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

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

Research output: Contribution to journalArticlepeer-review

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.
Original languageEnglish
Pages (from-to)1427-1436
JournalJournal of Assisted Reproduction and Genetics
Volume31
Issue number11
DOIs
Publication statusPublished - 2014

Subject classification (UKÄ)

  • Pharmacology and Toxicology
  • Medicinal Chemistry

Free keywords

  • CADMA
  • Fertility treatment
  • FSHR polymorphisms
  • Genotype analysis
  • Pharmacogenetics

Fingerprint

Dive into the research topics of 'Genotyping common FSHR polymorphisms based on competitive amplification of differentially melting amplicons (CADMA).'. Together they form a unique fingerprint.

Cite this