Experimental and clinical transplantation : official journal of the Middle East Society for Organ Transplantation · 2021
Sequencing of Cell-Free DNA to Monitor Cytomegalovirus Infection After Liver Transplant.
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Li Jiequn, Tan Jieqiong, Chen Guangshun, Li Qiang, Zhou Zhengjun, Bin Yangyang, Qi Haizhi, Si Zhongzhou
Аннотация
OBJECTIVES: In the present study, we investigated donor-derived cell-free DNA dynamics and assessed the diagnostic efficacy of 2 tests: the sequencing of cytomegalovirus-derived cell-free DNA and the quantitative nucleic acid amplification test in cytomegalovirus infection following liver transplant.
MATERIALS AND METHODS: We first examined 6 patients who were identified with active cytomegalovirus DNAemia by both quantitative nucleic acid amp-lification test and next-generation sequencing of cytomegalovirus-derived cell-free DNA and then performed a receiver operating characteristic analysis to evaluate the efficacy of cell-free DNA sequencing and establish a cutoff for this assay. Further validation of the next-generation sequencing method was also performed in 84 liver transplant recipients. The study protocol conformed to the ethical guidelines of the Declaration of Helsinki and the Declaration of Istanbul.
RESULTS: In the first 6 patients, there was no significant correlation between the cytomegalovirus infection and donor-derived cell-free DNA. We determined that the levels of cytomegalovirus-derived cell-free DNA sequencing directly correlate with the results of the quantitative nucleic acid amplification test (area under the curve 0.982) and obtained a value of 0.015% as a cutoff for the cell-free DNA sequencing assay. In the validation cohort composed of 84 liver transplant recipients, next-generation sequencing of cell-free DNA revealed the occurrence of cytomegalovirus infection that remains otherwise undetected by the quantitative nucleic acid amplification test.
CONCLUSIONS: Cytomegalovirus infections that do not cause direct graft injury (cytomegalovirus-related hepatitis) did not result in elevations of donor-derived cell-free DNA. Next-generation sequencing of cytomegalovirus-derived cell-free DNA provides a potential tool for detection of cytomegalovirus infection that remains undetected by the quantitative nucleic acid amplification test.
MATERIALS AND METHODS: We first examined 6 patients who were identified with active cytomegalovirus DNAemia by both quantitative nucleic acid amp-lification test and next-generation sequencing of cytomegalovirus-derived cell-free DNA and then performed a receiver operating characteristic analysis to evaluate the efficacy of cell-free DNA sequencing and establish a cutoff for this assay. Further validation of the next-generation sequencing method was also performed in 84 liver transplant recipients. The study protocol conformed to the ethical guidelines of the Declaration of Helsinki and the Declaration of Istanbul.
RESULTS: In the first 6 patients, there was no significant correlation between the cytomegalovirus infection and donor-derived cell-free DNA. We determined that the levels of cytomegalovirus-derived cell-free DNA sequencing directly correlate with the results of the quantitative nucleic acid amplification test (area under the curve 0.982) and obtained a value of 0.015% as a cutoff for the cell-free DNA sequencing assay. In the validation cohort composed of 84 liver transplant recipients, next-generation sequencing of cell-free DNA revealed the occurrence of cytomegalovirus infection that remains otherwise undetected by the quantitative nucleic acid amplification test.
CONCLUSIONS: Cytomegalovirus infections that do not cause direct graft injury (cytomegalovirus-related hepatitis) did not result in elevations of donor-derived cell-free DNA. Next-generation sequencing of cytomegalovirus-derived cell-free DNA provides a potential tool for detection of cytomegalovirus infection that remains undetected by the quantitative nucleic acid amplification test.