PMID- 10024431
OWN - NLM
STAT- MEDLINE
DCOM- 19990415
LR  - 20091119
IS  - 0890-8508 (Print)
IS  - 0890-8508 (Linking)
VI  - 13
IP  - 1
DP  - 1999 Feb
TI  - Screening for hereditary fructose intolerance mutations by reverse dot-blot.
PG  - 35-40
AB  - An assay is described which is useful for genetic screening of the two most
      prevalent mutations that cause hereditary fructose intolerance (HFI). Both
      mutations lie within exon 5 of the aldolase B gene. Amplification of exon 5 from 
      genomic DNA isolated from peripheral lymphocytes using biotinylated aldolase
      B-specific primers yields a biotin-tagged probe. This probe is hybridized to
      complementary poly(dT)-tailed allele specific oligonucleotides (ASOs) that are
      bound to a nylon membrane. The length of the ASOs, the amount bound to the
      membrane and the time of hybridization are optimized for discrimination of all
      four alleles under the same hybridization conditions. Detection of biotinylated
      amplified DNA is performed by creating an avidin-alkaline phosphatase complex and
      visualization by chemiluminescence. This assay can rapidly detect the two
      mutations, A149P and A174D, which cause >70% of HFI worldwide, and offers a rapid
      and sensitive assay that is much less invasive for the diagnosis of this often
      difficult to diagnose disorder.
CI  - Copyright 1999 Academic Press.
FAU - Lau, J
AU  - Lau J
AD  - Department of Biology, Boston University, MA 02215, USA.
FAU - Tolan, D R
AU  - Tolan DR
LA  - eng
GR  - PHS-DK54321/DK/NIDDK NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Mol Cell Probes
JT  - Molecular and cellular probes
JID - 8709751
RN  - 0 (Isoenzymes)
RN  - 0 (Oligonucleotide Probes)
RN  - EC 4.1.2.13 (Fructose-Bisphosphate Aldolase)
SB  - IM
MH  - Alleles
MH  - Amino Acid Substitution
MH  - Biotinylation
MH  - DNA Mutational Analysis/*methods
MH  - Fructose Intolerance/diagnosis/*genetics
MH  - Fructose-Bisphosphate Aldolase/*genetics
MH  - Gene Frequency
MH  - Genetic Testing/*methods
MH  - Genotype
MH  - Humans
MH  - Isoenzymes/*genetics
MH  - Nucleic Acid Hybridization/*methods
MH  - Oligonucleotide Probes
MH  - *Point Mutation
MH  - Polymerase Chain Reaction
EDAT- 1999/02/20 00:00
MHDA- 1999/02/20 00:01
CRDT- 1999/02/20 00:00
PHST- 1999/02/20 00:00 [pubmed]
PHST- 1999/02/20 00:01 [medline]
PHST- 1999/02/20 00:00 [entrez]
AID - S0890-8508(98)90208-0 [pii]
AID - 10.1006/mcpr.1998.0208 [doi]
PST - ppublish
SO  - Mol Cell Probes. 1999 Feb;13(1):35-40. doi: 10.1006/mcpr.1998.0208.