Collection, storage and analysis of non-invasive genetic material in primate biology

2003 ◽  
pp. 295-308 ◽  
Author(s):  
Benoît Goossens ◽  
Nicola Anthony ◽  
Kathryn Jeffery ◽  
Mireille Johnson-Bawe ◽  
Michael W. Bruford
Keyword(s):  
Genetic Material ◽  
Non Invasive

Application of real-time PCR of sex-independent insertion-deletion polymorphisms to determine fetal sex using cell-free fetal DNA from maternal plasma

2015 ◽  
Vol 53 (8) ◽  
Author(s):  
Sherry Sze Yee Ho ◽  
Angela Barrett ◽  
Henna Thadani ◽  
Cecille Laureano Asibal ◽  
Evelyn Siew-Chuan Koay ◽  
...  
Keyword(s):  
Real Time ◽  
Y Chromosome ◽  
Real Time Pcr ◽  
Genetic Material ◽  
Maternal Plasma ◽  
Fetal Sex ◽  
Female Fetus ◽  
Non Invasive ◽  
Fetal Dna ◽  
Cell Free Fetal Dna

AbstractPrenatal diagnosis of sex-linked disorders requires invasive procedures, carrying a risk of miscarriage of up to 1%. Cell-free fetal DNA (cffDNA) present in cell-free DNA (cfDNA) from maternal plasma offers a non-invasive source of fetal genetic material for analysis. Detection of Y-chromosome sequences in cfDNA indicates presence of a male fetus; in the absence of a Y-chromosome signal a female fetus is inferred. We aimed to validate the clinical utility of insertion-deletion polymorphisms (INDELs) to confirm presence of a female fetus using cffDNA.Quantitative real-time PCR (qPCR) for the Y-chromosome-specific sequence,Fetal sex was correctly determined in 77/82 (93.9%) cfDNA samples.We have validated a non-invasive prenatal test to confirm fetal sex as early as 6 gestational weeks using cffDNA from maternal plasma.

scholarly journals Non-invasive preimplantation genetic testing (niPGT): the next revolution in reproductive genetics?

2019 ◽  
Vol 26 (1) ◽  
pp. 16-42 ◽  
Author(s):  
Megan Leaver ◽  
Dagan Wells
Keyword(s):  
Genetic Testing ◽  
Clinical Diagnosis ◽  
Embryo Culture ◽  
Culture Media ◽  
Genetic Material ◽  
Preimplantation Embryos ◽  
Original Research ◽  
Preimplantation Genetic Testing ◽  
Non Invasive ◽  
The Impact

Abstract BACKGROUND Preimplantation genetic testing (PGT) encompasses methods that allow embryos to be tested for severe inherited conditions or for chromosome abnormalities, relevant to embryo health and viability. In order to obtain embryonic genetic material for analysis, a biopsy is required, involving the removal of one or more cells. This invasive procedure greatly increases the costs of PGT and there have been concerns that embryo viability could be compromised in some cases. The recent discovery of DNA within the blastocoele fluid (BF) of blastocysts and in spent embryo culture media (SCM) has led to interest in the development of non-invasive methods of PGT (niPGT). OBJECTIVE AND RATIONALE This review evaluates the current scientific evidence regarding non-invasive genetic assessment of preimplantation embryos. The success of different PGT methodologies in collecting and analysing extra-embryonic DNA is evaluated, and consideration is given to the potential biological and technical hindrances to obtaining a reliable clinical diagnosis. SEARCH METHODS Original research and review papers concerning niPGT were sourced by searching PubMed and Google Scholar databases until July 2019. Searches comprised the keywords: ‘non-invasive’; ‘cell-free DNA’; ‘blastocentesis’; ‘blastocoel fluid’; ‘spent culture media’; ‘embryo culture medium’; ‘preimplantation genetic testing’; ‘preimplantation genetic diagnosis’; ‘preimplantation genetic screening’; and ‘aneuploidy’. OUTCOMES Embryonic DNA is frequently detectable in BF and SCM of embryos produced during IVF treatment. Initial studies have achieved some success when performing cytogenetic and molecular genetic analysis. However, in many cases, the efficiency has been restricted by technical complications associated with the low quantity and quality of the DNA. Reported levels of ploidy agreement between SCM/BF samples and biopsied embryonic cells vary widely. In some cases, a discrepancy with respect to cytogenetic data obtained after trophectoderm biopsy may be attributable to embryonic mosaicism or DNA contamination (usually of maternal origin). Some research indicates that aneuploid cells are preferentially eliminated from the embryo, suggesting that their DNA might be over-represented in SCM and BF samples; this hypothesis requires further investigation. WIDER IMPLICATIONS Available data suggest that BF and SCM samples frequently provide DNA templates suitable for genetic analyses, offering a potential means of PGT that is less expensive than traditional methods, requires less micromanipulation skill and poses a lower risk to embryos. Critically, DNA isolation and amplification protocols must be optimised to reproducibly obtain an accurate clinical diagnosis, whilst minimising the impact of confounding factors such as contamination. Further investigations are required to understand the mechanisms underlying the release of embryonic DNA and to determine the extent to which this material reflects the true genetic status of the corresponding embryo. Currently, the clinic al potential of niPGT remains unknown.

scholarly journals Invasive Prenatal Diagnosis: Chorionic Villus Sampling

2015 ◽  
Vol 9 (3) ◽  
pp. 293-306
Author(s):  
Panos Antsaklis ◽  
Aris Antsaklis ◽  
Michael Sindos ◽  
Fotodotis M Malamas
Keyword(s):  
Prenatal Diagnosis ◽  
Obstet Gynecol ◽  
Genetic Material ◽  
Chorionic Villus ◽  
First Trimester ◽  
Chorionic Villus Sampling ◽  
Maternal Circulation ◽  
Maternal Cell ◽  
Non Invasive ◽  
Limb Reduction

ABSTRACT Chorionic villus sampling (CVS) is the method of choice for first trimester invasive prenatal diagnosis. In expert hands, it is nowadays considered as safe as amniocentesis and has the advantage of an earlier diagnosis. In this review, we describe the technique of the procedure, its indications and contraindications and the requirements concerning adequate training and optimum clinical practice. We also discuss issues concerning the safety of the procedure in singleton and multiple pregnancies, other complications and controversies, such as the association with limb reduction defects and pre-eclampsia, as well as diagnostic problems and dilemmas, such as maternal cell contamination and confined placental mosaicism. We also describe new and promising methods of non-invasive diagnosis, based on the isolation and analysis of fetal cells or cell-free fetal genetic material from the maternal circulation, that aim to replace the invasive methods of prenatal diagnosis in the future. How to cite this article Sindos M, Malamas FM, Antsaklis P, Antsaklis A. Invasive Prenatal Diagnosis: Chorionic Villus Sampling. Donald School J Ultrasound Obstet Gynecol 2015; 9(3):293-306.

scholarly journals The Validation of a Non-Invasive Skin Sampling Device for Detecting Cetacean Poxvirus

Animals ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 2814
Author(s):  
Simone Segura-Göthlin ◽  
Antonio Fernández ◽  
Manuel Arbelo ◽  
Idaira Felipe-Jiménez ◽  
Ana Colom-Rivero ◽  
...  
Keyword(s):  
Preliminary Investigation ◽  
Genetic Material ◽  
Skin Lesions ◽  
Sampling Procedure ◽  
Sampling Device ◽  
Potential Health ◽  
Non Invasive ◽  
Skin Biopsies ◽  
Human Care ◽  
Free Ranging

Poxvirus-like lesions are widely used as a potential health indicator in cetaceans, although for this application, corroboration of Poxvirus skin disease is imperative. Aiming to address skin biopsies intrusiveness, a preliminary investigation of a non-invasive skin sampling procedure to molecularly detect CePV-1 in 12 tattoo-like-lesions from two free-ranging stranded cetaceans in the Canary Islands was performed. Skin lesions were brushed with cytology cell samplers (CCSs) and placed into 1.5 mL microcentrifuge tubes with 1 mL of RNAlaterTM Stabilization Solution. For factual comparisons, DNA extractions from sloughed skin obtained with CCS and biopsies from the same lesions were accomplished with DNA Tissue Kit STM (QuickGene, Kurabo, Japan). Moreover, a second DNA extraction from sloughed skin with DNeasyTM Blood and Tissue Kit (Qiagen, Inc., Valencia, CA, USA) was performed to ascertain kit suitability for CCS. Molecular detection of CePV-1 was performed through a real-time PCR. As a result, a 91.7% and 83.3% rates of positivity were obtained with biopsies and CCS through Quickgene, respectively, compared to the rate of 100% using CCS with Qiagen. Accordingly, CCS is a reliable non-invasive sampling device to obtain sufficient genetic material to be analyzed for CePV-1 in tattoo-skin-lesions as well as for other purposes in cetaceans under human care.

scholarly journals Remote collection of animal DNA and its applications in conservation management and understanding the population biology of rare and cryptic species

2003 ◽  
Vol 30 (1) ◽  
pp. 1 ◽  
Author(s):  
Maxine P. Piggott ◽  
Andrea C. Taylor
Keyword(s):  
Population Biology ◽  
Phylogenetic Analyses ◽  
Genetic Material ◽  
Home Range Size ◽  
Pilot Studies ◽  
Tissue Samples ◽  
Non Invasive ◽  
Genetic Sampling ◽  
Increased Risk ◽  
Recent Developments

Obtaining useful information about elusive or endangered species can be logistically difficult, particularly if relying entirely on field signs such as hair, feathers or faeces. However, recent developments in molecular technology add substantially to the utility of such 'non-invasive' samples, which provide a source of DNA that can be used to identify not only species but also individuals and their gender. This provides great potential to improve the accuracy of abundance estimates and determine behavioural parameters, such as home-range size, individual habitat and dietary preferences, and even some forms of social interaction. Non-invasive samples can also be a useful alternative to blood or tissue samples (the collection of which traditionally has required trapping of animals) as genetic material for applications such as relatedness, population genetic and phylogenetic analyses. Despite the huge potential of non-invasive genetic sampling, the current technology does have limitations. The low quantity and quality of DNA often obtained from such sources results in an increased risk of genotyping errors, which may lead to incorrect inferences, particularly false identification of individuals. Appropriate precautions and pilot studies are required to minimise these risks, and in some cases it may be wise to employ traditional methods when they are adequate.

What Potent Blood: Non-Invasive Prenatal Genetic Diagnosis and the Transformation of Modern Prenatal Care

2007 ◽  
Vol 33 (1) ◽  
pp. 9-53 ◽  
Author(s):  
Carolyn Jacobs Chachkin
Keyword(s):  
Genetic Disorders ◽  
Genetic Disorder ◽  
Genetic Material ◽  
Genetic Diagnosis ◽  
Prenatal Testing ◽  
Maternal Blood ◽  
Non Invasive ◽  
Tay Sachs Disease ◽  
Promising Solution ◽  
Low Levels

What potent blood hath modest May,What fiery force the earth renews,The wealth of forms, the flush of hues ….—Ralph Waldo EmersonSomeday soon, virtually any pregnant woman will be able to learn — with 98-99% accuracy — whether her fetus has contracted a serious genetic disorder by undergoing nothing more than an inexpensive, non-invasive blood test. For years, scientists have sought a method of prenatal testing that could boast both high levels of accuracy and low levels of risk. The most promising solution lies in an exciting recent discovery: tiny quantities of fetal cells and DNA cross over into the mother's bloodstream during pregnancy. If the fetal genetic material can be successfully isolated from the maternal blood, it can be used to diagnose a number of genetic disorders, such as Down Syndrome, cystic fibrosis, Tay-Sachs disease, and sickle cell anemia. Indeed, researchers have spent the last decade developing ways to accomplish this.

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