Invented by Charles R. Cantor, Grace DeSantis, Reinhold Mueller, Mathias Ehrich, Sequenom Inc

The market for methods for non-invasive evaluation of genetic variation is rapidly expanding as advancements in technology and research continue to revolutionize the field of genetics. Non-invasive methods refer to techniques that do not require invasive procedures such as biopsies or blood samples, making them more convenient and less invasive for patients. Genetic variation refers to differences in DNA sequences among individuals, which can have significant implications for disease susceptibility, drug response, and overall health. Traditional methods of evaluating genetic variation often involve invasive procedures, which can be costly, time-consuming, and uncomfortable for patients. However, with the advent of non-invasive methods, the process has become more accessible and efficient. One of the most well-known non-invasive methods for evaluating genetic variation is through the use of saliva or buccal swabs. These samples can be easily collected by individuals themselves or healthcare professionals and sent to a laboratory for analysis. This method is particularly useful for studying genetic variation in large populations or for screening purposes. Another non-invasive method gaining popularity is the use of urine samples. Urine contains cell-free DNA, which can be extracted and analyzed to identify genetic variations. This method is particularly useful for monitoring genetic changes over time and for detecting genetic abnormalities in prenatal screening. Non-invasive prenatal testing (NIPT) is another area where the market for genetic variation evaluation is growing rapidly. NIPT involves analyzing cell-free DNA from the mother’s blood to detect chromosomal abnormalities in the fetus. This method has significantly improved the accuracy and safety of prenatal screening, reducing the need for invasive procedures such as amniocentesis. Advancements in technology, such as next-generation sequencing (NGS), have also contributed to the growth of the market for non-invasive evaluation of genetic variation. NGS allows for the simultaneous analysis of multiple genes or even the entire genome, providing a comprehensive view of an individual’s genetic makeup. This technology has made it possible to identify rare genetic variants and has opened up new avenues for personalized medicine. The market for non-invasive evaluation of genetic variation is not limited to healthcare settings. It also extends to areas such as forensics, where DNA analysis plays a crucial role in identifying individuals and solving crimes. Non-invasive methods, such as analyzing DNA from hair, skin cells, or even touch DNA, have made it easier to collect and analyze genetic evidence in forensic investigations. As the demand for personalized medicine and genetic testing continues to rise, the market for non-invasive evaluation of genetic variation is expected to grow even further. The convenience, accuracy, and cost-effectiveness of non-invasive methods make them an attractive option for both healthcare providers and individuals seeking genetic information. Additionally, ongoing research and technological advancements are likely to lead to the development of new and improved non-invasive methods, further expanding the market. In conclusion, the market for methods for non-invasive evaluation of genetic variation is expanding rapidly due to advancements in technology, increased demand for personalized medicine, and the convenience and accuracy of non-invasive methods. These methods have revolutionized the field of genetics by making genetic testing more accessible, efficient, and less invasive for patients. As research and technology continue to advance, the market is expected to grow even further, opening up new possibilities for personalized medicine and genetic research.

The Sequenom Inc invention works as follows

The technology provided herein is a part of methods, processes, and apparatuses that are used for the non-invasive evaluation of genetic variation.

Background for Methods for non-invasive evaluation of genetic variation

The genetic information of living organisms, such as animals, plants, and microorganisms, and other forms that replicate genetic information, like viruses, is encoded by deoxyribonucleic or ribonucleic acids. Genetic information is represented by a series of nucleotides, or modified nucleotides that represent the primary structure or hypothetical nucleic acid. The human genome is composed of approximately 30,000 genes, which are located on 24 chromosomes. (See The Human Genome by T. Strachan. BIOS Scientific Publishers 1992). Each gene encodes an individual protein that, after transcription and translation, performs a biochemical function in a cell.

Many medical conditions are caused either by genetic variations or other factors. Some genetic variations can cause medical conditions such as hemophilia (blood clotting disorder), thalassemia (thalassemia), Duchenne Muscular Dystrophy, Huntington’s Disease, Alzheimer’s Disease, and Cystic Fibrosis. (Human Genome Mutations by D. N. Cooper & M. Krawczak BIOS Publishers 1993). These genetic diseases are caused by the addition, substitution or deletion of one nucleotide from DNA. Some birth defects can be caused by chromosomal abnormalities, or aneuploidies, including Trisomy 21 and Down’s Syndrome, Trisomy 13 and Patau Syndrome, Trisomy 18 and Edward’s Syndrome, Monosomy X, Turner’s Syndrome, as well as certain sex chromosome anomalies, like Klinefelter’s Syndrome. The sex of the fetus can be determined by sex chromosomes. Some genetic variations can predispose or cause a variety of diseases, including diabetes, arteriosclerosis and obesity.

Identifying genetic variations and variances may lead to the diagnosis or determination of predisposition for a specific medical condition. A genetic variation can facilitate a medical decision or allow for a useful medical procedure. The analysis of cell-free dna can be used to identify one or more variations.

Cell-free DNA is a collection of DNA fragments that are derived from the death of cells and circulate through peripheral blood. High levels of CF DNA can indicate certain clinical conditions, such as cancers, myocardial ischemia, strokes, sepsis and infection. Cell-free fetal (CFF) DNA can also be detected in maternal blood and used to perform noninvasive prenatal diagnosis.

The presence of fetal DNA in maternal plasma can be used to make a non-invasive diagnosis of prenatal pregnancy by analyzing a sample of maternal blood. Quantitative abnormalities in maternal DNA can be linked to a variety of pregnancy-related disorders. These include preeclampsia and preterm labor. They may also indicate antepartum bleeding, invasive placement, fetal Down Syndrome, or other chromosomal anomalies. The analysis of maternal plasma for fetal DNA can provide a valuable tool to monitor the fetomaternal health.

Early detection of pregnancy-related problems, such as complications during pregnancy or genetic defects in the fetus, is crucial, because it allows for early medical intervention, which is necessary to ensure the safety of both mother and fetus. Cells isolated from the fetus are used for prenatal diagnosis through procedures like chorionic villus sampling (CVS), or amniocentesis. These conventional methods, however, are invasive and pose a significant risk to the mother as well as the fetus. According to the National Health Service, a miscarriage rates of 1 to 2 percent are reported following invasive tests such as amniocentesis or chorionic villus sampling (CVS). “An alternative to invasive methods is the use of non-invasive techniques that use circulating CFF DNA.

In some aspects, methods are provided for enriching fetal nucleic acids in sample nucleic acids that include fetal nucleic acids and maternal nucleic acids. The method comprises: (a), obtaining vesicle free nucleics acid from a sample of a pregnant woman, and (b) isolating some or substantially all the vesicle free nucleics acid from the sample resulting in a separation product that is enriched In some embodiments the method also includes (c) analysing nucleic acids in the separation product.

The method also includes analyzing the nucleic acids in a product that is prepared using a process which comprises: (a), obtaining a cell-free circulating nucleic sample from a biological specimen from a female pregnant, which sample contains vesicular and fetal nascent nucleic acids, and (b), separating some or all of the vesicular nascent nucleic from the sample nascent nucleic, and thereby creating a

In some aspects, methods are also provided for enriching vesicle free nucleic acids in sample nucleic acids, including: (a), obtaining cell-free, circulating sample containing vesicle free nucleic and vesicular acid; (b) separating a portion or substantially all the vesicular acid from the sample resulting in a separation, where the vesicle free nucleic in the separation is enriched in comparison to the ves In some embodiments the method also includes (c) analysing nucleic acids in the separation product.

The method also includes analyzing the nucleic acids in a product that is prepared using a process which comprises: (a), obtaining vesicle free nucleic and vesicular acid from a sample of biological material; (b), separating some or all the vesicular acid from the sample and producing a product where the vesicle free nucleic in the product is enriched in comparison to the vesicle free nucleic in the sample.

The method of enriching fetal vesicular nccleic acids in sample nucleic acids that include both fetal and maternal nucleic acids includes (a) obtaining a cell-free circulating nucleic sample from a sample taken from a female pregnant, the sample nucleic sample containing maternal-derived and fetal vesicular nccleic nucleics; (b) separating the maternal-derived and fetal vesicular

The method of enriching fetal nascicular acid in a sample nucleic acids that contains both fetal and maternal nucleic acids includes (a) obtaining a cell-free circulating nucleic sample from a biological specimen from a female pregnant, the sample nucleic sample comprising vesicle free nucleic and vesicular nascicular acid; (b) seperating some or substantially all the vesicular

In some embodiments, the separation of some or substantially all the maternal-derived nucleic acids from the fetal nucleic acids involves contacting the sample with an agent which specifically binds the maternal-derived nucleic acids. In some embodiments separating some or substantial all of the fetal derived vesicular acid from the maternal derived vesicular acid involves contacting the sample with an agent which specifically binds fetal derived vesicular acid.

In some embodiments, the separation of some or substantially of the vesicular nascicular acid from the nucleic sample acid includes centrifuging and ultracentrifugation. In some embodiments separating the vesicular nasciular acid from the nucleic sample acid includes contacting the nucleic sample acid with an agent which specifically binds vesicles containing the vesicular nascicular acid. In some embodiments the agent is an antigen. In certain embodiments, the drug binds specifically to hemopoietic vesicles. In certain embodiments, the drug binds specifically to red blood cell vesicles. In certain instances, the agent binds specifically to CD235a. In certain embodiments, the agent binds specifically to leukocyte vesicles. In certain instances, the agent binds specifically to CD45. In certain embodiments, the agents binds specifically to lymphocyte vesicles. In certain instances, the agent binds specifically to a component of vesicular cells selected from CD4,CD8 or CD20. In certain embodiments, the agents binds specifically to granulocyte vesicles. In certain instances, the drug binds specifically to CD66b. In certain embodiments, the agent binds specifically to vesicles of monocytes. In certain instances, the drug binds specifically to CD14. In certain embodiments, the agents binds specifically to platelet vesicles. In certain instances, the agent binds specifically to a component of vesicular cells selected from CD31. CD41. CD41a. CD42a. CD42b. CD61. and CD62P. In certain embodiments, the agents binds specifically to endothelial cell vesicles. In certain instances, the agent binds specifically to a component of vesicular cells selected from CD31. CD34. CD54. CD62E. CD51. CD105. CD106. CD144. and CD146.

In some embodiments, the process of generating the separation products includes separating components bound to the agent from the sample nucleic acids. In some embodiments separating some or substantially the entire vesicular nascllic acid from the nucleic sample acid also comprises contacting it with an agent which specifically binds a histone that is associated with the nucleic sample acid. In some cases, the agent binds specifically to histone H3.3. In certain instances, the agent binds specifically to histone H1. Histone H1 can be unmethylated in some cases.

In some embodiments the vesicular DNA is contained within a vesicle with a diameter less than 1 micrometer. The diameter can range from 10 nanometers up to 600 nanometers in some cases. The diameter can range from 40 nanometers up to 100 nanometers in some cases.

In some aspects, methods are also provided for enriching fetal nasclic acid in sample nasclic acid that contains fetal nasclic acid and maternal niclic acid. These include: (a), obtaining cell-free, circulating sample RNA from a biological specimen from a female pregnant, which sample RNA comprises a histone associated nucleic species and a histone associated nucleic species; (b), separating all or part of the histone species in the first In some embodiments the method also includes (c) analysing nucleic acids in the separation product.

The patent also provides methods for analyzing the nucleic acids in a product that is prepared using a process which includes: (a), obtaining circulating cell-free sample nucleic acids from a biological specimen from a female pregnant, which sample contains a first histone associated nucleic species, a secondary histone associated nucleic species, maternal nucleic and fetal acid; (b), separating some or substantially the entire first histone related nucleic species from the sample, thus generating a

In some aspects, methods are also provided for enriching nucleic acids with histones in a sample nucleic sample. These include: (a), obtaining a cell-free, circulating sample of nucleic sample from a biological specimen, where the sample nucleic sample contains a first nucleic species associated with histones and a second nucleic species associated with histones; (b), separating all or part of the first nucleic species associated to histones from the sample In some embodiments the method also includes (c) analysing nucleic acids in the separation product.

Also, provided in certain aspects are methods that include analyzing nucleic acids in a product separated by a procedure comprising (a) obtaining a cell-free circulating nucleic sample from a biological specimen, where the sample nucleic sample comprises a histone associated nucleic species and a histone associated nucleic species, and (b), separating all or part of the histone associated nucleic species from the sample sample nucleic, thus generating a product enriched in

The patent also provides methods to enrich fetal DNA in sample nucleic acids that include fetal and maternal nucleic acids. These methods consist of (a) obtaining circulating cell-free sample nucleic from a biological specimen from a female pregnant, the sample nucleic containing a first and second histone associated nucleic species. (b) separating all or part of the second and first histone associated nucleic species in order to produce a separation product that is enriched in the second

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