Rapid comparative genomic hybridization using acoustic surface waves
First Claim
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1. A method for performing comparative genomic hybridization to immobilized nucleic acid, comprising the following steps:
- a) performing a prehybridization step comprising contacting a solid support comprising one or more immobilized nucleic acid probe molecules under hybridization conditions with a non-specific blocking nucleic acid at a concentration of at least 0.9 mg/ml, said one or more immobilized nucleic acid probe molecules capable of hybridizing with a sequence complementary thereto;
b) hybridizing a test sample containing genomic nucleic acid target molecules comprising a first detectable fluorescent label and a reference sample containing genomic nucleic acid reference molecules comprising a second detectable fluorescent label to the solid support under hybridizing conditions;
c) applying acoustic surface waves during step a) and step b), wherein the acoustic surface waves are generated by an alternating-voltage electrical signal, such that the acoustic surface waves cause movement of the test sample and the reference sample through impulse transfer;
d) washing the solid support following step a), step b), or both step a) and b);
e) determining the relative hybridization of the test sample and the reference sample to one or more nucleic acid probes of the solid support by detecting the fluorescence from the detectable labels on the target nucleic acid molecules and the reference nucleic acid molecules, wherein the relative hybridization indicates the comparative hybridization of the test and reference samples.
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Abstract
Disclosed is a method for performing nucleic acid hybridization assays which involve the application of acoustic surface waves. The hybridization assays may be used for detecting and mapping chromosomal or genetic abnormalities associated with various diseases or associated with predisposition to various diseases. In a particular aspect, the present method relates to the use of rapid nucleic acid hybridization methods, such as comparative genomic hybridization (CGH), for comparing nucleic acid segments of one genome to corresponding nucleic acid segments in another genome(s).
72 Citations
12 Claims
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1. A method for performing comparative genomic hybridization to immobilized nucleic acid, comprising the following steps:
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a) performing a prehybridization step comprising contacting a solid support comprising one or more immobilized nucleic acid probe molecules under hybridization conditions with a non-specific blocking nucleic acid at a concentration of at least 0.9 mg/ml, said one or more immobilized nucleic acid probe molecules capable of hybridizing with a sequence complementary thereto; b) hybridizing a test sample containing genomic nucleic acid target molecules comprising a first detectable fluorescent label and a reference sample containing genomic nucleic acid reference molecules comprising a second detectable fluorescent label to the solid support under hybridizing conditions; c) applying acoustic surface waves during step a) and step b), wherein the acoustic surface waves are generated by an alternating-voltage electrical signal, such that the acoustic surface waves cause movement of the test sample and the reference sample through impulse transfer; d) washing the solid support following step a), step b), or both step a) and b); e) determining the relative hybridization of the test sample and the reference sample to one or more nucleic acid probes of the solid support by detecting the fluorescence from the detectable labels on the target nucleic acid molecules and the reference nucleic acid molecules, wherein the relative hybridization indicates the comparative hybridization of the test and reference samples. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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Current AssigneeQuest Diagnostics Investments Incorporated (Quest Diagnostics Incorporated)
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Original AssigneeQuest Diagnostics Investments Incorporated (Quest Diagnostics Incorporated)
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InventorsMcCaskill, Christopher, Mohammed, Mansoor S., Dzidic, Natasa
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Primary Examiner(s)Sisson; Bradley L
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Application NumberUS11/883,150Publication NumberTime in Patent Office2,063 DaysField of SearchNoneUS Class Current436/94CPC Class CodesC12Q 1/6832 Enhancement of hybridisatio...C12Q 1/6841 In situ hybridisationC12Q 2523/313 Irradiation, e.g. UV irradi...Y10S 436/80 Fluorescent dyes, e.g. rhod...Y10T 436/143333 Saccharide [e.g., DNA, etc.]
