QF-PCR as a substitute for karyotyping of cytotrophoblast for the analysis of chorionic villi: advantages and limitations from a cytogenetic retrospective audit of 44,727 first-trimester prenatal diagnoses

Prenat Diagn. 2013 May;33(5):502-8. doi: 10.1002/pd.4099. Epub 2013 Apr 21.

Abstract

Objectives: Karyotyping on chorionic villous samples (CVS) includes the analysis of both cytotrophoblast (STC) and mesenchyme (LTC). This approach requires complex laboratory organization and trained technicians. The introduction of quantitative fluorescent polymerase chain reaction (QF-PCR) instead of conventional karyotyping in low-risk pregnancies opened its application in CVS analysis. Discordant QF-PCR and CVS cytogenetic results were reported, and strategies for CVS analysis were introduced to minimize this risk. The possibility to substitute the STC with QF-PCR was reported. The aim of this study is to evaluate benefits and limitations of the approach QF-PCR + LTC compared with the traditional method STC + LTC and to quantify the associated risks of false results.

Method: This study is based on a retrospective cytogenetic audit of CVS results (n = 44 727) generated by the STC + LTC analytic approach. False-negative risks related to true fetal mosaicism type IV, imprinting syndromes and maternal contamination in LTC were calculated.

Results: Compared with STC + LTC, QF-PCR + LTC approach is associated with a cumulative false-negative risk of ~1/3100-1/4400. Costs and reporting time of STC in a high-throughput cytogenetic lab are similar to a CE-IVD marked QF-PCR analysis.

Conclusions: These results should be clearly highlighted in the pre-test counseling and extensively discussed with the couple prior to testing for informed consent.

Publication types

  • Evaluation Study

MeSH terms

  • Chorionic Villi Sampling / statistics & numerical data
  • Chorionic Villi*
  • Chromosome Aberrations / statistics & numerical data
  • Clinical Audit
  • Cost-Benefit Analysis
  • Female
  • Fluorescence
  • Humans
  • Karyotyping / economics
  • Karyotyping / methods
  • Limit of Detection
  • Polymerase Chain Reaction / economics
  • Polymerase Chain Reaction / methods*
  • Predictive Value of Tests
  • Pregnancy
  • Pregnancy Trimester, First*
  • Prenatal Diagnosis / economics
  • Prenatal Diagnosis / methods*
  • Retrospective Studies
  • Trophoblasts*