PMID- 11551105
OWN - NLM
STAT- MEDLINE
DCOM- 20020304
LR  - 20061115
IS  - 1090-6576 (Print)
IS  - 1090-6576 (Linking)
VI  - 5
IP  - 2
DP  - 2001 Summer
TI  - PCR-based methylation testing for Prader-Willi or Angelman syndromes using 
      archived fixed-cell suspensions.
PG  - 153-5
AB  - All Prader-Willi syndrome (PWS) and 75% of Angelman syndrome (AS) patients have 
      specific DNA methylation pattern alterations that can be used for diagnostic 
      evaluation. The methylation testing identifies a significantly higher proportion 
      of patients as compared to fluorescence in situ hybridization (FISH)-based 
      microdeletion analysis and is thus a useful diagnostic evaluation for clinically 
      suspect, but FISH-negative, patients. We used two independent PCR-based protocols 
      for methylation testing on fixed cell specimens archived after FISH analyses. 
      Changes in DNA methylation due to the procedure of cell fixation were ruled out 
      by testing control specimens before and after fixation. Then methylation testing 
      was carried out on 20 standard fixed-cell supsensions from people suspected for 
      PWS or AS. These fixed specimens were stored after negative FISH analysis for up 
      to 4 years at 4 degrees C in 3:1 methanol/acetic acid. Methylation patterns 
      associated with AS (one specimen) and with PWS (one specimen) were identified for 
      both protocols. The observed methylation patterns were concordant with the 
      phenotypes of the positive individuals and for the two protocols used. We have, 
      thus, shown that archived fixed-cell suspensions from individuals suspected as 
      PWS or AS that were negative for cytogenetic/FISH microdeletions, can now be 
      re-evaluated with PCR-based methylation testing without the need for additional 
      blood samples from the previously studied individuals.
FAU - Velinov, M
AU  - Velinov M
AD  - New York State Institute for Basic Research in Developmental Disabilities, Staten 
      Island 10314, USA. milen.velinov@omr.state.ny.us
FAU - Gu, H
AU  - Gu H
FAU - Shah, K
AU  - Shah K
FAU - Genovese, M
AU  - Genovese M
FAU - Duncan, C
AU  - Duncan C
FAU - Kupchik, G
AU  - Kupchik G
FAU - Jenkins, E C
AU  - Jenkins EC
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Genet Test
JT  - Genetic testing
JID - 9802546
RN  - 0 (Suspensions)
SB  - IM
MH  - Angelman Syndrome/diagnosis/*genetics
MH  - Chromosomes, Human, Pair 15/*genetics
MH  - CpG Islands
MH  - *DNA Methylation
MH  - Genomic Imprinting
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - *Polymerase Chain Reaction
MH  - Prader-Willi Syndrome/diagnosis/*genetics
MH  - Preservation, Biological
MH  - Specimen Handling/*methods
MH  - Suspensions
MH  - Time Factors
MH  - Tissue Fixation
EDAT- 2001/09/12 10:00
MHDA- 2002/03/05 10:01
CRDT- 2001/09/12 10:00
PHST- 2001/09/12 10:00 [pubmed]
PHST- 2002/03/05 10:01 [medline]
PHST- 2001/09/12 10:00 [entrez]
AID - 10.1089/109065701753145655 [doi]
PST - ppublish
SO  - Genet Test. 2001 Summer;5(2):153-5. doi: 10.1089/109065701753145655.