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The Viral Load of Human Cytomegalovirus Infection in Children following Hematopoietic Stem Cell Transplant by Chip
Wen-Jun Wang1, Miao Feng2, Feng He2
1State 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.
Insights
Chip digital PCR (cdPCR) offers higher sensitivity for detecting human cytomegalovirus (HCMV) viral load in children post-hematopoietic stem cell transplant (HSCT) compared to quantitative PCR (qPCR). This advanced method aids in monitoring HCMV infection effectively.
Area of Science:
- Virology
- Molecular Diagnostics
- Transplantation Immunology
Background:
- Human cytomegalovirus (HCMV) poses a significant risk to immunocompromised patients, particularly children undergoing hematopoietic stem cell transplant (HSCT).
- Accurate and sensitive viral load detection is crucial for managing HCMV infection post-HSCT.
Purpose of the Study:
- To evaluate the sensitivity and specificity of chip digital PCR (cdPCR) for detecting HCMV viral load in children after HSCT.
- To compare the performance of cdPCR with quantitative PCR (qPCR) in this patient population.
Main Methods:
- The study utilized plasmid pUC57-UL83 and HCMV AD169 strain to assess cdPCR sensitivity.
- Specificity was determined using a panel of other herpesviruses (HSV-1, HSV-2, VZV, EBV, HHV-6, HHV-7).
- cdPCR was compared against qPCR using whole blood samples from 125 children post-HSCT.
Main Results:
- cdPCR demonstrated a lower limit of detection (LOD) of 103 copies/ml compared to qPCR's 297 copies/ml for plasmid pUC57-UL83.
- cdPCR showed higher sensitivity with the HCMV AD169 strain (146 copies/ml) than qPCR.
- HCMV infection was detected in 30.40% of children, with viral loads ranging from 107 to 6600 copies/ml by cdPCR. No cross-reactivity was observed with other herpesviruses.
Conclusions:
- cdPCR is a more sensitive method than qPCR for quantifying HCMV viral load.
- cdPCR is a viable tool for monitoring HCMV infection in children before or after HSCT.
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.
