Page 58 - Clinical Biochemistry

• Reverse transcribe mRNA (gene expression).

Objectives:

By the end of this chapter, the student should be aware of screening for viral hepatitis,
HCV drug resistance testing, detection of genetic mutations and DNA fingerprinting.

Chapter 6:

Enzyme linked immunosorbent assay.

Outlines:

• Applications of ELISA
• Restriction Fragment Length Polymorphisms
• The Use of Microsatellites in DNA Fingerprinting
• Applications of DNA Fingerprinting
• DNA Microarrays

ELISA
• From the late 1960s, ELISA tests play a role in diagnostic research over 50 years.
• The origin of ELISA was the idea of finding an alternative method to substitute
radioimmunoassay (RIA) in immunoassay.
• Before the invention of ELISA, RIA was the only method to conduct immunoassay.
• The First paper introduced this technique was published in 1960 by Rosalyn Sussman
Yalow and Solomon Berson.
• However, the concerns of potential safety and health problems coming with the initiation
of RIA. Scientists proposed to find another labelling method to replace radioactive label.
• Some of them came up with an idea to use enzyme labels in immunoassay.
• But, many thought it is impossible to link the enzyme, such as large molecule, to an antibody
or antigen without affecting their bioactivity in conjugating reaction.
• In 1966-1970, scientists continually polish and improve methods, including resolving two
critical issues for ELISA, which are “reaction associated color changing” and “removing
non-specific conjugation”.