Accuracy of preimplantation genetic diagnosis in equine in vivo-recovered and in vitro-produced blastocysts

Y. H. Choi, Cecilia Penedo, P. Daftari, I. C. Velez, K. Hinrichs

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Preimplantation genetic diagnosis has great potential in the horse, but information on evaluation of equine embryo biopsy samples is limited. Blastocysts were biopsied using a Piezo drill and methods for whole-genome amplification (WGA) investigated. Results for 33 genetic loci were then compared between biopsy samples from in vitro-produced (IVP) and in vivo-recovered (VIV) blastocysts. Under the experimental conditions described, WGA using the Qiagen Repli-g Midi kit was more accurate than that using the Illustra Genomiphi V2 kit (98.2% vs 25.8%, respectively). Using WGA with the Qiagen kit, three biopsy samples were evaluated from each of eight IVP and 19 VIV blastocysts, some produced using semen from stallions carrying the genetic mutations associated with the diseases hereditary equine regional dermal asthenia (HERDA), hyperkalemic periodic paralysis (HYPP) or polysaccharide storage myopathy 1 (PSSM1). Three of 81 biopsy samples (3.7%) returned <50% accuracy. In the remaining 78 samples, overall accuracy was 99.3% (2556/2574 loci interrogated). Accuracy did not differ significantly between samples from IVP and VIV blastocysts. Allele drop-out in heterozygous loci was 1.6% (17/1035). Accuracy for sex determination was 100%; accuracy for heterozygosity for disease-causing mutations was 97.7% (43/44). In conclusion, Piezo-driven embryo biopsy with WGA has >95% overall accuracy in IVP and VIV embryos, and this technique is suitable for use in a clinical setting.

Original languageEnglish (US)
Pages (from-to)1382-1389
Number of pages8
JournalReproduction, Fertility and Development
Volume28
Issue number9
DOIs
StatePublished - Jan 1 2016

Fingerprint

Preimplantation Diagnosis
Blastocyst
blastocyst
Horses
biopsy
Biopsy
horses
Genome
genome
embryo (animal)
Hyperkalemic Periodic Paralysis
Embryonic Structures
Horse Diseases
Asthenia
sampling
Mandrillus
Inborn Genetic Diseases
Genetic Loci
muscular diseases
genetic disorders

Keywords

  • embryo biopsy
  • genetic screening
  • HERDA
  • HYPP
  • polymerase chain reaction (PCR)
  • PSSM
  • whole genome amplification

ASJC Scopus subject areas

  • Biotechnology
  • Reproductive Medicine
  • Animal Science and Zoology
  • Molecular Biology
  • Genetics
  • Endocrinology
  • Developmental Biology

Cite this

Accuracy of preimplantation genetic diagnosis in equine in vivo-recovered and in vitro-produced blastocysts. / Choi, Y. H.; Penedo, Cecilia; Daftari, P.; Velez, I. C.; Hinrichs, K.

In: Reproduction, Fertility and Development, Vol. 28, No. 9, 01.01.2016, p. 1382-1389.

Research output: Contribution to journalArticle

@article{5c25e3bf7e9b46a8b478551903325ff7,
title = "Accuracy of preimplantation genetic diagnosis in equine in vivo-recovered and in vitro-produced blastocysts",
abstract = "Preimplantation genetic diagnosis has great potential in the horse, but information on evaluation of equine embryo biopsy samples is limited. Blastocysts were biopsied using a Piezo drill and methods for whole-genome amplification (WGA) investigated. Results for 33 genetic loci were then compared between biopsy samples from in vitro-produced (IVP) and in vivo-recovered (VIV) blastocysts. Under the experimental conditions described, WGA using the Qiagen Repli-g Midi kit was more accurate than that using the Illustra Genomiphi V2 kit (98.2{\%} vs 25.8{\%}, respectively). Using WGA with the Qiagen kit, three biopsy samples were evaluated from each of eight IVP and 19 VIV blastocysts, some produced using semen from stallions carrying the genetic mutations associated with the diseases hereditary equine regional dermal asthenia (HERDA), hyperkalemic periodic paralysis (HYPP) or polysaccharide storage myopathy 1 (PSSM1). Three of 81 biopsy samples (3.7{\%}) returned <50{\%} accuracy. In the remaining 78 samples, overall accuracy was 99.3{\%} (2556/2574 loci interrogated). Accuracy did not differ significantly between samples from IVP and VIV blastocysts. Allele drop-out in heterozygous loci was 1.6{\%} (17/1035). Accuracy for sex determination was 100{\%}; accuracy for heterozygosity for disease-causing mutations was 97.7{\%} (43/44). In conclusion, Piezo-driven embryo biopsy with WGA has >95{\%} overall accuracy in IVP and VIV embryos, and this technique is suitable for use in a clinical setting.",
keywords = "embryo biopsy, genetic screening, HERDA, HYPP, polymerase chain reaction (PCR), PSSM, whole genome amplification",
author = "Choi, {Y. H.} and Cecilia Penedo and P. Daftari and Velez, {I. C.} and K. Hinrichs",
year = "2016",
month = "1",
day = "1",
doi = "10.1071/RD14419",
language = "English (US)",
volume = "28",
pages = "1382--1389",
journal = "Reproduction, Fertility and Development",
issn = "1031-3613",
publisher = "CSIRO",
number = "9",

}

TY - JOUR

T1 - Accuracy of preimplantation genetic diagnosis in equine in vivo-recovered and in vitro-produced blastocysts

AU - Choi, Y. H.

AU - Penedo, Cecilia

AU - Daftari, P.

AU - Velez, I. C.

AU - Hinrichs, K.

PY - 2016/1/1

Y1 - 2016/1/1

N2 - Preimplantation genetic diagnosis has great potential in the horse, but information on evaluation of equine embryo biopsy samples is limited. Blastocysts were biopsied using a Piezo drill and methods for whole-genome amplification (WGA) investigated. Results for 33 genetic loci were then compared between biopsy samples from in vitro-produced (IVP) and in vivo-recovered (VIV) blastocysts. Under the experimental conditions described, WGA using the Qiagen Repli-g Midi kit was more accurate than that using the Illustra Genomiphi V2 kit (98.2% vs 25.8%, respectively). Using WGA with the Qiagen kit, three biopsy samples were evaluated from each of eight IVP and 19 VIV blastocysts, some produced using semen from stallions carrying the genetic mutations associated with the diseases hereditary equine regional dermal asthenia (HERDA), hyperkalemic periodic paralysis (HYPP) or polysaccharide storage myopathy 1 (PSSM1). Three of 81 biopsy samples (3.7%) returned <50% accuracy. In the remaining 78 samples, overall accuracy was 99.3% (2556/2574 loci interrogated). Accuracy did not differ significantly between samples from IVP and VIV blastocysts. Allele drop-out in heterozygous loci was 1.6% (17/1035). Accuracy for sex determination was 100%; accuracy for heterozygosity for disease-causing mutations was 97.7% (43/44). In conclusion, Piezo-driven embryo biopsy with WGA has >95% overall accuracy in IVP and VIV embryos, and this technique is suitable for use in a clinical setting.

AB - Preimplantation genetic diagnosis has great potential in the horse, but information on evaluation of equine embryo biopsy samples is limited. Blastocysts were biopsied using a Piezo drill and methods for whole-genome amplification (WGA) investigated. Results for 33 genetic loci were then compared between biopsy samples from in vitro-produced (IVP) and in vivo-recovered (VIV) blastocysts. Under the experimental conditions described, WGA using the Qiagen Repli-g Midi kit was more accurate than that using the Illustra Genomiphi V2 kit (98.2% vs 25.8%, respectively). Using WGA with the Qiagen kit, three biopsy samples were evaluated from each of eight IVP and 19 VIV blastocysts, some produced using semen from stallions carrying the genetic mutations associated with the diseases hereditary equine regional dermal asthenia (HERDA), hyperkalemic periodic paralysis (HYPP) or polysaccharide storage myopathy 1 (PSSM1). Three of 81 biopsy samples (3.7%) returned <50% accuracy. In the remaining 78 samples, overall accuracy was 99.3% (2556/2574 loci interrogated). Accuracy did not differ significantly between samples from IVP and VIV blastocysts. Allele drop-out in heterozygous loci was 1.6% (17/1035). Accuracy for sex determination was 100%; accuracy for heterozygosity for disease-causing mutations was 97.7% (43/44). In conclusion, Piezo-driven embryo biopsy with WGA has >95% overall accuracy in IVP and VIV embryos, and this technique is suitable for use in a clinical setting.

KW - embryo biopsy

KW - genetic screening

KW - HERDA

KW - HYPP

KW - polymerase chain reaction (PCR)

KW - PSSM

KW - whole genome amplification

UR - http://www.scopus.com/inward/record.url?scp=84980349592&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84980349592&partnerID=8YFLogxK

U2 - 10.1071/RD14419

DO - 10.1071/RD14419

M3 - Article

AN - SCOPUS:84980349592

VL - 28

SP - 1382

EP - 1389

JO - Reproduction, Fertility and Development

JF - Reproduction, Fertility and Development

SN - 1031-3613

IS - 9

ER -