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Diagnostic testing for certain genetic and infectious diseases typically requires that a sample of patient blood, urine, or other tissue be tested for the presence of specific nucleic acid sequences. These tests may be performed in vitro using extracted nucleic acid samples (e.g., blood cells) or in vivo using testing performed on non-extracted cellular samples. Non-extracted testing may be performed on the whole blood sample (e.g., the presence of antigens and antibodies), or on a separated cell fraction of the blood sample (e.g., T-cells). Certain diseases, such as the transfusion-transmitted viral infections (e.g., HIV, HBV, HCV, syphilis, and malaria), and certain genetic disorders (e.g., sickle cell anemia and hemophilia) are routinely diagnosed by testing extracted nucleic acid samples for the presence of specific nucleic acid sequences. The nucleic acid sequences may be the nucleic acid sequences of the virus or genetic disorder itself, or the nucleic acid sequences of the viral strains or genetic variants associated with the disease. Testing of nucleic acid samples extracted from the cellular fraction of a blood sample has been shown to be highly sensitive and specific for certain diseases, and may be more sensitive than antigen and antibody testing.
The sensitivity of nucleic acid testing is important because it directly correlates with the detection of the disease in the patient. Thus, it is generally desirable to perform sample extraction and analysis as efficiently as possible. Unfortunately, current nucleic acid extraction techniques
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.FEBRUARY (20171201.JPG)The invention relates to a method for treating a substrate comprising immobilising at least a biomolecule on at least a part of the substrate, detecting free, i.e. unbound and unbound as a result of detection, biomolecule, and optionally delivering the biologically active biomolecule.
Said substrates may be test samples.
The invention also relates to a biochip comprising a support, at least a biomolecule and a means of detecting a free, i.e. unbound biomolecule present on the substrate.
So far, known methods for detecting a biomolecule on a substrate comprise detecting a hybridisation event between a target biomolecule and a probe, for example by detecting the formation of a duplex between a nucleic acid target sequence and a nucleic acid probe or a probe and its target (WO 02/069653). Other detection methods have been described for example by Ford in “A biologically relevant optical sensor based on the hybridization of a fluorescent oligonucleotide probe with surface immobilized DNA” (Biotechnology and Bioengineering Vol. 95, No. 1, 2002).
Also known are methods for treatment of a substrate comprising immobilising at least a biomolecule on at least a part of the substrate, detecting free, i.e. unbound and unbound as a result of detection, biomolecule, and optionally delivering the biologically active biomolecule to the substrate.
It is known to treat substrates by attaching a nucleic acid on its surface.
US 2007/0204242 describes a nucleic acid hybridisation assay method for detecting and quantifying a nucleic acid of interest on or in a sample. The method is characterised in that a nucleic acid on the surface of a biosensor is contacted with a test sample comprising a nucleic acid of interest. The nucleic acid of interest may be attached to a naturally occurring nucleic acid, such as a DNA or a RNA, as a result of, for example, a hybridisation event. In a next step, the presence of the nucleic acid of interest is detected by detecting a subsequent hybridisation event.
U.S. Pat. No. 5,744,311 describes a method for detection and/or quantification of analytes, especially nucle
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