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

Forskningsoutput: TidskriftsbidragArtikel i vetenskaplig tidskriftPeer review

Sammanfattning

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.
Originalspråkengelska
Sidor (från-till)1427-1436
TidskriftJournal of Assisted Reproduction and Genetics
Volym31
Nummer11
DOI
StatusPublished - 2014

Ämnesklassifikation (UKÄ)

  • Farmakologi och toxikologi
  • Läkemedelskemi

Fingeravtryck

Utforska forskningsämnen för ”Genotyping common FSHR polymorphisms based on competitive amplification of differentially melting amplicons (CADMA).”. Tillsammans bildar de ett unikt fingeravtryck.

Citera det här