Detection of point mutations by solid-phase methods

Abstract

Several techniques exist that permit the efficient distinction among characterized DNA sequence variants. In this review we discuss a number of such analytic procedures. These techniques all take advantage of a variety solid supports to prepare and analyze reaction products. The described diagnostic principles are now being applied for the development of miniaturized assay formats, suitable for automated detection of large sets of sequences in clinical samples.

Mutation detection by solid phase primer extension

“…The objective of this study was the development of a mutation detection system based upon primer extension methodologies (Sokolov, 1989;Syvanen et al, 1990Syvanen et al, , 1992Kuppuswamy et al, 1991;Lee and Anvret, 1991;Livak and Hainer, 1994;Syvanen and Landegren, 1994). Oligonucleotide separation is required for specific signal detection from each oligonucleotide that is extended by T7 DNA polymerase.…”

“…Screening methods (Myers et al, 1985;Orita et al, 1989a,b;Sheffield et al , 1989) can detect only the presence of a mutation in a short DNA region, with subsequent definition of the mutation by sequencing. Mutations also can be identified with the use of oligonucleotides as specific probes for hybridization (Wallace et al, 1981;Conner et al, 1983) or enzymatic reactions (Landegren et al, 1988;Gibbs et al, 1989;Sokolov, 1989;Syvanen et al, 1990Syvanen et al, , 1992Barany, 1991;Kuppuswamy et al, 1991;Lee and Anvret, 1991;Livak and Hainer, 1994;Syvanen and Landegren, 1994).…”

A genetic test which can be used to diagnose adult-type hypolactasia in children

“…32 In conclusion, our results highlight the applicability of the genetic test for C/T -13910 variant in the diagnosis of adult-type hypolactasia. This test is easily performed with a semiautomated analysis from a drop of peripheral blood without fasting, 22 making this test suitable not just for diagnostic purposes at the individual level but also as a screening method for population studies. In our study parents had a positive attitude towards genetic testing of adult-type hypolactasia in their children and only a few refused to participate in the study.…”

High Throughput Genotyping Technologies for Pharmacogenomics

“…[26,27] For example, incorporation of a moiety such as biotin to the 5′ end of the query primer allows collection of the single-base extended primer and measurement of the signal produced by the tag. In this method, the wells of the reaction plate are coated with streptavidin, which binds with high affinity to the biotin.…”