Clinical Applications of Cell-Free Fetal DNA From Maternal Plasma

Rijnders, Robbert J.P.; Christiaens, G. C. M. L.; Bossers, Bernadette; Smagt, Jasper J. van der; Schoot, C. Ellen van der; Haas, Masja de

doi:10.1097/01.aog.0000103996.44503.f1

Cited by 89 publications

(53 citation statements)

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“…(97.2%) [24] and Davalieva et al, (89.2%) [25]. This higher sensitivity results could be attributed to longer time range of pregnancy that led to high probability of detecting cffDNA due to the gradual increase of its concentration with the more gestational age [26].…”

Section: Discussionmentioning

confidence: 92%

Comparative Value of Real Time and Conventional PCR in Antenatal Gender Determination

Azab

Salim

Farag

2016

BESPS

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Invasive procedures including chorionic villus sampling and amniocentesis in sexlinked diseases increase the risk of fetal loss. Therefore, Noninvasive fetal gender determination using cell-free fetal DNA (cffDNA) in maternal plasma may be promising. Fifty pregnant females with gestational age ranging from six to ten weeks were included. cffDNA were extracted from maternal plasma and amplified by real time and conventional PCR for identification of SRY, DYS14 and DAZ genes as specific genetic markers for male-bearing pregnancies. In general, sensitivity and specificity of real time PCR was better than conventional PCR. However, sensitivity of DYS14 gene and specificity of SRY gene by real time PCR was equal to those of conventional PCR.Sensitivity of DYS14 gene was the highest and sensitivity of SRY gene was the lowest.However, combination of the three Y-chromosomal sequences in the diagnosis increased the accuracy of the test, which is suitable for clinical application.

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Section: Discussionmentioning

confidence: 92%

Comparative Value of Real Time and Conventional PCR in Antenatal Gender Determination

Azab

Salim

Farag

2016

BESPS

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“…Outra utilidade do exame é o caso de risco de recorrência de hiperplasia congênita de supra-renal quando se usa corticosteroidoterapia como esquema terapêutico profilático logo no inicio da gestação para prevenir a possível virilização dos fetos femininos afetados. Em fetos do sexo masculino, não há necessidade de tratamento, evitando-se assim, o uso desnecessário da medicação que pode gerar efeitos colaterais para a gestante 16,21 . Em conclusão, o diagnóstico do sexo fetal a partir da 5ª semana de gestação pela técnica de PCR em tempo real apresenta boa sensibilidade e excelente especificidade.…”

Section: Discussionunclassified

“…Atualmente, usando-se a técnica de PCR em tempo real, a grande maioria dos trabalhos tem mostrado sensibilidade e especificidade de 100% para o diagnóstico do sexo fetal no primeiro trimestre de gestação [13][14][15][16] . Esses trabalhos, porém, priorizaram a detecção do DNA fetal a partir da 7ª semana de gestação, havendo dúvida se seria possível a detecção em idades gestacionais mais precoces.…”

Section: Introductionunclassified

Determinação precoce do sexo fetal pela análise do DNA no plasma materno

Martinhago¹,

Oliveira²,

Canas³

et al. 2006

Rev. Bras. Ginecol. Obstet.

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RESUMOObjetivo: avaliar a possibilidade do diagnóstico precoce do sexo fetal no plasma materno pela técnica da reação em cadeia da polimerase em tempo real (PCR em tempo real) a partir da 5ª semana de gestação. Métodos: nesse estudo prospectivo foi coletado sangue periférico de gestantes com feto único a partir da 5ª semana de gestação. Após centrifugação do sangue, 0,4 mL de plasma foi separado para extração de DNA fetal. O DNA foi analisado em duplicata por PCR em tempo real para duas regiões genômicas (uma do cromossomo Y e outra comum a ambos os sexos) pelo método de TaqMan ® , o qual utiliza um par de primers e uma sonda fluorescente. Foram excluídos da amostragem os casos que evoluíram para aborto. Para o cálculo da sensibilidade e especificidade, usamos o método de comparação com padrão-ouro, que foi o sexo ao nascimento. Resultados: foram realizados 79 exames de DNA fetal no plasma materno de 52 gestantes. O resultado dos exames foi comparado com o sexo da criança após o parto. O índice de acerto conforme a idade gestacional foi de 92,6% (25 de 27 casos) na 5ª semana, conferindo sensibilidade de 87% e 95,6% (22 de 23 casos) na 6ª semana, com sensibilidade de 92%. A partir da 7ª semana de gestação o acerto foi em 100% (29 de 29 casos). A especificidade foi de 100% independente da idade gestacional. Conclusões: a técnica de PCR em tempo real para detecção do sexo fetal a partir da 5ª semana no plasma materno possui boa sensibilidade e excelente especificidade. Houve concordância do resultado em 100% dos casos em que o diagnóstico foi masculino, independente da idade gestacional, e no caso de feminino, a partir da 7ª semana de gestação. PALAVRAS-CHAVE:Determinação do sexo (Análise); Reação em cadeia da polimerase/métodos; Diagnóstico pré-natal; Idade gestacional ABSTRACT Purpose: to verify the viability of early diagnosis of fetal gender in maternal plasma by the real-time polymerase chain reaction (real-time PCR) starting at the 5th week of pregnancy. Methods: peripheral blood was collected from pregnant women with single fetus starting at the 5th week of gestation. After centrifugation, 0.4 mL plasma was separated for fetal DNA extraction. The DNA was analyzed in duplicate by real-time PCR for two genomic regions, one of the Y chromosome and the other common to both sexes, through the TaqMan ® method, which uses a pair of primers and a fluorescent probe. Patients who aborted were excluded. Results: a total of 79 determinations of fetal DNA in maternal plasma were performed in 52 pregnant women. The results of the determinations were compared to fetal gender after delivery. Accuracy according to gestational age was 92.6% (25 of 27 cases) at 5 weeks with 87% sensitivity, and 95.6% (22 of 23 cases) at 6 weeks with 92% sensitivity. Starting at the 7th week of pregnancy, accuracy was 100% (29 of 29 cases). Specificity was 100% regardless of gestational age. Conclusion: real-time PCR for the detection of fetal gender in maternal plasma starting at the 5th week of gestation has good sensitivity and excellent spe...

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“…Fetal SRY and RHD DNA detection from maternal plasma has reached close to 100% accuracy, as confirmed by many largescale evaluations (6)(7)(8)(9). The high level of diagnostic accuracy is attained by the analytical sensitivity contributed by the use of real-time quantitative PCR (5,10) and the analytical specificity conferred by the choice of fetal DNA targets that are absolutely fetal-specific.…”

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confidence: 88%

MS analysis of single-nucleotide differences in circulating nucleic acids: Application to noninvasive prenatal diagnosis

Ding

Chiu

Lau

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

177

152

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The analysis of circulating nucleic acids has revealed applications in the noninvasive diagnosis, monitoring, and prognostication of many clinical conditions. Circulating fetal-specific sequences have been detected and constitute a fraction of the total DNA in maternal plasma. The diagnostic reliability of circulating DNA analysis depends on the fractional concentration of the targeted sequence, the analytical sensitivity, and the specificity. The robust discrimination of single-nucleotide differences between circulating DNA species is technically challenging and demands the adoption of highly sensitive and specific analytical systems. We have developed a method based on single-allele base extension reaction and MS, which allows for the reliable detection of fetal-specific alleles, including point mutations and single-nucleotide polymorphisms, in maternal plasma. The approach was applied to exclude the fetal inheritance of the four most common Southeast Asian ␤-thalassemia mutations in at-risk pregnancies between weeks 7 and 21 of gestation. Fetal genotypes were correctly predicted in all cases studied. Fetal haplotype analysis based on a single-nucleotide polymorphism linked to the ␤-globin locus, HBB, in maternal plasma also was achieved. Consequently, noninvasive prenatal diagnosis in a mother and father carrying identical ␤-thalassemia mutations was accomplished. These advances will help in catalyzing the clinical applications of fetal nucleic acids in maternal plasma. This analytical approach also will have implications for many other applications of circulating nucleic acids in areas such as oncology and transplantation. R ecently, much interest has been focused on the biology and diagnostic applications of nucleic acids that are present in the plasma and serum of humans (1, 2). In particular, fetal DNA has been found to exist in maternal plasma (3). This discovery has facilitated the development of noninvasive prenatal diagnostic approaches based simply on the analysis of a maternal blood sample (4). The noninvasive nature of maternal plasmabased approaches represents a major advantage over conventional methods of prenatal diagnosis, such as amniocentesis and chorionic villus sampling, which are associated with a small but finite risk of fetal loss. However, a technical challenge experienced by many workers in the field relates to the ability to discriminate fetal DNA from the coexisting background of maternal DNA in maternal plasma. During pregnancy, fetal DNA amounts to Ϸ3-6% of the total DNA in maternal plasma (5). Hence, the diagnostic reliability of fetal DNA analysis in maternal plasma depends on the sensitivity and specificity of the analytical system for the detection of fetal-specific markers.Fetal SRY and RHD DNA detection from maternal plasma has reached close to 100% accuracy, as confirmed by many largescale evaluations (6-9). The high level of diagnostic accuracy is attained by the analytical sensitivity contributed by the use of real-time quantitative PCR (5, 10) and the analytical specifici...

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Clinical Applications of Cell-Free Fetal DNA From Maternal Plasma

Abstract: Detection of cell-free fetal DNA from maternal plasma is a reliable technique that can substantially reduce invasive prenatal tests.

Cited by 89 publications

References 20 publications

Comparative Value of Real Time and Conventional PCR in Antenatal Gender Determination

Comparative Value of Real Time and Conventional PCR in Antenatal Gender Determination

Determinação precoce do sexo fetal pela análise do DNA no plasma materno

MS analysis of single-nucleotide differences in circulating nucleic acids: Application to noninvasive prenatal diagnosis

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