Construction and evaluation of a viral multiplex detection method based on ddPCR for kidney transplantation
10.3760/cma.j.cn114452-20240723-00396
- VernacularTitle:基于ddPCR的肾移植术后病毒多联检方法构建及评价
- Author:
Shuangshuang LI
1
;
Jiajin WU
;
Xinhua LU
;
Min LI
;
Muyun WEI
Author Information
1. 上海交通大学医学院附属仁济医院检验科，上海 200127
- Keywords:
Polyomavirus;
Torque teno virus;
Droplet digital PCR
- From:
Chinese Journal of Laboratory Medicine
2024;47(9):1067-1072
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a multiplex detection method for monitoring viruses post-kidney transplantation based on droplet digital PCR (ddPCR) technology, evaluate its detection performance, and discuss its potential clinical application.Methods:We developed a ddPCR assay for the simultaneous detection of polyomavirus type 1 virus (BKV), polyomavirus type 2 virus (JCV), and torque teno virus (TTV), assessing its conformity rate, repeatability, and detection limit. 69 clinical urine samples were collected at the Shanghai Jiao Tong University School of Medicine affiliated with Renji Hospital between June and September 2023. The qPCR and ddPCR methods were employed to analyze the results, respectively. Spearman correlation analysis, Bland-Altman analysis, and Wilcoxon paired rank sum test were applied to analyze the detection results of the two methodologies.Results:The constructed ddPCR method had a 7/7 concordance rate for the seven reference samples. The precision reference samples were tested ten times, respectively, and their coefficients of variation were less than 10%. The detection limit was 10 copies/μl. The concordance rates of ddPCR and qPCR for BKV and JCV detection were more than 95% (66/69, 68/69). The correlation coefficients (R) of BKV and JCV viral loads were 0.874 and 0.840, respectively. The Bland-Altman analysis showed that the mean differences in BKV and JCV viral load detection between the two methods were -0.34 and -0.187, respectively. The Wilcoxon paired rank sum test showed that there were no statistically significant differences in the detection results of polyomaviruses between the two methods ( P>0.05). Conclusion:A ddPCR-based multiplex detection method for viruses post-renal transplantation was successfully developed, confirming its superior attributes of high accuracy, high sensitivity, and multi-detection capability. This method could be used for the quantitative detection of BKV, JCV, and TTV in clinical urine samples.
