title: [Application study of droplet digital PCR to detect maternal cell contamination in prenatal diagnosis].
abstract: Objective:To develop a new method based on droplet digital PCR (DD-PCR) for detection and quantification of maternal cell contamination in prenatal diagnosis.Methods:Invasive prenatal samples from 40 couples of β(IVS-Ⅱ-654)/β(N) thalassemia gene carriers who accepted prenatal diagnosis in Affiliated Women and Children's Hospital of Guangzhou Medical University from October 2015 to December 2016 were analyzed retrospectively. Specific primers and probes were designed. The concentration gradient were 50%, 25%, 12.5%, 6.25%, 3.125%, 1.562 5%. There were 40 groups of prenatal diagnostic samples. Comparing DD-PCR with quantitative fluorescent-PCR (QF-PCR) based on the short tandem repeats for assement of the sensitivity and accuracy of maternal cell contamination, respectively.Results:DD-PCR could quantify the maternal cell contamination as low as 1.562 5%. The result was proportional to the dilution titers. In the 40 prenatal samples, 6 cases (15%, 6/40) of maternal cell contamination were detected by DD-PCR, while the QF-PCR based on short tandem repeat showed 3 cases (7.5%, 3/40) with maternal cell contamination, DD-PCR was more accurate (P=0.002) .Conclusion:DD-PCR is a precise and sensitive method in the detection of maternal cell contamintation. It could be useful in clinical application.
authors.affiliation: Prenatal Diagnosis Center, Guangdong Women and Children's Hospital, Guangzhou Medical University. Guangzhou 511400, China.
authors.forename: J
authors.initials: J
authors.lastname: Geng
authors.forename: C
authors.initials: C
authors.lastname: Liu
authors.forename: X C
authors.initials: XC
authors.lastname: Zhou
authors.forename: J
authors.initials: J
authors.lastname: Ma
authors.forename: L
authors.initials: L
authors.lastname: Du
authors.forename: J
authors.initials: J
authors.lastname: Lu
authors.forename: W N
authors.initials: WN
authors.lastname: Zhou
authors.forename: T T
authors.initials: TT
authors.lastname: Hu
authors.forename: L J
authors.initials: LJ
authors.lastname: Lyu
authors.forename: A H
authors.initials: AH
authors.lastname: Yin
mesh_terms: D004247:DNA / Q000032:analysis*; D017931:DNA Primers; D005260:Female; D006801:Humans; D062145:Maternal Serum Screening Tests / Q000379:methods*; D018895:Microsatellite Repeats / Q000235:genetics; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013789:Thalassemia
chemical_list: D017931:DNA Primers; D004247:DNA
keywords: DNA contamination; Genotype; Polymerase chain reaction; Prenatal diagnosis; Thalassemia