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Related Experiment Videos

Prenatal RHD genotyping in Croatia: preliminary results.

J Bingulac-Popović1, I Babić1, V Đogić1

  • 1Department of Molecular Diagnostics, Croatian Institute of Transfusion Medicine, Petrova 3, 10000 Zagreb, Croatia.

Transfusion Clinique Et Biologique : Journal De La Societe Francaise De Transfusion Sanguine
|November 23, 2020
PubMed
Summary

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This summary is machine-generated.

Non-invasive fetal RHD genotyping using RT-PCR accurately determines fetal RhD status, enabling targeted anti-D prophylaxis for RhD-negative mothers. This reliable method ensures high diagnostic accuracy, improving antenatal care.

Area of Science:

  • Medical Diagnostics
  • Genetics
  • Immunology

Background:

  • The Croatian Institute of Transfusion Medicine (CITM) sought to implement non-invasive fetal RHD genotyping for targeted antenatal anti-D prophylaxis.
  • Accurate determination of fetal RHD status is crucial for managing RhD incompatibility in pregnancy.

Purpose of the Study:

  • To analyze the diagnostic performance of an in-house RT-PCR method for fetal RHD genotyping.
  • To evaluate preliminary results of fetal RHD genotyping in RhD-negative pregnant women.

Main Methods:

  • Retrospective analysis of RHD genotyping results from 205 RhD-negative pregnant women (12-36 weeks gestation) between 2015-2020.
  • Automated cell-free DNA (cffDNA) extraction using QIAsymphony SP DSP Virus Midi Kit.
  • RT-PCR amplification of RHD exons 5, 7, and 10, with SRY, RASSF1A, and β-actin genes as internal controls.
Keywords:
ADN fœtal acellulaireAnti-D immunoprophylaxisCell-free fetal DNAGénotypage prénatal fœtal RHD non invasifHemolytic disease of fetus and newbornImmunoprophylaxie de l’immunisation fœto-maternelle RhésusDMaladie hémolytique du nouveau-néNon-invasive prenatal RHD genotyping

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Main Results:

  • Identified fetal RHD genotypes in 70.72% positive and 28.78% negative cases.
  • Achieved 100% sensitivity, 95.08% specificity, and 98.48% diagnostic accuracy.
  • Observed no false negatives and only 1.50% false positives, with 100% negative and 97.86% positive predictive values.

Conclusions:

  • Automated cffDNA extraction and RT-PCR offer a highly reliable system for fetal RHD status determination.
  • This method enables targeted antenatal anti-D prophylaxis, improving pregnancy management.
  • Strict cleaning procedures are essential for high specificity in cffDNA extraction.