采用芯片数字PCR技术检测造血干细胞移植后儿童人巨细胞病毒感染的病毒载量

The Viral Load of Human Cytomegalovirus Infection in Children following Hematopoietic Stem Cell Transplant by Chip Digital PCR.

作者信息

Wang Wen-Jun, Feng Miao, He Feng, Song Juan, Song Qin-Qin, Xia Dong, Liu Rong, Yao Hai-Lan, Han Jun

机构信息

State Key Laboratory of Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, 155 Changbai Rd, Beijing 102206, China.

Department of Biochemistry & Immunology, Capital Institute of Pediatrics, YaBao Rd, Beijing 100020, China.

出版信息

Can J Infect Dis Med Microbiol. 2022 Oct 17;2022:2786841. doi: 10.1155/2022/2786841. eCollection 2022.

DOI:10.1155/2022/2786841

PMID:36300166

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9592232/

Abstract

OBJECTIVE

To detect viral load in human cytomegalovirus (HCMV) infection children after hematopoietic stem cell transplant (HSCT) by chip digital PCR (cdPCR).

METHODS

The plasmid pUC57-UL83 containing the HCMV-UL83 gene and HCMV AD169 strain were used to evaluate the sensitivity of cdPCR. Either HSV-1, HSV-2, VZV, EBV, HHV-6, or HHV-7 was used to evaluate the specificity of HCMV cdPCR. The cdPCR was compared with quantitative PCR (qPCR) by detecting HCMV infection in 125 children's whole blood samples following HSCT.

RESULTS

The limit of detection (LOD) of HCMV cdPCR was 103 copies/ml and the qPCR LOD was 297 copies/ml for plasmid pUC57-UL83. The result of HCMV cdPCR was 146 copies/ml for the HCMV AD169 strain, indicating that the sensitivity of cdPCR was higher than that of qPCR. There is no cross-reaction between HCMV cdPCR and other herpes viruses. The incidence of HCMV infection was 30.40% in 125 children following HSCT by cdPCR. The range of the HCMV viral load was from 107 copies/ml to 6600 copies/ml by cdPCR.

CONCLUSIONS

cdPCR is more sensitive than qPCR for detecting HCMV viral load. Furthermore, the cdPCR could be used to detect the viral load of HCMV infection before or after HSCT in children.

摘要

目的

采用芯片数字PCR（cdPCR）检测造血干细胞移植（HSCT）后人类巨细胞病毒（HCMV）感染儿童的病毒载量。

方法

使用含有HCMV-UL83基因的质粒pUC57-UL83和HCMV AD169株评估cdPCR的灵敏度。使用单纯疱疹病毒1型（HSV-1）、单纯疱疹病毒2型（HSV-2）、水痘带状疱疹病毒（VZV）、EB病毒（EBV）、人疱疹病毒6型（HHV-6）或人疱疹病毒7型（HHV-7）评估HCMV cdPCR的特异性。通过检测125例HSCT后儿童全血样本中的HCMV感染情况，将cdPCR与定量PCR（qPCR）进行比较。

结果

对于质粒pUC57-UL83，HCMV cdPCR的检测限（LOD）为103拷贝/毫升，qPCR的LOD为297拷贝/毫升。HCMV AD169株的HCMV cdPCR结果为146拷贝/毫升，表明cdPCR的灵敏度高于qPCR。HCMV cdPCR与其他疱疹病毒之间无交叉反应。通过cdPCR检测，125例HSCT后儿童中HCMV感染的发生率为30.40%。通过cdPCR检测，HCMV病毒载量范围为107拷贝/毫升至6600拷贝/毫升。