Prediction of acute rejection in kidney transplanted patients based on the point-of-care isothermal molecular diagnostics platform

• Published in: Biosensors & bioelectronics Vol. 199; p. 113877
• Main Authors: Nguyen, Van Dan, Van Nguyen, Hau, Seo, Jung Woo, Lee, Sang Ho, Seo, Tae Seok
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 01.03.2022
• Subjects: Acute rejection (AR); AR prediction Model; Biosensing Techniques; Centrifugal microfluidics; Graft Rejection - diagnosis; Graft Rejection - genetics; Humans; Kidney; Kidney transplantation; Loop mediated isothermal amplification; Nucleic Acid Amplification Techniques; Pathology, Molecular; Point-of-Care Systems; Urinary mRNA; Acute rejection (AR); Urinary mRNA; Centrifugal microfluidics; AR prediction Model; Loop mediated isothermal amplification; Kidney transplantation
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2021.113877

In this study, we proposed an advanced point-of-care molecular diagnostic technology to evaluate the acute rejection (AR) in kidney transplanted patients. On the contrary to the conventional PCR method, we developed a colorimetric loop mediated isothermal amplification (LAMP) for quantitative analysis of the six biomarkers related to AR (CD3ϵ, IP-10, Tim-3-HAVCR2, CXCL9, PSMB9, C1QB) with a reference gene (18S rRNA). Using urinary cDNA samples of transplanted patients, it turned out that three biomarkers among six, namely IP-10, Tim-3-HAVCR2 and C1QB, have significant discrepancy in quantity between the stable graft (STA) patient and the AR patient. The AR prediction model using these three biomarkers was established, which could estimate the immune-rejection in the patients with 93.3% of accuracy. For the point-of-care (POC) molecular diagnostics for the AR evaluation, we constructed a centrifugal microfluidic platform, in which the RNA extraction from the clinical urinary samples, the quantitative reverse-transcription (RT)-LAMP reaction, and the data analysis based on the AR prediction model could be performed in a serial order. Ten blind clinical samples were analyzed on the POC genetic analyzer, showing 100% match with the validated qPCR data. Thus, the proposed advanced molecular diagnostic platform enables us to perform the timely treatment for the transplanted patients who are suffering from the allograft failure and side effects such as infection and malignancy. •A colorimetric RT-LAMP was applied for quantitative analysis of the six biomarkers related to AR.•IP-10, Tim-3-HAVCR2 and C1QB biomarkers have significant quantity discrepancy between STA and AR patients.•An AR prediction model using three biomarkers (93.3% of accuracy) was constructed.•A POC centrifugal microfluidic platform for biomarker quantification was proposed.•Ten blind clinical samples were successfully analyzed on the POC genetic analyzer.