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An improved method to extract DNA from 1 ml of uncultured amniotic fluid from patients at less than 16 weeks' gestation.

ABSTRACT: The aim of this study was to develop an improved technique for DNA extraction from 1 ml of uncultured AF from patients with a gestational age less than 16 weeks and to allow the use of array-CGH without DNA amplification. The DNA extraction protocol was tested in a series of 90 samples including 41 of uncultured AF at less than 16 weeks of gestation. Statistical analyses were performed using linear regression. To evaluate the sensitivity and the specificity of array-CGH on 1 ml of uncultured AF, five samples with an abnormal karyotype (three with aneuploidy, two with structural abnormalities) and five with a normal karyotype were studied. This protocol was reproducible and we were able to show a great improvement with higher yield of DNA obtained from all patients, including those with a gestational age less than 16 weeks (p = 0.003). All chromosomal abnormalities were detected and characterized by array-CGH and normal samples showed normal profiles. This new DNA extraction protocol associated with array-CGH analysis could be used in prenatal testing even when gestational age is less than 16 weeks, especially in cases with abnormal ultrasound findings.

SUBMITTER: Mosca-Boidron AL

PROVIDER: S-EPMC3614959 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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An improved method to extract DNA from 1 ml of uncultured amniotic fluid from patients at less than 16 weeks' gestation.

Mosca-Boidron Anne-Laure AL Faivre Laurence L Aho Serge S Marle Nathalie N Truntzer Caroline C Rousseau Thierry T Ragon Clémence C Payet Muriel M Thauvin-Robinet Christelle C Thevenon Julien J El Chehadeh Salima S Huet Fréderic F Sagot Paul P Mugneret Francine F Callier Patrick P

PloS one 20130402 4

The aim of this study was to develop an improved technique for DNA extraction from 1 ml of uncultured AF from patients with a gestational age less than 16 weeks and to allow the use of array-CGH without DNA amplification. The DNA extraction protocol was tested in a series of 90 samples including 41 of uncultured AF at less than 16 weeks of gestation. Statistical analyses were performed using linear regression. To evaluate the sensitivity and the specificity of array-CGH on 1 ml of uncultured AF, ...[more]

PMID: 23565177

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Project description:ObjectiveTo establish a normal reference interval for amniotic sac diameter (ASD) between 7 + 0 and 9 + 6 weeks' gestation and its relative size in relation to gestational sac diameter (GSD) and the embryo crown-rump length (CRL).MethodsThis was a prospective, cross-sectional study of consecutive women presenting to the Early Pregnancy Unit, University College Hospital, London, UK, between August 2022 and June 2023. We included live, normally sited, singleton pregnancies with a normal 20-week anomaly scan. We collected 120 cases per gestational week, from 7 + 0 to 9 + 6 weeks' gestation, totaling 360 cases. We performed an inter- and intraobserver variability assessment in the measurement of mean ASD in 30 patients. Regression analyses were used to establish reference intervals for GSD and CRL, ASD and CRL, GSD and ASD, and GSD/ASD ratio and CRL. A fitted regression line was calculated, along with a 90% prediction interval and R2 value.ResultsThere was good interobserver agreement (mean ± SD difference, 0.007 ± 1.105 mm (95% limits of agreement (LoA), -2.160 to 2.174 mm)) and good intraobserver agreement for Observer A (mean ± SD difference, -0.080 ± 0.741 mm (95% LoA, -1.532 to 1.372 mm)) and Observer B (mean ± SD difference, -0.014 ± 0.919 mm (95% LoA, -1.814 to 1.786 mm)) in the measurement of mean ASD. Regression analyses showed a statistically significant association between each pair of values (P < 0.001 for all). There was a significant quadratic association between mean GSD and CRL (R2 = 56%), mean GSD and ASD (R2 = 60%) and GSD/ASD ratio and CRL (R2 = 68%), and a significant cubic association between mean ASD and CRL (R2 = 90%). The regression equations were used to quantify the values of ASD and GSD/ASD ratios for a range of CRL values and gestational ages.ConclusionOur study has produced comprehensive reference intervals for amniotic sac size in early pregnancy, which could be used in routine clinical practice. © 2024 The Author(s). Ultrasound in Obstetrics & Gynecology published by John Wiley & Sons Ltd on behalf of International Society of Ultrasound in Obstetrics and Gynecology.

Project description:ImportanceThe inclusion of less invasive surfactant administration (LISA) in the care of preterm infants has been found to be beneficial for respiratory outcomes. Recently, the OPTIMIST trial found higher mortality rates in the subgroup of infants born at 25 to 26 weeks' gestational age (GA) who received surfactant treatment while spontaneously breathing.ObjectiveTo analyze outcomes among LISA-exposed, highly vulnerable babies born at less than 27 weeks' GA within the large-scale observational cohort of the German Neonatal Network.Design, setting, and participantsIn this cohort study of data from 68 tertiary level neonatal intensive care units in Germany of infants born between 22 weeks 0 days to 26 weeks 6 days of gestation between April 1, 2009, and December 31, 2020, short-term outcomes among infants receiving LISA vs infants not receiving LISA were compared.ExposureUse of LISA within the first 72 hours of life.Main outcomes and measuresThe main outcomes were rates of LISA use, use of mechanical ventilation within the first 72 hours (considered failure of LISA), and association of LISA with outcomes, including death from all causes, bronchopulmonary dysplasia (BPD), death and BPD combined, pneumothorax, retinopathy of prematurity, intracerebral hemorrhage, and periventricular leukomalacia. To address potential confounding factors, multivariate logistic regression models were used.ResultsA total of 6542 infants (3030 [46.3%] female and 3512 [53.7%] male; mean [SD] GA, 25.3 (1.1) weeks; mean [SD] birth weight, 715 [180] g) were analyzed; 2534 infants (38.7%) received LISA, which was most frequently given quasi-prophylactically during delivery room management. Among the infants who received LISA, 1357 (53.6%) did not require mechanical ventilation in the first 72 hours compared with 331 infants (8.3%) of 4008 who did not receive LISA. In a multivariate logistic regression model that adjusted for GA, small-for-GA status, sex, multiple birth, inborn status, antenatal steroid use, and maximum fraction of inspired oxygen in the first 12 hours of life, LISA was associated with reduced risks of all-cause death (odds ratio [OR], 0.74; 95% CI, 0.61-0.90; P = .002), BPD (OR, 0.69; 95% CI, 0.62-0.78; P < .001), and BPD or death (OR, 0.64; 95% CI, 0.57-0.72; P < .001) compared with infants without LISA exposure.Conclusions and relevanceThe results of this long-term multicenter cohort study suggest that LISA may be associated with reduced risks of adverse outcomes in extremely preterm infants.

Project description:ImportancePrevious research has examined outcomes among very preterm newborns by the birthing parent's race and ethnicity, but knowledge about these trends during the COVID-19 pandemic is limited.ObjectiveTo examine trends in outcomes among Black, Hispanic, and Asian preterm newborns compared with White preterm newborns.Design, setting, and participantsThis cohort study (2018-2022) took place at 774 neonatal intensive care units in the Vermont Oxford Network. Participants were newborns born at 22 to 29 weeks' gestation.ExposureRace and ethnicity.Main outcomes and measuresThe primary outcomes were mortality and complications, including respiratory distress syndrome, necrotizing enterocolitis (NEC), early-onset sepsis, late-onset sepsis (LOS), severe intraventricular hemorrhage (sIVH), severe retinopathy of prematurity, chronic lung disease, pneumothorax, and complication-free survival.ResultsAmong 90 336 newborns (47 215 male [52.3%]; 43 121 female [47.7%]; mean [SD] gestational age, 26.4 [2.1] weeks), 4734 (5.2%) were born to Asian, 20 345 (22.3%) to Hispanic, 31 264 (34.3%) to non-Hispanic Black, and 33 993 (37.3%) to non-Hispanic White birthing individuals. Rates of in-hospital mortality (4831 Black newborns [15.6%]; 3009 Hispanic newborns [14.9%]; and 4886 White newborns [14.4%]), NEC (2374 Black newborns [7.8%]; 1359 Hispanic newborns [6.9%]; and 2137 White newborns [6.5%]), LOS (3846 Black newborns [13.5%]; 2258 Hispanic newborns [12.3%]; and 3575 White newborns [11.5%]), and sIVH (2919 Black newborns [10.3%]; 1673 Hispanic newborns [9.2%]; and 2800 White newborns [9.1%]) were highest among Black and lowest among White newborns. Chronic lung disease and pneumothorax rates were lowest among Black and highest among White newborns. Over the study period, mortality rate differences were slightly higher for Black than White newborns, with no differences by 2022. NEC and LOS rates were consistently higher among Black than White newborns. By 2022, Black newborns had higher rates of NEC (rate difference, 1.3 percentage points; 95% CI, 0.46-2.2 percentage points) and LOS (rate difference, 2.7 percentage points; 95% CI, 1.4-4.0 percentage points). sIVH rates were higher for Black newborns in some years, whereas severe retinopathy of prematurity rates were lower. Hispanic newborns had mortality and complication rates similar to those of White newborns. Black and Hispanic newborns had lower respiratory complication rates and higher complication-free survival than White newborns.Conclusions and relevanceIn this cohort study, there were no differences in mortality rates between Black and White newborns, but Black newborns had higher rates of NEC and LOS. Continued quality improvement and addressing social determinants of health are critical for promoting health equity in hospital outcomes and beyond.

Dataset Information

An improved method to extract DNA from 1 ml of uncultured amniotic fluid from patients at less than 16 weeks' gestation.

Publications

An improved method to extract DNA from 1 ml of uncultured amniotic fluid from patients at less than 16 weeks' gestation.

Similar Datasets

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