P12 Direct counting of chromatids in polar bodies with digital PCR – a new method for aneuploidy screening in oocytes

2012 ◽  
Vol 24 ◽  
pp. S50
Author(s):  
A. Daser ◽  
E. Day ◽  
B. Konfortov ◽  
H. Turley ◽  
A. Immesberger ◽  
...  
Keyword(s):  
Digital Pcr ◽  
New Method ◽  
Aneuploidy Screening ◽  
Polar Bodies ◽  
Direct Counting

scholarly journals Preimplantation Genetic Diagnosis—An Overview

2005 ◽  
Vol 53 (3) ◽  
pp. 255-260 ◽  
Author(s):  
Caroline Mackie Ogilvie ◽  
Peter R. Braude ◽  
Paul N. Scriven
Keyword(s):  
Preimplantation Genetic Diagnosis ◽  
Genetic Diagnosis ◽  
Sex Selection ◽  
Hla Typing ◽  
Success Rates ◽  
Aneuploidy Screening ◽  
Reciprocal Translocations ◽  
Public Controversy ◽  
Polar Bodies ◽  
Monogenic Diseases

Since the early 1990s, preimplantation genetic diagnosis (PGD) has been expanding in scope and applications. Selection of female embryos to avoid X-linked disease was carried out first by polymerase chain reaction, then by fluorescence in situ hybridization (FISH), and an ever-increasing number of tests for monogenic diseases have been developed. Couples with chromosome rearrangements such as Robertsonian and reciprocal translocations form a large referral group for most PGD centers and present a special challenge, due to the large number of genetically unbalanced embryos generated by meiotic segregation. Early protocols used blastomeres biopsied from cleavage-stage embryos; testing of first and second polar bodies is now a routine alternative, and blastocyst biopsy can also be used. More recently, the technology has been harnessed to provide PGD-AS, or aneuploidy screening. FISH probes specific for chromosomes commonly found to be aneuploid in early pregnancy loss are used to test blastomeres for aneuploidy, with the aim of replacing euploid embryos and increasing pregnancy rates in groups of women who have poor IVF success rates. More recent application of PGD to areas such as HLA typing and social sex selection have stoked public controversy and concern, while provoking interesting ethical debates and keeping PGD firmly in the public eye.

scholarly journals New Advances of Preimplantation and Prenatal Genetic Screening and Noninvasive Testing as a Potential Predictor of Health Status of Babies

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Tanya Milachich
Keyword(s):  
Genetic Diagnosis ◽  
Noninvasive Testing ◽  
Human Embryos ◽  
Embryonic Cells ◽  
Aneuploidy Screening ◽  
Vitro Fertilization ◽  
Prenatal Genetic Diagnosis ◽  
Polar Bodies ◽  
Comprehensive Chromosome Screening

The current morphologically based selection of human embryos for transfer cannot detect chromosome aneuploidies. So far, only biopsy techniques have been able to screen for chromosomal aneuploidies in the in vitro fertilization (IVF) embryos. Preimplantation genetic diagnosis (PGD) or screening (PGS) involves the biopsy of oocyte polar bodies or embryonic cells and has become a routine clinical procedure in many IVF clinics worldwide, including recent development of comprehensive chromosome screening of all 23 pairs of chromosomes by microarrays for aneuploidy screening. The routine preimplantation and prenatal genetic diagnosis (PND) require testing in an aggressive manner. These procedures may be invasive to the growing embryo and fetus and potentially could compromise the clinical outcome. Therefore the aim of this review is to summarize not only the new knowledge on preimplantation and prenatal genetic diagnosis in humans, but also on the development of potential noninvasive embryo and fetal testing that might play an important role in the future.

scholarly journals Clinical Validation of Novel Chip-Based Digital PCR Platform for Fetal Aneuploidies Screening

Diagnostics ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1131
Author(s):  
Anna Nykel ◽  
Rafał Woźniak ◽  
Agnieszka Gach
Keyword(s):  
Sensitivity And Specificity ◽  
Clinical Performance ◽  
High Sensitivity ◽  
Digital Pcr ◽  
Trisomy 13 ◽  
Prenatal Diagnostics ◽  
Aneuploidy Screening ◽  
Clinical Sensitivity ◽  
Fetal Aneuploidy ◽  
Increased Risk

Fetal aneuploidy is routinely diagnosed by karyotyping. The development of techniques for rapid aneuploidy detection based on the amplification reaction allows cheaper and rapid diagnosis. However, the currently available solutions have limitations. We tested a novel approach as a diagnostic tool in clinical practice. The objective of this study was to provide a clinical performance of the sensitivity and specificity of a novel chip-based digital PCR approach for fetal aneuploidy screening. The study was conducted in 505 pregnant women with increased risk for fetal aneuploidy undergoing invasive prenatal diagnostics. DNA extracted from amniotic fluid or CVS was analyzed for the copy number of chromosomes 13, 18, 21, X, and Y using a new chip-based solution. Performance was assessed by comparing results with findings from karyotyping. Aneuploidy was confirmed in 65/505 cases positive for trisomy 21, 30/505 cases positive for trisomy 18, 14/505 cases positive for trisomy 13 and 21/505 with SCAs. Moreover, 2 cases with triploidy and 2 cases with confirmed mosaicisms of 21 and X chromosomes were detected. Clinical sensitivity and specificity within this study was determined at 100% for T21 (95% CI, 99.26–100%), T18 (95% CI, 99.26–100%), and T13 (95% CI, 99.26–100%). Chip-based digital PCR provides equally high sensitivity and specificity in rapid aneuploidy screening and can be implemented into routine prenatal diagnostics.

scholarly journals Multiple aneuploidies in the oocytes of balanced translocation carriers: a preimplantation genetic diagnosis study using first polar body

Reproduction ◽  
2003 ◽  
pp. 701-711 ◽  
Author(s):  
A Pujol ◽  
M Durban ◽  
J Benet ◽  
I Boiso ◽  
JM Calafell ◽  
...  
Keyword(s):  
Preimplantation Genetic Diagnosis ◽  
Polar Body ◽  
Genetic Diagnosis ◽  
Single Embryo Transfer ◽  
Balanced Translocation ◽  
Aneuploidy Screening ◽  
Robertsonian Translocations ◽  
Polar Bodies ◽  
First Polar Body ◽  
Made In

Preimplantation genetic diagnosis (PGD) of first polar bodies (1PBs) has been used in carriers of balanced chromosomal reorganizations and also for aneuploidy screening. Although an acceptable number of normal or balanced embryos is usually obtained using PGD in translocation carriers, the pregnancy rate is disappointingly low. To determine whether aneuploidy of chromosomes not involved in the chromosome rearrangements could be the cause of the low pregnancy rates achieved, the present authors analysed the segregation products of three translocation carriers, t(8;13)(q24.1;q22) and two Robertsonian (Rob)(13;14), using 1PBs, and afterwards another eight chromosomes in the same 1PBs, for a total of 10 chromosomes in each 1PB, that is chromosomes 1, 8, 13, 14, 15, 16, 17, 18, 21, 22 and X. In the reciprocal translocation, chromosomes with different chromatids due to meiotic recombination were found. Only one out of nine 1PBs was normal for the reorganization products but no aneuploidies were found after PGD in this case. In the two balanced Rob(13;14), six out of 12 and four out of 11 1PBs were normal or balanced for the reorganization but only one oocyte was euploid for all the chromosomes analysed in each case; a single embryo transfer was made in both but no pregnancy was achieved. The incidence of aneuploidy for the chromosomes not involved in the Robertsonian translocations was extremely high (91.7% and 81.8%). Extra chromosomes were present in most of the aneuploid oocytes (81.8% and 90%). The reason for this increase could be the tendency to non-disjunction related to advanced maternal age combined with an interchromosomal effect resulting in the presence of synaptic errors in other chromosome pairs.

scholarly journals Digital PCR: A Sensitive and Precise Method for KIT D816V Quantification in Mastocytosis

2018 ◽  
Vol 64 (3) ◽  
pp. 547-555 ◽  
Author(s):  
Georg Greiner ◽  
Michael Gurbisz ◽  
Franz Ratzinger ◽  
Nadine Witzeneder ◽  
Ingrid Simonitsch-Klupp ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Limit Of Detection ◽  
Systemic Mastocytosis ◽  
Prognostic Significance ◽  
Melting Curve ◽  
Digital Pcr ◽  
New Method ◽  
Sensitive Detection ◽  
Melting Curve Analysis ◽  
Kit D816v

Abstract BACKGROUND The analytically sensitive detection of KIT D816V in blood and bone marrow is important for diagnosing systemic mastocytosis (SM). Additionally, precise quantification of the KIT D816V variant allele fraction (VAF) is relevant clinically because it helps to predict multilineage involvement and prognosis in cases of advanced SM. Digital PCR (dPCR) is a promising new method for sensitive detection and accurate quantification of somatic mutations. METHODS We performed a validation study of dPCR for KIT D816V on 302 peripheral blood and bone marrow samples from 156 patients with mastocytosis for comparison with melting curve analysis after peptide nucleic acid-mediated PCR clamping (clamp-PCR) and allele-specific quantitative real-time PCR (qPCR). RESULTS dPCR showed a limit of detection of 0.01% VAF with a mean CV of 8.5% and identified the mutation in 90% of patients compared with 70% for clamp-PCR (P < 0.001). Moreover, dPCR for KIT D816V was highly concordant with qPCR without systematic deviation of results, and confirmed the clinical value of KIT D816V VAF measurements. Thus, patients with advanced SM showed a significantly higher KIT D816V VAF (median, 2.43%) compared with patients with indolent SM (median, 0.14%; P < 0.001). Moreover, dPCR confirmed the prognostic significance of a high KIT D816V VAF regarding survival (P < 0.001). CONCLUSIONS dPCR for KIT D816V provides a high degree of precision and sensitivity combined with the potential for interlaboratory standardization, which is crucial for the implementation of KIT D816V allele burden measurement. Thus, dPCR is suitable as a new method for KIT D816V testing in patients with mastocytosis.

scholarly journals PENANGGALAN 14C UNTUK MENENTUKAN UMUR PELAPUKAN TANAH DENGAN METODE RADIOKARBON

2012 ◽  
Vol 32 (2) ◽  
pp. 125-134
Author(s):  
Darwin A. Siregar ◽  
Satrio Satrio
Keyword(s):  
Radiocarbon Dating ◽  
Age Determination ◽  
New Method ◽  
Co2 Absorption ◽  
Isotope Hydrology ◽  
Absorption Method ◽  
Carbon Sample ◽  
Material Cost ◽  
Direct Counting

Radiocarbon dating is a tool for age determination of a carbon sample. During the time, synthesis benzene method is used for age determination mentioned. By this method it could be analyzed one sample per day only with high material cost. Lately, it has been developed a new method, namely CO2 absorption method. The latest method is often called as direct counting CO2, because radioisotope of 14C in CO2 is counted directly and converted to age. The aim of the development and the use of the method are supporting some research in isotope hydrology, oceanography, climatology, geology, and archeology by faster, economic and practice. The result of 14C analyses for the same sample using the CO2 absorption when compared to the synthesis benzene method is relatively equal.  

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