Non‐Invasive prenatal testing with rolling circle amplification: Real‐world clinical experience in a non‐molecular laboratory

Abstract

Background Non‐invasive prenatal testing (NIPT) using cell‐free DNA (cfDNA) circulating in maternal blood provides a sensitive and specific screening technique for common fetal aneuploidies, but the high cost and workflow complexity of conventional methodologies limit its widespread implementation. A unique rolling circle amplification methodology reduces cost and complexity, providing a promising alternative for increased global accessibility as a first‐tier test. Methods In this clinical study, 8160 pregnant women were screened on the Vanadis system for trisomies 13, 18, and 21, and positive results were compared to clinical outcomes where available. Results The Vanadis system yielded a 0.07% no‐call rate, a 98% overall sensitivity, and a specificity of over 99% based on available outcomes. Conclusion The Vanadis system provided a sensitive, specific, and cost‐effective cfDNA assay for trisomies 13, 18, and 21, with good performance characteristics and low no‐call rate, and it eliminated the need for either next‐generation sequencing or polymerase chain reaction amplification.

Technical Advances in Circulating Cell-Free DNA Detection and Analysis for More Personalized Medicine in Patients’ Care

“…In VANADIS assay (PerkinElmer, Waltham, MA), DNA fragments are labeled with fluorescent oligonucleotides specific for precise genetic targets, then circularized and copied multiple times before being placed on a 96-well nanofilter microplate and analyzed by imaging [106]. This assay is now applied to prenatal screening with high accuracy [107,108]. Since this method does not require DNA amplification and sequencing, is easily implemented in any laboratory, scalable and fully automated.…”

Technical Advances in Circulating Cell-Free DNA Detection and Analysis for Personalized Medicine in Patients’ Care

“…In the VANADIS assay (PerkinElmer, Waltham, MA), DNA fragments are labeled with fluorescent oligonucleotides specific for precise genetic targets, then circularized and copied multiple times before being placed on a 96-well nanofilter microplate and analyzed by imaging [82]. This assay is now applied to prenatal screening with high accuracy [83,84]. Since this method does not require DNA amplification and sequencing, it is easily implemented in any laboratory, scalable, and fully automated.…”

Noninvasive Prenatal Testing Test Versus Chorionic Villus Sampling, Where Are We Now?

“…However, a new methodology using rolling circle replication has reduced the no-call rate to 0.07% with an overall sensitivity of 98% and overall specificity of 99% for trisomy 21, 18, and 13. 17 A no-call result may indicate a higher chance of atypical chromosomal abnormalities, adverse pregnancy outcomes, and maternal neoplasia such as lymphoma.…”