title: Primer-introduced restriction analysis polymerase chain reaction method for non-invasive prenatal testing of β-thalassemia.
abstract: We have developed a new method for non-invasive prenatal testing (NIPT) of paternally inherited fetal mutants for β-thalassemia (β-thal). Specially designed primer-introduced restriction analysis-polymerase chain reaction (PIRA-PCR) were used to detect four major mutations [IVS-II-654, HBB: c.316-197C > T; codon 17 (A > T), HBB: c.52A > T; -28 (A > G), HBB: c.-78A > G and codons 41/42 (-TTCT), HBB: c.126_129delCTTT] causing β-thal in China. The PIRA-PCR assay was first tested in a series of mixed DNA with different concentrations and mixed proportions. Subsequently, this assay was further tested in 10 plasma DNA samples collected from pregnant women. In the DNA mixture simulation test, the PIRA-PCR assay was able to detect 3.0% target genomic DNA (gDNA) mixed in 97.0% wild-type gDNA isolated from whole blood. For plasma DNA testing, the results detected by PIRA-PCR assay achieved 100.0% consistency with those obtained from the amniocentesis analysis. This new method could potentially be used for NIPT of paternally inherited fetal mutants for β-thal.
authors.affiliation: Department of Science and Technology, BGI-Shenzhen , Shenzhen , People's Republic of China .
authors.forename: Saijun
authors.initials: S
authors.lastname: Liu
authors.forename: Liyuan
authors.initials: L
authors.lastname: Chen
authors.forename: Xiandong
authors.initials: X
authors.lastname: Zhang
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.forename: Haiying
authors.initials: H
authors.lastname: Lin
authors.forename: Louhui
authors.initials: L
authors.lastname: Liu
authors.forename: Jiansheng
authors.initials: J
authors.lastname: Xie
authors.forename: Huijuan
authors.initials: H
authors.lastname: Ge
authors.forename: Minglan
authors.initials: M
authors.lastname: Ye
authors.forename: Caifen
authors.initials: C
authors.lastname: Chen
authors.forename: Xingwen
authors.initials: X
authors.lastname: Ji
authors.forename: Caifen
authors.initials: C
authors.lastname: Zhang
authors.forename: Fengping
authors.initials: F
authors.lastname: Xu
authors.forename: Hui
authors.initials: H
authors.lastname: Jiang
authors.forename: Hefu
authors.initials: H
authors.lastname: Zhen
authors.forename: Shiping
authors.initials: S
authors.lastname: Chen
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
mesh_terms: D001483:Base Sequence; D004252:DNA Mutational Analysis / Q000379:methods; D017931:DNA Primers / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D055544:beta-Globins / Q000235:genetics*; D017086:beta-Thalassemia / Q000175:diagnosis / Q000235:genetics*
chemical_list: D017931:DNA Primers; D055544:beta-Globins
keywords: non-invasive prenatal testing (NIPT); primer-introduced restriction analysis-polymerase chain reaction (PIRA-PCR); β-Thalassemia (β-thal)