Early diagnosis with ultrasensitive ELISA

Adv Clin Chem. 2021:101:121-133. doi: 10.1016/bs.acc.2020.06.002. Epub 2020 Jul 7.


Accurate, rapid and simple detection methods are required to facilitate early diagnosis of various disorders including infectious and lifestyle diseases as well as cancer. These detection approaches reduce the window of infection, i.e., the period between infection and reliable detection. Optimally, these methods should target protein as an indicator of pathogenic microbes as well as other biomarkers. For example, although nucleic acid is easily detected by polymerase chain reaction (PCR), these markers are also present in dead microbes, and, in the case of mRNA, it is not known whether this target was successfully translated. Accordingly, early diagnostic approaches require the development of ultrasensitive protein detection methods. In this chapter, we introduce an ultrasensitive enzyme-linked immunosorbent assay (ELISA) which combines a traditional sandwich-based immunoassay with thionicotinamide adenine dinucleotide (thio-NAD) cycling. The performance characteristics of this unique approach are reviewed as well as its potential role in providing a novel and ultrasensitive diagnostic tool in the clinical laboratory.

Keywords: Cancer marker; Infectious disease; Lifestyle disease; Non-amplification nucleic acid detection; Single-cell detection; Ultrasensitive ELISA; Window period.

Publication types

  • Review

MeSH terms

  • Biomarkers
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Humans
  • Limit of Detection
  • Nucleic Acids / chemistry
  • Nucleic Acids / metabolism*
  • Proteins / chemistry
  • Proteins / metabolism*


  • Biomarkers
  • Nucleic Acids
  • Proteins