Microfluidic LIPS for serum antibody detection: demonstration of a rapid test for HSV-2 infection

Abstract

There is great interest in point-of-care antibody testing for the diagnosis of infectious and autoimmune diseases. As a first step in the development of self-contained and miniaturized devices for highly quantitative antibody detection, we demonstrate the application of Luciferase Immunoprecipitation Systems (LIPS) technology in a microfluidic format. Protein A/G was immobilized on the walls of PDMS-glass microchannels of 500 nL volume. The assay proceeds with the simultaneous introduction of plasma and Renilla luciferase–tagged antigens. Following washing, coelenterazine substrate was added and bound antigen-luciferase measured by chemiluminescence. Total assay time, including rinsing and detection, is under ten minutes. Using these stable microfluidic devices, high diagnostic performance (100% sensitivity and 100% specificity) was achieved for the diagnosis of HSV-2 infection. Based on these findings, the LIPS microfluidic format should readily lend itself to automation and the transfer to portable instrumentation.

Point of care diagnostics for sexually transmitted infections: perspectives and advances

“…Recently, promising microfluidic technology utilizing luciferase coupled antigens for detection of HSV-2 antibodies was demonstrated [101]. The luciferase immunoprecipitation system presented in the publication relies on light emitting recombinant antigens to measure antibody titers, and demonstrated 100% sensitivity and specificity for detection of HSV-2 antibodies in 20 human plasma samples [101]. With a detection time of <10 min, this platform has great potential for automation and portability, although further evaluation will be required.…”

Anti-Ro and anti-La autoantibody profiling in Norwegian patients with primary Sjögren’s syndrome using luciferase immunoprecipitation systems (LIPS)

“…In conclusion, our results show that LIPS could substitute ELISA for the diagnosis of pSS in the future, especially if the technology was available as a microfluidics device for point-of-care medicine (10). Nevertheless, further studies using a larger control cohort are required to support the claim for the high specificity of the assay.…”

Lab-on-a-Chip and Fluid Manipulation Applications

“…For example, soda lime glass is one of the cheapest and most commonly used forms of glass, but can contain a large amount of contaminants such as aluminum. Other target molecules include microRNAs [95,99], a biomarker for cancer, DNA [88,93,98,103,107,127], herpes simplex-2 virus infection [78], and common drinking water contaminates including E. coli [118], pesticides [95,108], synthetic estrogen in river water [100], and heavy metals [76, 134À136]. The best optical properties would be achieved in either fused-silica or quartz but these types of materials are significantly more expensive.…”

“…The device uses an array of microelectrodes as a transducer and ferro/ferricyanide entrapping liposomes as signal reporters to quantify the concentration of DNA in the range of 1À50 fM in 6 min using only a simple potentiostat [133]. This could potentially lead to a single device that can be used to detect several different target molecules at once [78]. The device is fully portable and relatively inexpensive (,$50) [93], when compared to some other microfluidics devices that can easily cost well over a few hundred dollars.…”