Highly Sensitive Lab on a Chip (LOC) Immunoassay for Early Diagnosis of Respiratory Disease Caused by Respirable Crystalline Silica (RCS).

Abstract

Respirable crystalline silica (RCS) produced in mining and construction industries can cause life-threatening diseases such as silicosis, lung cancer, and chronic obstructive pulmonary disease (COPD). These diseases could be more effectively treated and prevented if RCS-related biomarkers were identified and measured at an early stage of disease progression, which makes development of a point of care test (POCT) platform extremely desirable for early diagnosis. In this work, a new, highly sensitive lab on a chip (LOC) immunoassay has been designed, developed, and characterized for tumor necrosis factor α (TNF-α), a protein biomarker that causes lung inflammation due to RCS exposure. The designed LOC device is composed of four reservoirs for sample, enzyme conjugated detection antibody, wash buffer, and chemiluminescence substrate in liquid form, along with three spiral reaction chambers for test, positive control, and negative control. All reservoirs and spiral microchannels were connected in series and designed to perform sequential delivery of immunoassay reagents with minimal user intervention. The developed LOC measured TNF-α concentrations as low as 16 pg/mL in plasma from RCS-exposed rats and also had a limit of detection (LOD) of 0.5 pg/mL in spiked artificial serum. In addition, the analysis time was drastically reduced to about 30 min, as opposed to hours in conventional methods. Successful implementation of a highly sensitive, chemiluminescence-based immunoassay on a preloaded LOC with proper quality control, as reported in this work, can pave the way toward developing a new rapid POCT platform for in-field clinical diagnosis.

• Authors: V Thiyagarajan Upaassana, Sthitodhi Ghosh, Atreyee Chakraborty, M Eileen Birch, Pius Joseph, Jungyoup Han, Bon Ki Ku, Chong H Ahn
• Journal: Analytical chemistry (Anal Chem) Vol. 91 Issue 10 Pg. 6652-6660 (05 21 2019) ISSN: 1520-6882 [Electronic] United States
• PMID: 31012299 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Antibodies, Immobilized
• Biomarkers
• Luminescent Agents
• Tumor Necrosis Factor-alpha
• Silicon Dioxide
• Horseradish Peroxidase

Topics

• Animals
• Antibodies, Immobilized (immunology)
• Biomarkers (blood)
• Enzyme-Linked Immunosorbent Assay (methods)
• Horseradish Peroxidase (chemistry)
• Lab-On-A-Chip Devices
• Limit of Detection
• Luminescent Agents (chemistry)
• Luminescent Measurements
• Male
• Microfluidic Analytical Techniques (instrumentation, methods)
• Point-of-Care Testing
• Rats, Inbred F344
• Silicon Dioxide (toxicity)
• Silicosis (blood, diagnosis)
• Tumor Necrosis Factor-alpha (blood, immunology)