Ultrasensitive ELISA Developed for Diagnosis

• DOI: 10.20944/preprints201905.0328.v1
• Publication Date: 2019-05-29T00:14:16Z
• Authors: Kanako Iha; Mikio Inada; Naoki Kawada; Kazunari Nakaishi; Satoshi Watabe; Yong Hong Tan; Chieh Shen; Liang-Yin Ke; Teruki Yoshimura; Etsuro Ito
• Keywords: Absorbance

For the diagnosis of disease, ability to quantitatively detect trace amounts causal proteins from bacteria/viruses as biomarkers in patient specimens is highly desirable. Here we introduce a simple, rapid, and colorimetric assay de novo, ultrasensitive detection method. This consists sandwich enzyme-linked immunosorbent (ELISA) thionicotinamide-adenine dinucleotide (thio-NAD) cycling, forming an ELISA, which signal substrate (i.e., thio-NADH) accumulates triangular manner, accumulated thio-NADH measured at its maximum absorption wavelength 400 nm. We have successfully achieved limit ca. 10–18 moles/assay for target protein. As example infectious disease detection, HIV-1 p24 could be 0.0065 IU/assay 10−18 moles/assay), marker lifestyle-related adiponectin detected 2.3 × 10−19 moles/assay. In particular, despite long-held belief that urine can only using radioisotope, our ELISA was able urinary adiponectin. method versatile, because simply changing antibody enables various proteins. system requires measurement absorbance, thus it equipment easily obtained by medical facilities, facilitates hospitals clinics. Moreover, describe expansion non-amplification nucleic acid acids hybridization. These novo methods will enable accurate diagnosis.