PMID- 20551144
OWN - NLM
STAT- MEDLINE
DCOM- 20101230
LR  - 20211020
IS  - 1531-2267 (Electronic)
IS  - 1094-8341 (Print)
IS  - 1094-8341 (Linking)
VI  - 42A
IP  - 1
DP  - 2010 Sep
TI  - Multiplexed quantitative real-time PCR to detect 22q11.2 deletion in patients 
      with congenital heart disease.
PG  - 52-60
LID - 10.1152/physiolgenomics.00073.2010 [doi]
AB  - 22q11.2 Deletion syndrome (22q11.2 DS) [DiGeorge syndrome type 1 (DGS1)] occurs 
      in  approximately 1:3,000 live births; 75% of children with DGS1 have severe congenital heart 
      disease requiring early intervention. The gold standard for detection of DGS1 is 
      fluorescence in situ hybridization (FISH) with a probe at the TUPLE1 gene. 
      However, FISH is costly and is typically ordered in conjunction with a karyotype 
      analysis that takes several days. Therefore, FISH is underutilized and the 
      diagnosis of 22q11.2 DS is frequently delayed, often resulting in profound 
      clinical consequences. Our goal was to determine whether multiplexed, 
      quantitative real-time PCR (MQPCR) could be used to detect the haploinsufficiency 
      characteristic of 22q11.2 DS. A retrospective blinded study was performed on 382 
      subjects who had undergone congenital heart surgery. MQPCR was performed with a 
      probe localized to the TBX1 gene on human chromosome 22, a gene typically deleted 
      in 22q11.2 DS. Cycle threshold (C(t)) was used to calculate the relative gene 
      copy number (rGCN). Confirmation analysis was performed with the Affymetrix 6.0 
      Genome-Wide SNP Array. With MQPCR, 361 subjects were identified as nondeleted 
      with an rGCN near 1.0 and 21 subjects were identified as deleted with an rGCN 
      near 0.5, indicative of a hemizygous deletion. The sensitivity (21/21) and 
      specificity (361/361) of MQPCR to detect 22q11.2 deletions was 100% at an rGCN 
      value drawn at 0.7. One of 21 subjects with a prior clinical (not genetically 
      confirmed) DGS1 diagnosis was found not to carry the deletion, while another 
      subject, not previously identified as DGS1, was detected as deleted and 
      subsequently confirmed via microarray. The MQPCR assay is a rapid, inexpensive, 
      sensitive, and specific assay that can be used to screen for 22q11.2 deletion 
      syndrome. The assay is readily adaptable to high throughput.
FAU - Tomita-Mitchell, Aoy
AU  - Tomita-Mitchell A
AD  - Division of Cardiovascular Surgery, Department of Surgery, Medical College of 
      Wisconsin, Milwaukee, Wisconsin 53226, USA. amitchell@mcw.edu
FAU - Mahnke, Donna K
AU  - Mahnke DK
FAU - Larson, Joshua M
AU  - Larson JM
FAU - Ghanta, Sujana
AU  - Ghanta S
FAU - Feng, Ying
AU  - Feng Y
FAU - Simpson, Pippa M
AU  - Simpson PM
FAU - Broeckel, Ulrich
AU  - Broeckel U
FAU - Duffy, Kelly
AU  - Duffy K
FAU - Tweddell, James S
AU  - Tweddell JS
FAU - Grossman, William J
AU  - Grossman WJ
FAU - Routes, John M
AU  - Routes JM
FAU - Mitchell, Michael E
AU  - Mitchell ME
LA  - eng
GR  - 1R21-HD-060309-01/HD/NICHD NIH HHS/United States
GR  - R2-HD-060309-02S1/HD/NICHD NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20100615
PL  - United States
TA  - Physiol Genomics
JT  - Physiological genomics
JID - 9815683
SB  - IM
MH  - Chromosomes, Human, Pair 22/*genetics
MH  - DNA Copy Number Variations/genetics
MH  - DiGeorge Syndrome/genetics
MH  - Heart Defects, Congenital/*genetics
MH  - Humans
MH  - Polymerase Chain Reaction/*methods
MH  - Polymorphism, Single Nucleotide/genetics
MH  - Retrospective Studies
PMC - PMC2957771
EDAT- 2010/06/17 06:00
MHDA- 2010/12/31 06:00
PMCR- 2010/06/15
CRDT- 2010/06/17 06:00
PHST- 2010/06/17 06:00 [entrez]
PHST- 2010/06/17 06:00 [pubmed]
PHST- 2010/12/31 06:00 [medline]
PHST- 2010/06/15 00:00 [pmc-release]
AID - physiolgenomics.00073.2010 [pii]
AID - PG-00073-2010 [pii]
AID - 10.1152/physiolgenomics.00073.2010 [doi]
PST - ppublish
SO  - Physiol Genomics. 2010 Sep;42A(1):52-60. doi: 10.1152/physiolgenomics.00073.2010. 
      Epub 2010 Jun 15.