PMID- 18487285
OWN - NLM
STAT- MEDLINE
DCOM- 20080808
LR  - 20190722
IS  - 0009-9147 (Print)
IS  - 0009-9147 (Linking)
VI  - 54
IP  - 7
DP  - 2008 Jul
TI  - Detection of APC gene deletions using quantitative multiplex PCR of short 
      fluorescent fragments.
PG  - 1132-40
LID - 10.1373/clinchem.2007.101006 [doi]
AB  - BACKGROUND: approximately 20% of classic familial adenomatous polyposis (FAP) 
      cases and 70% to 80% of attenuated FAP (AFAP) cases are negative for the 
      APC/MUTYH point mutation. Quantitative multiplex PCR of short fluorescent 
      fragments (QMPSF), a technique for detecting copy number alterations, has been 
      successfully applied to several cancer syndrome genes. We used QMPSF for the APC 
      gene to screen FAP APC/MUTYH mutation-negative families to improve their 
      diagnostic surveillance. METHODS: we set up and validated APC-gene QMPSF using 23 
      negative and 1 positive control and examined 45 (13 FAP and 32 AFAP) unrelated 
      members of APC/MUTYH mutation-negative families for copy number alterations. We 
      confirmed the results using multiplex ligation-dependent probe amplification 
      (MLPA). We used different approaches such as sequencing, quantitative real 
      time-PCR (QRT-PCR), and fluorescence in situ hybridization (FISH) to further 
      characterize the identified deletions. RESULTS: APC QMPSF was capable of 
      detecting deletions with an acceptable variability, as shown by mean values (SD) 
      of allele dosage for the deleted control obtained from intra- and 
      interexperimental replicates [0.52 (0.05) and 0.45 (0.10)]. We detected 3 gross 
      deletions in 13 (23%) of the classic FAP cases analyzed (1 complete gene deletion 
      and 2 partial deletions encompassing exons 9 and 10 and exons 11-15, 
      respectively). No rearrangements were detected in the 32 AFAP cases. CONCLUSIONS: 
      QMPSF is able to detect rearrangements of the APC gene. Our findings highlight 
      the importance of using a copy number alteration methodology as a first step in 
      the routine genetic testing of FAP families in the clinical setting.
FAU - Castellsague, Ester
AU  - Castellsague E
AD  - Translational Research Laboratory, IDIBELL-Institut Catala d'Oncologia, 
      Barcelona, Spain.
FAU - Gonzalez, Sara
AU  - Gonzalez S
FAU - Nadal, Marga
AU  - Nadal M
FAU - Campos, Olga
AU  - Campos O
FAU - Guino, Elisabet
AU  - Guino E
FAU - Urioste, Miguel
AU  - Urioste M
FAU - Blanco, Ignacio
AU  - Blanco I
FAU - Frebourg, Thierry
AU  - Frebourg T
FAU - Capella, Gabriel
AU  - Capella G
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20080516
PL  - England
TA  - Clin Chem
JT  - Clinical chemistry
JID - 9421549
RN  - 0 (Adenomatous Polyposis Coli Protein)
RN  - 0 (Fluorescent Dyes)
RN  - EC 3.2.2.- (DNA Glycosylases)
RN  - EC 3.2.2.- (mutY adenine glycosylase)
SB  - IM
MH  - Adenomatous Polyposis Coli/genetics
MH  - Adenomatous Polyposis Coli Protein/*genetics
MH  - Base Sequence
MH  - DNA Glycosylases/genetics
MH  - Exons
MH  - *Fluorescent Dyes
MH  - *Gene Deletion
MH  - Gene Dosage
MH  - Genes, APC
MH  - Heterozygote
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Molecular Sequence Data
MH  - Point Mutation
MH  - Polymerase Chain Reaction/methods
EDAT- 2008/05/20 09:00
MHDA- 2008/08/09 09:00
CRDT- 2008/05/20 09:00
PHST- 2008/05/20 09:00 [pubmed]
PHST- 2008/08/09 09:00 [medline]
PHST- 2008/05/20 09:00 [entrez]
AID - clinchem.2007.101006 [pii]
AID - 10.1373/clinchem.2007.101006 [doi]
PST - ppublish
SO  - Clin Chem. 2008 Jul;54(7):1132-40. doi: 10.1373/clinchem.2007.101006. Epub 2008 
      May 16.