scholarly journals NIPT Technique Based on the Use of Long Chimeric DNA Reads

Genes ◽  
2020 ◽  
Vol 11 (6) ◽  
pp. 590
Author(s):  
Vera Belova ◽  
Daria Plakhina ◽  
Sergey Evfratov ◽  
Kirill Tsukanov ◽  
Gennady Khvorykh ◽  
...  
Keyword(s):  
Pregnant Women ◽  
Predictive Value ◽  
High Performance ◽  
High Throughput Sequencing ◽  
Prenatal Testing ◽  
First Trimester ◽  
Extracellular Dna ◽  
Trimester Screening ◽  
Sensitivity Specificity ◽  
Analytical Approaches

Non-invasive prenatal testing (NIPT) for aneuploidy on Chromosomes 21 (T21), 18 (T18) and 13 (T13) is actively used in clinical practice around the world. One of the limitations of the wider implementation of this test is the high cost of the analysis itself, as high-throughput sequencing is still relatively expensive. At the same time, there is an increasing trend in the length of reads yielded by sequencers. Since extracellular DNA is short, in the order of 140–160 bp, it is not possible to effectively use long reads. The authors used high-performance sequencing of cell-free DNA (cfDNA) libraries that went through additional stages of enzymatic fragmentation and random ligation of the resulting products to create long chimeric reads. The authors used a controlled set of samples to analyze a set of cfDNA samples from pregnant women with a high risk of fetus aneuploidy according to the results of the first trimester screening and confirmed by invasive karyotyping of the fetus using laboratory and analytical approaches developed by the authors. They evaluated the sensitivity, specificity, PPV (positive predictive value), and NPV (negative predictive value) of the results. The authors developed a technique for constructing long chimeric reads from short cfDNA fragments and validated the test using a control set of extracellular DNA samples obtained from pregnant women. The obtained sensitivity and specificity parameters of the NIPT developed by the authors corresponded to the approaches proposed earlier (99.93% and 99.14% for T21; 100% and 98.34% for T18; 100% and 99.17% for T13, respectively).

scholarly journals NIPT technique based on the use of long chimeric DNA reads

2020 ◽  
Author(s):  
Vera Belova ◽  
Daria Plakhina ◽  
Sergey Evfratov ◽  
Kirill Tsukanov ◽  
Gennady Khvorykh ◽  
...  
Keyword(s):  
Pregnant Women ◽  
High Performance ◽  
High Throughput Sequencing ◽  
Prenatal Testing ◽  
First Trimester ◽  
Extracellular Dna ◽  
Non Invasive ◽  
Trimester Screening ◽  
Sensitivity Specificity ◽  
Analytical Approaches

Abstract Background: Non-invasive prenatal testing for aneuploidy on chromosomes 21, 18 and 13 is actively used in clinical practice around the world. One of the limitations of the wider implementation of this test is the high cost of the analysis itself, as the high throughput sequencing is still relatively expensive. At the same time, there is a trend of increase of the length of reads yielded by sequencers. Since extracellular DNA is short, in the order of 140-160 bp, it is not possible to effectively use long reads.Results: The authors used high-performance sequencing of cfDNA libraries that went through additional stages of enzymatic fragmentation and random ligation of the resulting products to create long chimeric reads. The authors used a controlled set of samples to analyze a set of cfDNA samples of pregnant women with a high risk of fetus aneuploidy according to the results of the first trimester screening and confirmed by invasive karyotyping of the fetus using laboratory and analytical approaches developed by the authors. They evaluated the sensitivity, specificity, PPV and NPV of the results.Conclusions: The authors developed a technique for constructing long chimeric reads from short cfDNA fragments and validated the test using a control set of extracellular DNA samples obtained from pregnant women. The obtained sensitivity and specificity parameters of the NIPT developed by the authors corresponded to the approaches proposed earlier (99, 93% and 99.14% for 21 trisomy; 100% and 98.34% for 18 trisomy; 100% and 99.17% for 13 trisomy, respectively).

Maternal Serum EG-VEGF as A First Trimester Predictor of Hypertensive Disorders of Pregnancy: A Prospective Cohort Study

2021 ◽  
Author(s):  
Shiyu Zeng ◽  
Ling Yu ◽  
Yiling Ding ◽  
Mengyuan Yang
Keyword(s):  
Cohort Study ◽  
Growth Factor ◽  
Prediction Model ◽  
Pregnant Women ◽  
Predictive Value ◽  
Prospective Cohort ◽  
First Trimester ◽  
Hypertensive Disorders Of Pregnancy ◽  
Hypertensive Disorders ◽  
Sensitivity Specificity

Abstract Background This study aims to explore whether plasma endocrine gland-derived vascular endothelial growth factor (EG-VEGF) in the first trimester can be used as a predictor of hypertensive disorders of pregnancy (HDP), and compare it with placental growth factor (PlGF) and soluble fms-like tyrosine kinase-1 (sFlt-1) to evaluate its prediction of HDP value. Methods This is a prospective cohort study that records the medical history of the pregnant women included in the study at 11–13 weeks’ gestation, and analyzes serum biochemical markers including EG-VEGF, PIGF, sFlt-1 and sFlt-1/PIGF. The predictive values of these tests were determined. We used the receiver operating characteristic (ROC) curve to find the optimal cut-off value for each biomarker and compare the operating characteristics (sensitivity, specificity). Logistic regression analysis was used to create a prediction model for HDP based on maternal characteristics and maternal biochemistry. Results Data were obtained from 205 pregnant women. 17 cases were diagnosed with HDP, the incidence rate was 8.2% (17/205). Women who developed HDP had a significantly higher body mass index (BMI) and mean arterial pressure (MAP). Serum EG-VEGF levels in the first trimester are significantly higher in pregnant women with HDP. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value(NPV) of serum EG-VEGF levels more than 227.83 pg/ml for predicting HDP were 43%, 93%, 86% and 62%, respectively. We established a prediction model in the first trimester include maternal BMI, MAP, and EG-VEGF, with an AUC of 0.8861 (95%CI: 0.7905–0.9818), which is better than using EG-VEGF alone (AUC: 0.66). Conclusion This study demonstrated that serum EG-VEGF is a promising biomarker for predicting HDP in the first trimester. It has better predictive performance compared with the currently used biomarkers like PIGF and sFlt-1. Combining maternal clinical characteristics and biochemical tests at 11–13 weeks can effectively identify women at high risk of HDP.

Use of endocervical trophoblasts in noninvasive prenatal testing for common aneuploidies

2020 ◽  
Vol 19 (4) ◽  
pp. 75-80
Author(s):  
E.V. Musatova ◽  
◽  
M.V. Kapustina ◽  
M.E. Minzhenkova ◽  
Zh.G. Markova ◽  
...  
Keyword(s):  
Pregnant Women ◽  
Prenatal Testing ◽  
First Trimester ◽  
Dna Probes ◽  
Fish Analysis ◽  
Trophoblast Cells ◽  
Noninvasive Prenatal Testing ◽  
Chorionic Villi ◽  
Trimester Screening ◽  
Reliable Methods

The article presents an experience of isolation of trophoblast cells from cervical samples of pregnant women and demonstrates a possibility of determining aneuploid cells in cervical samples using FISH analysis. Objective. To study methods of effective isolation and reliable detection of trophoblasts for genetic pathology analysis. Patients and methods. The participants of the study were three pregnant women, who according to the findings of the combined first trimester screening test were referred to the group with a high risk for fetal aneuploidy. The terms of gestation varied from 12 to 14 wks. Immunocytochemical detection of trophoblast cells was performed with FITC-labelled HLA-G antibodies. FISH was performed with the use of locus-specific DNA probes on chromosomes 13/21 and a DNA probe on chromosome 18 centromere. The chromosome set of a fetus was verified by the results of standard cytogenetic analysis of semi-direct chromosome preparations from cytotrophoblast cells obtained by chorionic villi biopsy. Results. HLA-G-positive cells were found in all examined cytological specimens. FISH analysis with the use of DNA probes on chronomomes, whose trisomies are compatible with live birth, detected aneuploid cells in all three cases. Conclusion. The use of cervical trophoblasts for noninvasive collection of information about the genetic status of the developing fetus at present needs further study of effective and reliable methods of their isolation, detection and analysis. Key words: aneuploidy, throphoblast cells, NIPT, prenatal diagnosis, HLA-G

scholarly journals Noninvasive prenatal testing in megacity

2021 ◽  
Vol 2 (2) ◽  
pp. 75-83
Author(s):  
Anton S. Olenev ◽  
Ekaterina N. Songolova ◽  
Alfina A. Yakshibaeva
Keyword(s):  
Prenatal Diagnosis ◽  
Chromosomal Abnormalities ◽  
False Negative ◽  
Prenatal Testing ◽  
First Trimester ◽  
Extracellular Dna ◽  
Prenatal Diagnostics ◽  
Individual Risk ◽  
Negative Results ◽  
Trimester Screening

Introduction. The article presents an analysis of the use of non-invasive prenatal testing for chromosomal abnormalities in the fetal extracellular DNA in the blood of pregnant women in Moscow. Materials and methods. When processing materials and research results, authors considered all available clinical data: findings of an ultrasound examination, medical history and results of additional laboratory tests. Results. The article presents results of invasive prenatal diagnostics and pregnancy outcomes in patients with high NIPT risks. Discussion. Authors analyzed diagnostic capabilities of NIPT, its limitations, false-positive and false- negative results, and described 3 cases of prenatal diagnosis of fetal aneuploidy identified in patients who are in the «gray zone» of risk based on the results of combined screening of the first trimester of pregnancy. Conclusion. Prenatal diagnosis of the described aneuploidy is difficult due to individual risk of developing chromosomal abnormalities of the fetus based on the results of combined first-trimester screening in the range of 1:101-1: 2500 and lack of indications for invasive prenatal diagnostics.

scholarly journals Serum hypoxia-inducible factor-1α and uterine artery Doppler ultrasound during the first trimester for prediction of preeclampsia

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Wasinee Tianthong ◽  
Vorapong Phupong
Keyword(s):  
Pregnant Women ◽  
Doppler Ultrasound ◽  
Predictive Value ◽  
Uterine Artery ◽  
Hypoxia Inducible Factor ◽  
First Trimester ◽  
Singleton Pregnancy ◽  
Uterine Artery Doppler ◽  
Hypoxia Inducible Factor 1Α ◽  
Sensitivity Specificity

AbstractThe objective of this study was to determine the predictive value of serum hypoxia-inducible factor-1α (HIF-1α) combined with uterine artery Doppler in singleton pregnancy during 11–13+6 weeks of gestation for preeclampsia. This prospective observational study was conducted in singleton pregnant women at 11–13+6 weeks of gestation who visited the King Chulalongkorn Memorial Hospital, Faculty of Medicine, Chulalongkorn University for antenatal care between February 2019 and May 2020. Serum HIF-1α levels and uterine artery Doppler ultrasound were performed. Pregnancy outcomes were recorded. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of these tests at the optimal cut-off values were determined to predict preeclampsia. A total of 385 participants were analyzed. Of these, 31 cases had preeclampsia (8.1%), and 6 cases of them had early-onset preeclampsia (1.6%). Preeclamptic women had significantly higher serum HIF-1α levels than normal pregnant women (median 1315.2 pg/ml vs. 699.5 pg/ml, p < 0.001). There was no difference in the mean pulsatility (PI) of the uterine artery. Serum HIF-1α levels were higher than 1.45 multiple of median for the gestational age as a cut-off value for predicting preeclampsia; the sensitivity, specificity, PPV, and NPV were 66.7%, 71.5%, 17.2%, and 96.2%, respectively. When a combination of abnormal serum HIF-1α levels and abnormal uterine artery Doppler PI (above the 95th percentile) were used as a predictive value to predict preeclampsia, the sensitivity, specificity, PPV, and NPV were 74.2%, 67.2%, 16.6%, and 96.8%, respectively. This study showed that the serum HIF-1α levels with or without uterine artery Doppler at 11–13+6 weeks of gestation were effective in predicting preeclampsia.

scholarly journals Efficiency of non-invasive prenatal screening in pregnant women at advanced maternal age

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Hui Zhu ◽  
Xiaoxiao Jin ◽  
Yuqing Xu ◽  
Weihua Zhang ◽  
Xiaodan Liu ◽  
...  
Keyword(s):  
High Risk ◽  
Pregnant Women ◽  
Predictive Value ◽  
Trisomy 21 ◽  
Maternal Age ◽  
Prenatal Screening ◽  
Advanced Maternal Age ◽  
Youden Index ◽  
Non Invasive ◽  
Sensitivity Specificity

Abstract Background Non-invasive prenatal screening (NIPS) is widely used as the alternative choice for pregnant women at high-risk of fetal aneuploidy. However, whether NIPS has a good detective efficiency for pregnant women at advanced maternal age (AMA) has not been fully studied especially in Chinese women. Methods Twenty-nine thousand three hundred forty-three pregnant women at AMA with singleton pregnancy who received NIPS and followed-up were recruited. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV), receiver operating characteristic (ROC) curves and the Youden Index for detecting fetal chromosomal aneuploidies were analyzed. The relationship between maternal age and common fetal chromosomal aneuploidy was observed. Results The sensitivity, specificity, PPV, NPV of NIPS for detecting fetal trisomy 21 were 99.11, 99.96, 90.98, and 100%, respectively. These same parameters for detecting fetal trisomy 18 were 100, 99.94, 67.92, and 100%, respectively. Finally, these parameters for detecting trisomy 13 were 100, 99.96, 27.78, and 100%, respectively. The prevalence of fetal trisomy 21 increased exponentially with maternal age. The high-risk percentage incidence rate of fetal trisomy 21 was significantly higher in the pregnant women at 37 years old or above than that in pregnant women at 35 to 37 years old. (Youden index = 37). Conclusion It is indicated that NIPS is an effective prenatal screening method for pregnant women at AMA.

scholarly journals Radioimmunoassay of plasma ouabain in healthy and pregnant individuals

2000 ◽  
Vol 165 (3) ◽  
pp. 669-677 ◽  
Author(s):  
O Vakkuri ◽  
SS Arnason ◽  
A Pouta ◽  
O Vuolteenaho ◽  
J Leppaluoto
Keyword(s):  
Pregnant Women ◽  
Human Plasma ◽  
High Performance ◽  
Chemical Nature ◽  
Solid Phase ◽  
Plasma Concentrations ◽  
First Trimester ◽  
Plasma Samples ◽  
Third Trimester ◽  
Endogenous Substances

Ouabain was recently isolated from human plasma, bovine hypothalamus and bovine adrenal in attempts to identify endogenous substances inhibiting the cell membrane sodium pump. A number of radioimmunoassays have been developed in order to study the clinical significance of ouabain. The results have been controversial with regard to the presence and chemical nature of plasma ouabain-like immunoreactivity. We have now measured ouabain in healthy and pregnant individuals using solid-phase extraction of plasma samples followed by a new radioimmunoassay with the extraordinary sensitivity of at least 2 fmol/tube (5 pmol/l). Plasma extracts, a previously isolated human plasma ouabain-like compound and bovine hypothalamic inhibitory factor displaced the tracer in parallel and eluted identically with ouabain in high-performance liquid chromatography. Plasma ouabain immunoreactivity was found to be much lower than reported previously: 12.6+/-1.3 pmol/l in healthy men (mean+/-s.e., n=20) and 9.4+/-0.7 pmol/l in women (n=14). In pregnant women (n=28) plasma ouabain concentration was 16.3+/-4.0 pmol/l during the first trimester, 18.8+/-4.3 pmol/l during the second trimester and 24.3+/-4.0 pmol/l during the third trimester (all P<0.01 compared with non-pregnant women). Plasma ouabain 3-5 days after the delivery was 13.6+/-1.1 pmol/l (n=10, P<0.05-0.01 compared with second and third trimesters). The pregnancy-related changes in the plasma concentrations of ouabain resembled those of cortisol. Therefore cortisol was measured from the same plasma samples and a significant positive correlation was found (r=0.512, P=0.006). The similar profiles of plasma ouabain and cortisol during pregnancy and their rapid decreases postpartum are consistent with the adrenal cortical origin of ouabain and also show that the secretions of these hormones are possibly under the control of same factors.

scholarly journals The Effect of Down Syndrome Biochemical Markers on Predicting Severity of Preeclampsia

2020 ◽  
Author(s):  
AYSE OZBAN
Keyword(s):  
Down Syndrome ◽  
Pregnant Women ◽  
Biochemical Markers ◽  
First Trimester ◽  
Maternal Serum ◽  
Screening Tests ◽  
Second Trimester ◽  
Study Results ◽  
Second Trimester Screening ◽  
Trimester Screening

Abstract Objective: This study aims to determine whether it is possible to predict preeclampsia by comparing postpartum results and test results of the pregnant women diagnosed with preeclampsia, whose first and/or second trimester screening tests were accessible, and to demonstrate the predictability of severity and week of onset.Background: 204 patients underwent renal transplantation in our center and 84 of them were female. Five of our patients (one of them had two births) gave birth to a total of 6 pregnancies.Method: 135 patients were diagnosed with preeclampsia and their first and/or second trimester screening tests were accessible, and 366 control participants gave birth to a healthy baby between 37-41 weeks after standard follow-up period for pregnancy and their screening tests were also accessible.Results: The study results show that the first trimester maternal serum PAPP-A level is significantly low in preeclamptic pregnant women, and that the second trimester maternal serum AFP and hCG levels are significantly high and uE3 levels are significantly low The results also suggest that the first and second trimester Down syndrome biochemical markers can be used in preeclampsia screening.Conclusion: Among these markers, uE3 is the parameter which affects the possibility of preeclampsia the most. However, the first and second trimester Down syndrome biochemical markers are not effective in predicting the severity and onset week of preeclampsia.

scholarly journals Prediction of preeclampsia using a combination of serum micro RNA-210 and uterine artery Doppler ultrasound

2021 ◽  
Vol 104 (3) ◽  
pp. 003685042110368
Author(s):  
Ananya Trongpisutsak ◽  
Vorapong Phupong
Keyword(s):  
Pregnant Women ◽  
Doppler Ultrasound ◽  
Predictive Value ◽  
Early Onset ◽  
Uterine Artery ◽  
Micro Rna ◽  
Predictive Values ◽  
Uterine Artery Doppler ◽  
Early Onset Preeclampsia ◽  
Sensitivity Specificity

The objective was to determine whether a combination of serum micro RNA-210 level and uterine artery Doppler can predict preeclampsia in pregnant women at 16–24 weeks gestation. A prospective observational study conducted in singleton pregnant women at 16–24 weeks of gestation who had prenatal care at the King Chulalongkorn Memorial Hospital, Department of Obstetrics and Gynecology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand between 2017 and 2018. Uterine artery Doppler ultrasound and blood testing for serum micro RNA-210 were performed. Pregnancy outcomes were recorded. Optimal cut-off for uterine artery pulsatility index (PI) and serum micro RNA-210 were obtained to calculate the predictive values for preeclampsia. Data from 443 participants were analyzed. Twenty-two cases developed preeclampsia (5.0%) and seven of these preeclamptic cases had early-onset preeclampsia (1.6%). Pregnant women with preeclampsia had higher mean PI of the uterine artery (1.34 ± 0.52 vs 0.98 ± 0.28, p = 0.004), higher detection rates of diastolic notching (45.5% vs 11.2%, p < 0.001), and lower median serum micro RNA-210 level (22.86 vs 795.78, p < 0.001) than pregnant women without preeclampsia. Using abnormal serum micro RNA-210 level, abnormal mean PI or uterine artery diastolic notches to predict for preeclampsia, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were 95.5%, 54.9%, 10.0%, and 99.6%, respectively. For early-onset preeclampsia prediction, the sensitivity, specificity, PPV, and NPV were 100.0%, 53.2%, 3.3%, and 100.0%, respectively. This study demonstrated that a combination of serum micro RNA-210 and uterine artery Doppler is effective in predicting preeclampsia in the second trimester.

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