A microfluidic magnetic bead-based proximity ligation assay device for the quantification of TNF-α in human plasma

The prevalence of autoimmune diseases affect millions of patients worldwide and its incidence has increased over the years. Here we report on the use of a microfluidic device and homemade portable integrated real-time PCR (qPCR) device for the quantification of tumor necrosis factor-α (TNF-α) at the point-of-care (POC). Our device is a disposable cyclic polyolefin (COP)-based microfluidic chip, and the assay is a magnetic bead-based proximity ligation assay. The data presented herein show on-chip TNF-α quantification within the clinically relevant range of 5-100 pg/ml in both buffer and human plasma. Further development could lead to a potential method for monitoring anti-inflammatory therapy.

[1]  Javier Berganzo,et al.  A novel Real Time micro PCR based Point-of-Care device for Salmonella detection in human clinical samples. , 2012, Biosensors & bioelectronics.

[2]  J. Ruano-Lopez,et al.  A Self-contained Diagnostic Platform for DNA Concentration, Elution, and qPCR Inside a LabCard with Stored Reagents☆ , 2012 .

[3]  Mats Lekander,et al.  Levels of Tumour Necrosis Factor-Alpha and Interleukin-6 in Severely Ill Patients with Eating Disorders , 2010, Neuropsychobiology.

[4]  P. Scully,et al.  Plasma Cytokine Profiles in Females With Irritable Bowel Syndrome and Extra-Intestinal Co-Morbidity , 2010, The American Journal of Gastroenterology.

[5]  Nadir Arber,et al.  Inflammation and colorectal cancer. , 2009, Current opinion in pharmacology.

[6]  Jonas Jarvius,et al.  Cytokine detection by antibody-based proximity ligation. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[7]  Antonio López-Vázquez,et al.  High serum tumor necrosis factor-α levels are associated with lack of response to infliximab in fistulizing Crohn's disease , 2002, American Journal of Gastroenterology.

[8]  H. Lorenz,et al.  Perspectives for TNF-α-targeting therapies , 2002, Arthritis research.

[9]  U. Landegren,et al.  Protein detection using proximity-dependent DNA ligation assays , 2002, Nature Biotechnology.

[10]  J. Cavaillon,et al.  Variable estimates of cytokine levels produced by commercial ELISA kits: results using international cytokine standards. , 1995, Journal of immunological methods.