Immunoglobulin E Detection Method Based on Cascade Enzymatic Reaction Utilizing Portable Personal Glucose Meter

Abstract

We herein describe a cascade enzymatic reaction (CER)-based IgE detection method utilizing a personal glucose meter (PGM), which relies on alkaline phosphatase (ALP) activity that regulates the amount of adenosine triphosphate (ATP). The amount of sandwich assay complex is determined according to the presence or absence of the target IgE. Additionally, the ALP in the sandwich assay catalyzes the dephosphorylation of ATP, a substrate of CER, which results in the changes in glucose level. By employing this principle, IgE was reliably detected at a concentration as low as ca. 29.6 ng/mL with high specificity toward various proteins. Importantly, the limit of detection (LOD) of this portable PGM-based approach was comparable to currently commercialized ELISA kit without expensive and bulky analysis equipment as well as complexed washing step. Finally, the diagnostic capability of this method was also successfully verified by reliably detecting IgE present in a real human serum sample with an excellent recovery ratio within 100 ± 6%.

A dual-mode strategy based on β-galactosidase and target-induced DNA polymerase protection for transcription factor detection using colorimetry and a glucose meter

“…[25][26][27] As the signal transduction involved transducing the non-glucose substrates into glucose, only several enzymes, including α-glucosidase, invertase, alkaline phosphatase (ALP), hexokinase, and glucose oxidase, have been chosen to turn the substrates into glucose. [28][29][30][31] Furthermore, the PGM-based signal can be influenced by test conditions, detection surroundings, and device efficiency. Thus, the integration of PGM-based analysis and instrument-based methods can provide more accurate and reliable detection, and more flexible choices than single-readout methods.…”

Washing-Free and Label-Free Onsite Assay for Inorganic Pyrophosphatase Activity Using a Personal Glucose Meter

“…In recent years, a personal glucose meter (PGM), which is a self-monitoring kit that monitors the glucose level in the blood, has been used as a biosensing platform to detect non-glucose target analytes which regulate glucose concentration. − As pioneering research, Xiang and Lu developed PGM-based biosensing systems to detect various biomolecular targets, such as nucleic acids, proteins, and small molecules, by employing an invertase-conjugated DNA probe . In these systems, target analytes bound to the specific aptamer probe resulted in the release of an invertase-conjugated DNA probe which then catalyzed the hydrolysis of sucrose to glucose.…”