PMID- 22192860
OWN - NLM
STAT- MEDLINE
DCOM- 20120517
LR  - 20120130
IS  - 1873-3492 (Electronic)
IS  - 0009-8981 (Linking)
VI  - 413
IP  - 5-6
DP  - 2012 Mar 22
TI  - Identification of rare and frequent variants of the CASR gene by high-resolution 
      melting.
PG  - 605-11
LID - 10.1016/j.cca.2011.12.004 [doi]
AB  - BACKGROUND: Calcium metabolic disorders like familial hypocalciuric hypercalcemia 
      (FHH) and autosomal dominant familial isolated hypoparathyroidism (FIH) can be 
      caused by rare variants of the calcium sensing receptor gene (CASR). Molecular 
      genetic screening of the CASR is often based on DNA sequencing. METHODS: We 
      sought to develop a pre-screening method in the diagnostic procedure and pursued 
      variant scanning by high-resolution melting analysis (HRM) on a LightScanner 
      instrument. We used 50 samples, representing 45 different rare variants, to 
      validate the HRM method. In addition, we implemented small amplicon genotyping of 
      three frequent CASR variants (c.1732+16T/C, c.2956G>T and c.2968A>G). RESULTS: 
      Using HRM, we identified 43 of 45 variants confidently (~96%) while two variants 
      escaped immediate detection. Implementing this method in clinical use further 
      resulted in the identification of seven new CASR variants and nine recurrent. HRM 
      variant scanning, in combination with small amplicon genotyping, provides a 
      simple workflow with reduced sequencing burden. Bioinformatics analyses using two 
      freely available prediction tools (PolyPhen2 and SIFT) for evaluating amino acid 
      substitutions were compared and indicated discrepancies in the prediction for 25% 
      of the variants. CONCLUSION: This study demonstrates the utility of HRM as a 
      pre-screening method, adds 24 novel rare CASR variants, and further emphasizes 
      the importance of clinical decision making based on all available information 
      rather than bioinformatics alone.
CI  - Copyright (c) 2011 Elsevier B.V. All rights reserved.
FAU - Nissen, Peter H
AU  - Nissen PH
AD  - Department of Clinical Biochemistry, Aarhus University Hospital, Tage-Hansens 
      Gade, Denmark. peter.h.nissen@aarhus.rm.dk
FAU - Christensen, Signe E
AU  - Christensen SE
FAU - Ladefoged, Soren A
AU  - Ladefoged SA
FAU - Brixen, Kim
AU  - Brixen K
FAU - Heickendorff, Lene
AU  - Heickendorff L
FAU - Mosekilde, Leif
AU  - Mosekilde L
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20111213
PL  - Netherlands
TA  - Clin Chim Acta
JT  - Clinica chimica acta; international journal of clinical chemistry
JID - 1302422
RN  - 0 (CASR protein, human)
RN  - 0 (Receptors, Calcium-Sensing)
SB  - IM
MH  - Computational Biology
MH  - *Genetic Testing
MH  - Genetic Variation/*genetics
MH  - Humans
MH  - Receptors, Calcium-Sensing/*genetics
MH  - *Transition Temperature
EDAT- 2011/12/24 06:00
MHDA- 2012/05/18 06:00
CRDT- 2011/12/24 06:00
PHST- 2011/11/08 00:00 [received]
PHST- 2011/12/02 00:00 [revised]
PHST- 2011/12/05 00:00 [accepted]
PHST- 2011/12/24 06:00 [entrez]
PHST- 2011/12/24 06:00 [pubmed]
PHST- 2012/05/18 06:00 [medline]
AID - S0009-8981(11)00668-1 [pii]
AID - 10.1016/j.cca.2011.12.004 [doi]
PST - ppublish
SO  - Clin Chim Acta. 2012 Mar 22;413(5-6):605-11. doi: 10.1016/j.cca.2011.12.004. Epub 
      2011 Dec 13.