Background: Donor-derived cell free DNA (ddcf DNA) has been reported as a universal noninvasive biomarker for rejection monitoring in heart, kidney, liver, and lung transplantation. Current approaches based on next-ge...Background: Donor-derived cell free DNA (ddcf DNA) has been reported as a universal noninvasive biomarker for rejection monitoring in heart, kidney, liver, and lung transplantation. Current approaches based on next-generation sequencing for quantification of ddcf DNA, although promising, may be restricted by the requirement for donor material, as donor samples may not be available. Methods: We proposed a novel next-generation sequencing approach without donor-derived material and compared the non-donor-derived approach and the donor-derived approach using simulation testing and 69 clinical specimens. We also evaluated the performance for acute rejection and infection monitoring in lung transplantation. Results: The non-donor-derived approach reached similar efficacy as the donor-derived approach with a significant linear correlation of R2 = 0.98. Subsequent validation in clinical specimens demonstrated significant difference between the acute rejection group (4.83% ± 2.11%, mean ± SD) and the non-rejection group (1.61% ± 0.63%, mean ± SD) (P ’s t test). With the cut-off value of 2.999, our approach had 90.48% sensitivity (95% CI, 69.62% - 98.83%), 100% specificity (95% CI, 91.59% - 100%), and AUC 0.9266 (95% CI, 0.8277 - 1.026). The test also had the ability to simultaneously detect infectious agents, especially cytomegalovirus, as compared with the clinical test. Conclusion: The proposed approach without donor-derived material could potentially be used to monitor acute rejection and infection in lung transplantation and may be applied to other types of solid organ transplantation.展开更多
目的:探讨影响万古霉素、哌拉西林/他唑巴坦、美罗培南3种药物治疗药物监测(therapeutic drug monitoring,TDM)靶值和药动学/药效学(pharmacokinetic/pharmacodynamic,PK/PD)靶值未达标的危险因素以及2种靶值和微生物疗效的关系,为精准...目的:探讨影响万古霉素、哌拉西林/他唑巴坦、美罗培南3种药物治疗药物监测(therapeutic drug monitoring,TDM)靶值和药动学/药效学(pharmacokinetic/pharmacodynamic,PK/PD)靶值未达标的危险因素以及2种靶值和微生物疗效的关系,为精准用药提供科学依据。方法:纳入2021年3月至2022年3月接受万古霉素、哌拉西林/他唑巴坦、美罗培南中至少1种抗菌药物联合治疗的危重症患者。利用优化采样方案采集每位患者的3~4份血浆样本,借助非房室模型结合经典药动学理论计算PK/PD参数。采用多因素分析探究3种药物TDM靶值和PK/PD靶值未达标的相关危险因素。结果:共纳入接受万古霉素、哌拉西林/他唑巴坦或美罗培南治疗的25名危重症患者的96份血浆样本,共计151份血药浓度-时间数据。万古霉素、哌拉西林/他唑巴坦和美罗培南的谷浓度(trough concentration,Cmin)达标率低,分别为20.00%、22.22%和23.53%;万古霉素PK/PD靶值达标率、未达标率和超标率分别为20.00%、60.00%和20.00%。哌拉西林/他唑巴坦和美罗培南的PK/PD靶值达标率分别为11.11%和17.65%,未达标率分别高达88.89%和76.47%。影响因素探究发现肌酐清除率是影响3种抗菌药物Cmin和PK/PD靶值不达标的独立危险因素。细菌根除组患者的万古霉素Cmin[(15.329±7.522)mg·L^(-1)vs.(6.273±0.754)mg·L^(-1),P=0.019]和AUC0~24 h/MIC(578.47±413.86 vs.175.94±17.07,P=0.028)均显著高于细菌未根除组。结论:万古霉素、哌拉西林/他唑巴坦、美罗培南在危重症患者中的TDM靶值和PK/PD靶值达标率低,未达标率高,普遍存在低暴露和过暴露情况。提高TDM靶值和PK/PD靶值达标率均可能提高患者细菌根除率,因此十分有必要在此类患者中开展TDM,为精准给药提供参考。展开更多
文摘Background: Donor-derived cell free DNA (ddcf DNA) has been reported as a universal noninvasive biomarker for rejection monitoring in heart, kidney, liver, and lung transplantation. Current approaches based on next-generation sequencing for quantification of ddcf DNA, although promising, may be restricted by the requirement for donor material, as donor samples may not be available. Methods: We proposed a novel next-generation sequencing approach without donor-derived material and compared the non-donor-derived approach and the donor-derived approach using simulation testing and 69 clinical specimens. We also evaluated the performance for acute rejection and infection monitoring in lung transplantation. Results: The non-donor-derived approach reached similar efficacy as the donor-derived approach with a significant linear correlation of R2 = 0.98. Subsequent validation in clinical specimens demonstrated significant difference between the acute rejection group (4.83% ± 2.11%, mean ± SD) and the non-rejection group (1.61% ± 0.63%, mean ± SD) (P ’s t test). With the cut-off value of 2.999, our approach had 90.48% sensitivity (95% CI, 69.62% - 98.83%), 100% specificity (95% CI, 91.59% - 100%), and AUC 0.9266 (95% CI, 0.8277 - 1.026). The test also had the ability to simultaneously detect infectious agents, especially cytomegalovirus, as compared with the clinical test. Conclusion: The proposed approach without donor-derived material could potentially be used to monitor acute rejection and infection in lung transplantation and may be applied to other types of solid organ transplantation.
