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Journal of Assisted Reproduction and Genetics

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01-11-2014 | Technological Innovations

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

Authors: Tanni Borgbo, Lasse Sommer Kristensen, Ida Lindgren, Claus Yding Andersen, Lise Lotte Hansen

Published in: Journal of Assisted Reproduction and Genetics | Issue 11/2014

Abstract

Purpose

To provide an improved platform for simple, reliable, and cost-effective genotyping.

Background

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.

Methods

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.

Results

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.

Conclusions

The new assays provide a reliable, fast and user-friendly genotyping method facilitating a wider implication in clinical practise.

Available only for authorised users

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Title: Genotyping common FSHR polymorphisms based on competitive amplification of differentially melting amplicons (CADMA).
Authors: Tanni Borgbo
Lasse Sommer Kristensen
Ida Lindgren
Claus Yding Andersen
Lise Lotte Hansen
Publication date: 01-11-2014
Publisher: Springer US
Published in: Journal of Assisted Reproduction and Genetics / Issue 11/2014
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI: https://doi.org/10.1007/s10815-014-0329-6

Keynote webinar | Spotlight on medication adherence

Springer Medicine

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

Abstract

Purpose

Background

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Background

Methods

Results

Conclusions

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