Assessment of a Simplified Cell-Free DNA Method for Prenatal Down Syndrome Screening

• Published in: Clinical chemistry (Baltimore, Md.) Md.), 2022-11, Vol.68 (11), p.1449-1458
• Main Authors: Palomaki, Glenn E, Eklund, Elizabeth E, Kloza, Edward M, Lambert-Messerlian, Geralyn M
• Format: Article
• Language: English
• Subjects: Cell-Free Nucleic Acids; Down Syndrome - diagnosis; Down Syndrome - genetics; Female; Humans; Pregnancy; Prenatal Diagnosis - methods; Trisomy; Trisomy 13 Syndrome - diagnosis
• ISSN: 0009-9147; 1530-8561
• DOI: 10.1093/clinchem/hvac131

Prenatal screening for common trisomies via cell-free (cfDNA) is usually implemented by technologies utilizing massively parallel sequencing, stringent environmental controls, complex bioinformatics, and molecular expertise. An alternative and less complex methodology utilizes rolling circle amplification (RCA). Further evaluation of its performance and related requirements are warranted. At 16 sites, women at 10 to 20 weeks gestation provided informed consent, relevant information, and 2 to 3 blood samples. Samples shipped for testing were processed and stored. Women were enrolled at primary cfDNA screening, or following such screening at referral for diagnostic testing. RCA testing occurred post-enrollment, over 11 months. Diagnostic results and delivery notes determined clinical truth. Detection rates were based on confirmed trisomic pregnancies; false-positive rates were based on unaffected pregnancies from the general population. Detection rate for the common trisomies was 95.9% (117/122, 95% CI, 90.5%-98.5%); overall false-positive rate was 1.00% (22/2,205, 0.65%-1.51%). Test failure rate after repeat testing was 0.04%. When assay standard deviations were below pre-specified levels, the overall false-positive rate was much lower at 0.30% (P < 0.001). Fetal sex calls were correct for 99.7%. One technician analyzed 560 samples over 2 weeks, a rate of 14 000/year. Our assessment of this simplified cfDNA-based system for prenatal screening for common trisomies performed in a prenatal screening laboratory is encouraging. Improved detection, low failure rates and rapid reporting can be achieved by collecting 2 samples. Future priorities should include achieving higher run precision using a single collection tube. NCT03087357.