Novel method of real-time PCR-based screening for common fetal trisomies

BMC Medical Genomics

Ji Hyae Lim¹  Hyun Mee Ryu¹  Sohee Oh²  Chan-Wook Park³  Sun Min Kim³  Seung Mi Lee³  Byoung Jae Kim³  Joong Shin Park³  Jong Kwan Jun³  So Yeon Kim³  Ig Hwan Oh⁴  Ja Nam Koo⁴ 

[1] Center for Prenatal Biomarker Research, CHA Advanced Research Institute;Department of Biostatistics, Seoul Metropolitan Government Seoul National University Boramae Medical Center;Department of Obstetrics and Gynecology, Seoul National University College of Medicine;Seoul Women’s Hospital;

关键词: Fetal trisomy;    Prenatal diagnosis;    Non-invasive prenatal test;    Real-time polymerase chain reaction;    Peptide nucleic acid;   

DOI  :  10.1186/s12920-021-01039-1

来源: DOAJ

【 摘 要 】

Abstract Background The non-invasive prenatal test (NIPT) is based on next generation sequencing (NGS) and is used for screening for fetal trisomy. However, it is time-consuming and technically difficult. Recently, peptide nucleic acid (PNA) probe-based real-time polymerase chain reaction (RT-PCR) was developed. This study aimed to examine the performance of the RT-PCR-based NIPT for screening of common fetal trisomies Methods From stored maternal plasma, RT-PCR was performed using Patio™ NIPT Detection Kit. In melting curve analysis, the height of melting peaks of target chromosome and reference chromosome was calculated as a peak ratio. The adjusted peak ratio of 8 markers with correction factors in each target chromosome was summated and calculated to z-score. The cut-off value for each target chromosome was established for classification (low risk vs. high risk for trisomy) whose performance was obtained in the validation phase. Results 330 plasma samples from pregnant women with normal fetus and 22 trisomy cell-line samples were used to establish the optimal cut-off values for z-score of each target chromosome. In the validation phase, 1023 samples from pregnant women including 22 cases with fetal trisomy and 1001 cases of normal control were used. The RT-PCR-based NIPT showed 95.45% sensitivity [95% confidence interval (CI) 77.16–99.88%], 98.60% specificity (95% CI 97.66–99.23%), and 98.53% accuracy (95% CI 97.59–99.18%) for the identification of trisomy 21, 18, or 13. Of 1023 samples, fifteen cases were mismatched for classification [one case as a false negative (false negative rate: 4.5%) and 14 cases as false positives (false positive rate: 1.4%)]. Conclusion The RT-PCR-based NIPT showed high sensitivity and specificity for the detection of common fetal trisomies and it could be a feasible alternative to NGS-based NIPT.

【 授权许可】

Unknown