State Key Laboratory of Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
Department of Hematology, Children's Hospital of Capital Institute of Pediatrics, Beijing 100020, China.
Biomed Environ Sci. 2022 Sep 20;35(9):804-810. doi: 10.3967/bes2022.052.
To detect the Epstein-Barr virus (EBV) viral load of children after hematopoietic stem cell transplantation (HSCT) using chip digital PCR (cdPCR).
The sensitivity of cdPCR was determined using EBV plasmids and the EBV B95-8 strain. The specificity of EBV cdPCR was evaluated using the EBV B95-8 strain and other herpesviruses (herpes simplex virus 1, herpes simplex virus 2, varicella zoster virus, human cytomegalovirus, human herpesvirus 6, and human herpesvirus 7). From May 2019 to September 2020, 64 serum samples of children following HSCT were collected. EBV infection and the viral load of serum samples were detected by cdPCR. The epidemiological characteristics of EBV infections were analyzed in HSCT patients.
The limit of detection of EBV cdPCR was 110 copies/mL, and the limit of detection of EBV quantitative PCR was 327 copies/mL for the pUC57-BALF5 plasmid. The result of EBV cdPCR was up to 121 copies/mL in the EBV B95-8 strain, and both were more sensitive than that of quantitative PCR. Using cdPCR, the incidence of EBV infection was 18.75% in 64 children after HSCT. The minimum EBV viral load was 140 copies/mL, and the maximum viral load was 3,209 copies/mL using cdPCR. The average hospital stay of children with EBV infection (184 ± 91 days) was longer than that of children without EBV infection (125 ± 79 days), = 0.026.
EBV cdPCR had good sensitivity and specificity. The incidence of EBV infection was 18.75% in 64 children after HSCT from May 2019 to September 2020. EBV cdPCR could therefore be a novel method to detect EBV viral load in children after HSCT.
使用芯片数字 PCR（cdPCR）检测造血干细胞移植（HSCT）后儿童的 EBV 病毒（EBV）载量。
使用 EBV 质粒和 EBV B95-8 株确定 cdPCR 的灵敏度。使用 EBV B95-8 株和其他疱疹病毒（单纯疱疹病毒 1、单纯疱疹病毒 2、水痘带状疱疹病毒、人巨细胞病毒、人疱疹病毒 6、人疱疹病毒 7）评估 EBV cdPCR 的特异性。2019 年 5 月至 2020 年 9 月，采集 64 例 HSCT 后儿童的血清样本。通过 cdPCR 检测 EBV 感染和血清样本的病毒载量。分析 HSCT 患者 EBV 感染的流行病学特征。
EBV cdPCR 的检测下限为 110 拷贝/mL，pUC57-BALF5 质粒的 EBV 定量 PCR 检测下限为 327 拷贝/mL。EBV B95-8 株的 EBV cdPCR 结果高达 121 拷贝/mL，均比定量 PCR 更敏感。通过 cdPCR，64 例 HSCT 后儿童的 EBV 感染发生率为 18.75%。EBV 病毒载量最低为 140 拷贝/mL，最高为 3209 拷贝/mL。EBV 感染患儿的平均住院时间（184 ± 91 天）长于无 EBV 感染患儿（125 ± 79 天）， = 0.026。
EBV cdPCR 具有良好的灵敏度和特异性。2019 年 5 月至 2020 年 9 月，64 例 HSCT 后儿童 EBV 感染发生率为 18.75%。因此，EBV cdPCR 可能是一种检测 HSCT 后儿童 EBV 病毒载量的新方法。
