Table 1.
Clinical characteristics of kidney allograft recipients.
Fig 1.
Workflow of patients, urine samples, and experimental design.
For validation of urinary cell mRNA, 90 urine samples were collected from 88 kidney transplant patients. Two PCR platforms were used to process the 90 samples, and 79 of the samples passed the quality control (QC) thresholds of TGF-β1 and 18S rRNA copies/μg total RNA; 11 did not pass. Data were statistically analyzed with the QC-passed samples.
Fig 2.
The mRNA levels of the CTOT4 formula and CD3ε, IP-10, and 18S rRNA in qPCR.
(A) The mRNA levels of the CTOT4 formula, (C) CD3ε, (E) IP-10, and (G) 18S rRNA between two groups were analyzed by qPCR, respectively. (B, D, F, H) The results corresponding ROC curve analyses for the CTOT4 formula and three genes, respectively.
Fig 3.
The mRNA levels of the CTOT4 formula and the mRNAs in ddPCR.
(A) The CTOT4 formula, (C) CD3ε, (E) IP-10, and (G) 18S rRNA represented the absolute mRNA levels between two groups analyzed by ddPCR, respectively. (B, D, F, H) The results corresponding ROC curve analyses for the CTOT4 formula and three genes, respectively.
Fig 4.
Comparison of several methods for mining data and two PCR platforms.
(A-B) The area under the curve (AUC) values for urinary CD3ε and IP-10 mRNA levels normalized by the total RNA amount (absolute), absolute copy number of 18S rRNA, or the 2-ΔΔCт method in qPCR platform. (C) ROC curves for two-gene set (CD3ε and IP-10) normalized by absolute copy number of 18S rRNA in qPCR and ddPCR platforms.