Development of noninvasive prenatal testing of down’s syndrome by droplet digital PCR

Authors

  • Narutchala Suwannakhon School of Science, University of Phayao
  • Tanapat Pangeson School of Medical Science, University of Phayao
  • Arunee Pingyod Thalassaemia Unit, University of Phayao
  • Khwanruedee Mahingsa Thalassaemia Unit, University of Phayao
  • Wanwipa Bumrungpakdee Thalassaemia Unit, University of Phayao
  • Torpong Sanguansermsri Thalassaemia Unit, University of Phayao

Keywords:

Down’s syndrome, Prenatal diagnosis, NIPT, ddPCR

Abstract

Down’s syndrome caused by aneuploidy of chromosome 21 (trisomy 21: T21) has high prevalence. Nowadays, advanced DNA technologies are used in noninvasive prenatal testing (NIPT) to screen for numerical chromosomal abnormalities in pregnancy at advanced maternal age. These technologies are expensive and complicated. The aim of this study is to develop the droplet digital PCR (ddPCR) technique for NIPT of T21. DdPCR condition was optimized by using DNA samples extracted from blood of children with T21, blood and plasma of non-pregnant normal women. Then, plasma cell-free DNA (cfDNA) from 30 pregnant women was tested using optimized ddPCR. Ratio of mutant allele/wild type allele was calculated by Bio-Rad QuantaSoft Analysis program. The ratios of blood and plasma of non-pregnant women were 1.043±0.073 and 0.710±0.019, respectively. The ratio of T21 patients was 1.486±0.107. The ratio of negative control plasma samples was 0.705±0.074. The result of cfDNA from plasma of 29 pregnant women was 0.742±0.062 which meant that there was a low probability to have T21 fetuses. The results of NIPT in 29 cfDNA samples were the same as those of cytogenetic methods. One cfDNA sample was excluded because of vanishing twin syndrome. Thus, this developed ddPCR-based NIPT was able to detect a low-risk group for T21; however, more pregnancy plasma samples are needed in order to find pregnant women with high risk of having T21 fetuses.

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ตัวอย่างผลจาก ddPCR ของตัวอย่างดีเอ็นเอจากพลาสมาหญิงตั้งครรภ์ที่มีฟีตัสปกติโดยแสดงอัตราส่วนและจำนวนที่ให้ผลบวกต่อโพรบสำหรับ mutant allele และ wild type allele จากโปรแกรม Bio-Rad QuantaSoft Analysis

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Published

2023-08-27

How to Cite

1.
Suwannakhon N, Pangeson T, Pingyod A, Mahingsa K, Bumrungpakdee W, Sanguansermsri T. Development of noninvasive prenatal testing of down’s syndrome by droplet digital PCR. Health Sci Tech Rev [Internet]. 2023 Aug. 27 [cited 2024 May 16];16(2):40-52. Available from: https://li01.tci-thaijo.org/index.php/journalup/article/view/258697

Issue

Vol. 16 No. 2 (2023): May - August

Section

Research articles

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