Laser desorption mass spectrometry for point mutation detection

N. I. Taranenko, Karla Matteson, C. N. Chung, Y. F. Zhu, L. Y. Chang, S. L. Allman, L. Haff, S. A. Martin, C. H. Chen

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

A point mutation can be associated with the pathogenesis of inherited or acquired diseases. Laser desorption mass spectrometry coupled with allele specific polymerase chain reaction (PCR) was first used for point mutation detection. G551D is one of several mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene present in 1-3% of the mutant CFTR alleles in most European populations. In this work, two different approaches were pursued to detect G551D point mutation in the cystic fibrosis gene. The strategy is to amplify the desired region of DNA template by PCR rising two primers that overlap one base at the site of the point mutation and width vary in size. If the two primers based on the normal sequence match the target DNA sequence, a normal PCR product will be produced. However, if the alternately sized primers that match the mutant sequence recognize the target DNA, an abnormal PCR product will be produced. Thus, the mass spectrometer can be used to identify patients that are homozygous normal, heterozygous for a mutation or homozygous abnormal at a mutation site. Another approach to identify similar mutations is the use of sequence specific restriction enzymes which respond to changes in the DNA sequence. Mass spectrometry is used to detect the length of the restriction fragments generated by digestion of a PCR generated target fragment.

Original languageEnglish (US)
Pages (from-to)87-94
Number of pages8
JournalGenetic Analysis - Biomolecular Engineering
Volume13
Issue number4
DOIs
StatePublished - Jan 1 1996

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Polymerase chain reaction
Point Mutation
Mass spectrometry
Desorption
Mass Spectrometry
Lasers
Polymerase Chain Reaction
Cystic Fibrosis Transmembrane Conductance Regulator
Mutation
DNA sequences
DNA-Directed DNA Polymerase
Reaction products
DNA
Genes
Alleles
Mass spectrometers
Regulator Genes
Cystic Fibrosis
Digestion
Enzymes

All Science Journal Classification (ASJC) codes

  • Applied Microbiology and Biotechnology
  • Genetics

Cite this

Taranenko, N. I., Matteson, K., Chung, C. N., Zhu, Y. F., Chang, L. Y., Allman, S. L., ... Chen, C. H. (1996). Laser desorption mass spectrometry for point mutation detection. Genetic Analysis - Biomolecular Engineering, 13(4), 87-94. https://doi.org/10.1016/S1050-3862(95)00154-9

Laser desorption mass spectrometry for point mutation detection. / Taranenko, N. I.; Matteson, Karla; Chung, C. N.; Zhu, Y. F.; Chang, L. Y.; Allman, S. L.; Haff, L.; Martin, S. A.; Chen, C. H.

In: Genetic Analysis - Biomolecular Engineering, Vol. 13, No. 4, 01.01.1996, p. 87-94.

Research output: Contribution to journalArticle

Taranenko, NI, Matteson, K, Chung, CN, Zhu, YF, Chang, LY, Allman, SL, Haff, L, Martin, SA & Chen, CH 1996, 'Laser desorption mass spectrometry for point mutation detection', Genetic Analysis - Biomolecular Engineering, vol. 13, no. 4, pp. 87-94. https://doi.org/10.1016/S1050-3862(95)00154-9
Taranenko, N. I. ; Matteson, Karla ; Chung, C. N. ; Zhu, Y. F. ; Chang, L. Y. ; Allman, S. L. ; Haff, L. ; Martin, S. A. ; Chen, C. H. / Laser desorption mass spectrometry for point mutation detection. In: Genetic Analysis - Biomolecular Engineering. 1996 ; Vol. 13, No. 4. pp. 87-94.
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