Seung Yong Lee1, Seung Jun Kim1, Sung-Hee Han1, Joon Soo Park1, Hyo Jung Choi1, Jeong Jin Ahn1, Moon-Ju Oh1, Sung Han Shim2, Dong Hyun Cha3, Seung Yong Hwang4. 1. BioCore Co. Ltd., Division of Biotechnology, Guro-gu, Seoul, Republic of Korea. 2. Department of Obstetrics and Gynecology, CHA Gangnam Medical Center, CHA University, Seoul, Republic of Korea. 3. Department of Obstetrics and Gynecology, CHA Gangnam Medical Center, CHA University, Seoul, Republic of Korea; Genetics Laboratory, Fertility Center of CHA Gangnam Medical Center, CHA University, Seoul, Republic of Korea. Electronic address: chadh001@chamc.co.kr. 4. BioCore Co. Ltd., Division of Biotechnology, Guro-gu, Seoul, Republic of Korea; Department of Bio-Nanotechnology, Hanyang University, Sangnok-gu, Ansan, Gyeonggi-do, Republic of Korea; Department of Molecular and Life Science, Hanyang University, Sangnok-gu, Ansan, Gyeonggi-do, Republic of Korea. Electronic address: syhwang@bio-core.com.
Abstract
BACKGROUND: Non-invasive prenatal screening (NIPS) of trisomy 21 (T21) using digital PCR (dPCR) with several advantages will be very effective. Here, we developed a dPCR system for T21 screening which allows high sensitivity and real-time diagnosis and thus overcome sequence based analysis. METHODS: Cut-off value was established using DNA extracted from all 157 T21 negative samples including 47 pregnant woman samples and 3 T21 positive pregnant woman samples extracted from 4 different sample types. To increase the portion of the cell-free fetal DNA (cffDNA) in maternal cell-free DNA (cfDNA), a size selection method was devised. We evaluated the clinical reliability of NIPS using dPCR through analysis of 877 pregnant woman samples. RESULTS: We could demonstrate the possibility of NIPS using dPCR performed by applying cut-off value and size selection method. The overall accuracy was derived at 99.66% using 877 pregnant woman plasma samples. CONCLUSION: Our results showed that dPCR can meet the requirements for NIPS for T21. It is relatively inexpensive, easy to use in a screening method and compatible with ethical concerns regarding access to nucleotide sequence information. This study may be a basic data for the development of population-wide screening for T21 in pregnant women.
BACKGROUND: Non-invasive prenatal screening (NIPS) of trisomy 21 (T21) using digital PCR (dPCR) with several advantages will be very effective. Here, we developed a dPCR system for T21 screening which allows high sensitivity and real-time diagnosis and thus overcome sequence based analysis. METHODS: Cut-off value was established using DNA extracted from all 157 T21 negative samples including 47 pregnant woman samples and 3 T21 positive pregnant woman samples extracted from 4 different sample types. To increase the portion of the cell-free fetal DNA (cffDNA) in maternal cell-free DNA (cfDNA), a size selection method was devised. We evaluated the clinical reliability of NIPS using dPCR through analysis of 877 pregnant woman samples. RESULTS: We could demonstrate the possibility of NIPS using dPCR performed by applying cut-off value and size selection method. The overall accuracy was derived at 99.66% using 877 pregnant woman plasma samples. CONCLUSION: Our results showed that dPCR can meet the requirements for NIPS for T21. It is relatively inexpensive, easy to use in a screening method and compatible with ethical concerns regarding access to nucleotide sequence information. This study may be a basic data for the development of population-wide screening for T21 in pregnant women.