COVARIS, INC. — Department of Health and Human Services SBIR Phase I: 172

Description

Summary Sequencing of human DNA has become relatively inexpensive in recent years The DNA sequence the order in which the chemical constituents of the DNA are attached to one another in the DNA molecule holds important information about susceptibility to disease as well as possible usefulness of various therapies for diseases Also modifications to DNA such as methylation where a chemical group is added to the DNA impact health because they can suppress or promote the way genes are used to create proteins For sequencing an entire genome of an individual or understanding the methylation of the sequence a significant amount of DNA that is chemically intact is required A good way to get this DNA is by extracting it from the white blood cells of whole blood But whole blood is difficult to collect and store for later analysis Other fluids such as saliva or cheek cells obtained using a swab have variable amounts of DNA that also might be broken For several years blood spots have been dried on filter paper This allows storage of the DNA until it needs to be used But the application of blood to spots can be variable especially if people do it on their own at home And the extraction of DNA using current methods is not that efficient so that getting enough DNA for analysis is a problem This project aims to create a system for easy collection and storage of blood with high yields of good quality DNA We will do this in two steps First blood will be collected into a special device designed for easy application of the blood and which can dry the blood rapidly in a closed container The container can be handled immediately and shipped even before drying is complete In the second step the part of the device containing the dry blood will be subject to focused ultrasound This has been proven to extract larger quantities of DNA from devices such as blood spot paper than the usual chemical methods employed by doctors and researchers This system thus improves three steps in the blood analysis process Blood collection is made easier by providing a small surface on which the blood is placed drying is made easier because the user does not need to worry about how long to dry or maintaining the drying container in an upright orientation or a special environment shipping and storage is made easier because the samples can be shipped immediately or stored indefinitely in the same container and extraction of the DNA is made more efficient using ultrasound Additionally ultrasound has been proven to break the DNA up into the desired lengths that sequencing machines work with best This project will develop new blood collection drying and DNA extraction systems appropriate for sequencing applications An easy to use collection and drying consumable will have improved ease of use than existing methods such as saliva swabs and dried blood spots and the use of focused ultrasound for extraction will provide greater DNA yields than these methods A validation of a manufacturable consumable coupled with a commercial focused ultrasonicator will be performed by comparing DNA yields from fresh whole blood and dried extracted blood using patient samples