ddPCR在儿童异基因造血干细胞移植后EB病毒感染监测中的初步应用

Exploratory use of ddPCR in monitoring Epstein-Barr virus infection following allogeneic hematopoietic stem cell transplantation in children

目的本研究应用微滴式数字PCR(droplet digital PCR,ddPCR)检测儿童异基因造血干细胞移植术(allogeneic hematopoietic stem cell transplantation,alloHSCT)术后EB病毒(Epstein-Barr virus,EBV)感染的情况,初步探讨其在临床中作为alloHSCT移植术后EBV感染监测的补充检测方法的可行性。方法收集47例alloHSCT术后儿童共290份外周血样本,使用ddPCR和实时定量PCR(real-time quantitative PCR,qPCR)分别检测血浆中EBV DNA载量,比较两者的检出效能。通过对其中39例患儿进行连续检测,观察EBV DNA载量的动态变化,并分析两种方法的优劣势。结果ddPCR的EBV阳性检出率显著高于qPCR(χ^(2)=20.25,P<0.001),尤其在低病毒载量样本中表现更佳。在连续监测EBV DNA期间,39例患儿中有14例出现ddPCR阳性结果,其中有2例在ddPCR检测阳性、qPCR检测阴性的同时出现了皮疹和发热,提示ddPCR在早期发现EBV感染方面更为敏感。结论ddPCR在alloHSCT术后患儿EBV感染监测中具有一定优势,尤其是对于低病毒载量的样本。然而,由于该技术在临床应用中尚处于探索阶段,仍需更多的研究和实践来证实ddPCR的应用价值。

Objective This study aimed to introduce droplet digital PCR(ddPCR)for monitoring Epstein-Barr virus(EBV)infection after allogeneic hematopoietic stem cell transplantation(alloHSCT)in children and assess its viability as a complementary detection method in clinical settings.Methods A total of 290 blood samples from 47 children undergoing alloHSCT were collected.Both ddPCR and real-time quantitative PCR(qPCR)were employed to detect EBV DNA load in plasma,with a comparison of detection efficiencies between the two methods.Continuous monitoring of 39 children was conducted to observe dynamic changes in EBV DNA load in plasma and analyze the merits and drawbacks of both methods.Results The EBV positive detection rate of ddPCR was significantly higher than that of qPCR(χ^(2)=20.25,P<0.001),particularly in samples with low viral loads.Among the children monitored continuously for EBV DNA,14 out of 39 exhibited positive ddPCR result.Notably,in two cases where patients displayed rash and fever symptoms with positive ddPCR result but negative qPCR result,ddPCR demonstrated heightened sensitivity in early EBV infection detection.Conclusions ddPCR holds certain advantages in monitoring EBV infection post-alloHSCT in children,especially for samples with low viral loads.However,as this method is still in the exploratory stage of clinical application,further research and practice are needed to validate its utility.