) EBV-positive results using the PCR assay correlated with objective evidence for increased EBV burden in children after liver transplantation. These preliminary data suggest that this PCR test may be useful to help guide immunosuppressive therapy in the posttransplant patient. Further evaluation using larger numbers of patients will be necessary to confirm this.
移植后患者中与爱泼斯坦-巴尔病毒(EBV)相关的疾病难以诊断。由于聚合酶链反应(PCR)分析检测EBV的潜在高灵敏度,利用该方法辅助诊断此类疾病的尝试取得的成功参差不齐。我们设计了一种相对不敏感的EBV PCR检测方法,并将结果与EBV活性的客观证据进行比较,包括血清学反应和EBV基因组的原位杂交。使用全血DNA,通过EBV PCR对65例患者的85份标本进行了分析。对其中53份样本进行了EBV血清学评估,对46对肝活检组织进行了EBV(EBER-1 mRNA)原位杂交。在85份样本中,25份(29%)通过PCR检测为EBV阳性。扩增强度分为0.5 - 1+(弱)至3+(强)。根据这些标准,19份EBV PCR阳性样本评分为0.5 - 1+,5份评分为2+,1份评分为3+。在中度至强阳性样本(2+或3+)中,六分之五的肝活检组织中有两个或更多EBER-1阳性细胞。在其余40份PCR结果为阴性或弱阳性的肝活检组织中,3份仅有单个细胞EBER-1阳性;其余均为阴性。此外,PCR阳性结果与EBV抗早期抗原抗体增加相关(P = 0.005),与病毒衣壳抗原IgG(VCA)相关(P = 0.05)。肝移植后儿童中,PCR检测的EBV阳性结果与EBV负荷增加的客观证据相关。这些初步数据表明,这种PCR检测可能有助于指导移植后患者的免疫抑制治疗。需要使用更多患者进行进一步评估以证实这一点。