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Prenatal diagnosis of genetic diseases directly using paper-dried cord blood as the starting material for PCR

  • Huan HuangEmail author
  • You Zhou
  • Jiajia Zhang
  • Weiyin Yao
  • Guoying ZhangEmail author
Research Paper

Abstract

A rapid and low-cost method of diagnosis is becoming important for detecting fetal inherited diseases, including single-gene disorders and chromosomal abnormalities. Here, we demonstrated an innovation that use paper-dried cord blood (PCB) as the starting material for PCR and whole genome amplification without any DNA extraction step at a very low cost. A novel PCR buffer named “DDB buffer” containing ammonium sulfate and glycerol were used instead of the conventional 10× PCR buffer. The amplicons were directly analyzed through microchip electrophoresis and whole genome sequencing. Inhibitory substances in filter paper were effectively inactivated using DDB buffer. Direct PCR amplification of DNA fragments ranging from 100 to 900 bp using filter paper spotted with 0.5 to 5 μL of cord blood and various anticoagulants was successful. We were able to determine fetal single-gene disorders and chromosomal diseases in all 46 chromosomes using PCB samples successfully. Compared with prenatal diagnosis using purified DNA, the proposed method is simple, fast, less prone to cross-contamination at minimal cost. Researchers and clinical and healthcare workers may employ this method for genetic diagnosis using cord blood samples with minimum laboratory resources. This method is very promising for a variety of genetic diagnosis applications in underserved communities at the point of need in developing areas.

Graphical abstract

Keywords

Direct PCR Paper-dried cord blood Prenatal diagnosis Single-gene disorders Chromosomal abnormalities 

Abbreviations

COL1A2

Collagen type 1 alpha 2

DD

Developmental delay

DMD

Duchenne muscular dystrophy

ID

Intellectual disability

IDS

Iduronate-2-sulfatase

PAH

Phenylalanine hydroxylase

PCB

Paper-dried cord blood

PCR

Polymerase chain reaction

SMN1

Survival of motor neuron 1

Notes

Funding information

This work was supported by the National Natural Science Foundation of China (81741040), Medical Talent of Empowering Medicine through Science and Education Program (QNRC2016618), the Key Provincial Talents of Maternal and Child Health Program in Jiangsu Province (FRC2017-43), and the project for the Priority Academic Program Development of Jiangsu Higher Education Institutions (JX10231802). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

216_2019_2048_MOESM1_ESM.pdf (153 kb)
ESM 1 (PDF 152 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Obstetrics and GynecologyThe First Affiliated Hospital of Nanjing Medical UniversityNanjingChina
  2. 2.Department of Department of Cell Systems & AnatomyUniversity of Texas Health Science Center at San AntonioSan AntonioUSA

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