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Genotyping common FSHR polymorphisms based on competitive amplification of differentially melting amplicons (CADMA).

Borgbo, Tanni; Kristensen, Lasse Sommer; Lindgren, Ida LU ; Andersen, Claus Yding and Hansen, Lise Lotte (2014) In Journal of Assisted Reproduction and Genetics 31(11). p.1427-1436
Abstract
To provide an improved platform for simple, reliable, and cost-effective genotyping. Modern fertility treatments are becoming increasingly individualized in an attempt to optimise the follicular response and reproductive outcome, following controlled ovarian stimulation. As the field of pharmacogenetics evolve, genetic biomarkers such as polymorphisms of the follicle stimulating hormone receptor (FSHR) may be included as a predictive tool for individualized fertility treatment. However, the currently available genotyping methods are expensive, time-consuming or have a limited analytical sensitivity. Here, we present a novel version of "competitive amplification of differentially melting amplicons" (CADMA), providing an improved platform... (More)
To provide an improved platform for simple, reliable, and cost-effective genotyping. Modern fertility treatments are becoming increasingly individualized in an attempt to optimise the follicular response and reproductive outcome, following controlled ovarian stimulation. As the field of pharmacogenetics evolve, genetic biomarkers such as polymorphisms of the follicle stimulating hormone receptor (FSHR) may be included as a predictive tool for individualized fertility treatment. However, the currently available genotyping methods are expensive, time-consuming or have a limited analytical sensitivity. Here, we present a novel version of "competitive amplification of differentially melting amplicons" (CADMA), providing an improved platform for simple, reliable, and cost-effective genotyping. Two CADMA based assays were designed for the two common polymorphisms of the FSHR gene: rs6165 (c.919A > G, p. Thr307Ala, FSHR 307) and rs6166 (c.2039A > G, p. Asn680Ser, FSHR 680). To evaluate the reliability of the new CADMA-based assays, the genotyping results were compared with two conventional PCR based genotyping methods; allele-specific PCR (AS-PCR) and Sanger sequencing. The genotype frequencies for both polymorphisms were 35 % (TT), 42 % (CT), and 23 % (CC), respectively. A 100 % accordance was observed between the CADMA-based genotyping results and sequencing results, whereas 5 discrepancies were observed between the AS-PCR results and the CADMA-based genotyping results. Comparing the CADMA-based assays to (AS-PCR) and Sanger sequencing, the CADMA based assays showed an improved analytical sensitivity and a wider applicability. The new assays provide a reliable, fast and user-friendly genotyping method facilitating a wider implication in clinical practise. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
CADMA, Fertility treatment, FSHR polymorphisms, Genotype analysis, Pharmacogenetics
in
Journal of Assisted Reproduction and Genetics
volume
31
issue
11
pages
1427 - 1436
publisher
Kluwer
external identifiers
  • wos:000345406000005
  • scopus:84912030790
ISSN
1058-0468
DOI
10.1007/s10815-014-0329-6
language
English
LU publication?
yes
id
67217440-4813-486b-85a1-8fd48928ceb7 (old id 4979622)
date added to LUP
2015-02-03 07:09:04
date last changed
2017-11-12 03:43:32
@article{67217440-4813-486b-85a1-8fd48928ceb7,
  abstract     = {To provide an improved platform for simple, reliable, and cost-effective genotyping. Modern fertility treatments are becoming increasingly individualized in an attempt to optimise the follicular response and reproductive outcome, following controlled ovarian stimulation. As the field of pharmacogenetics evolve, genetic biomarkers such as polymorphisms of the follicle stimulating hormone receptor (FSHR) may be included as a predictive tool for individualized fertility treatment. However, the currently available genotyping methods are expensive, time-consuming or have a limited analytical sensitivity. Here, we present a novel version of "competitive amplification of differentially melting amplicons" (CADMA), providing an improved platform for simple, reliable, and cost-effective genotyping. Two CADMA based assays were designed for the two common polymorphisms of the FSHR gene: rs6165 (c.919A > G, p. Thr307Ala, FSHR 307) and rs6166 (c.2039A > G, p. Asn680Ser, FSHR 680). To evaluate the reliability of the new CADMA-based assays, the genotyping results were compared with two conventional PCR based genotyping methods; allele-specific PCR (AS-PCR) and Sanger sequencing. The genotype frequencies for both polymorphisms were 35 % (TT), 42 % (CT), and 23 % (CC), respectively. A 100 % accordance was observed between the CADMA-based genotyping results and sequencing results, whereas 5 discrepancies were observed between the AS-PCR results and the CADMA-based genotyping results. Comparing the CADMA-based assays to (AS-PCR) and Sanger sequencing, the CADMA based assays showed an improved analytical sensitivity and a wider applicability. The new assays provide a reliable, fast and user-friendly genotyping method facilitating a wider implication in clinical practise.},
  author       = {Borgbo, Tanni and Kristensen, Lasse Sommer and Lindgren, Ida and Andersen, Claus Yding and Hansen, Lise Lotte},
  issn         = {1058-0468},
  keyword      = {CADMA,Fertility treatment,FSHR polymorphisms,Genotype analysis,Pharmacogenetics},
  language     = {eng},
  number       = {11},
  pages        = {1427--1436},
  publisher    = {Kluwer},
  series       = {Journal of Assisted Reproduction and Genetics},
  title        = {Genotyping common FSHR polymorphisms based on competitive amplification of differentially melting amplicons (CADMA).},
  url          = {http://dx.doi.org/10.1007/s10815-014-0329-6},
  volume       = {31},
  year         = {2014},
}