Rapid genotyping analysis and the device thereof

Abstract

The present invention describes methods of performing rapid nucleic acid detection using a flow through process. The methods comprise single-step signal amplification and/or a one-step hybridization protocol. Using the flow through hybridization process, the present invention provides a more efficient, faster and less expensive genotyping method. This invention further provides a Single Nucleotide Polymorphism (SNP)-based DNA fingerprinting method for rapid and accurate genotyping, identification as well as DNA analyses of genetic materials from human beings and other different organisms. In addition this invention also discloses devices for rapid and sensitive analysis of target analysts.

1 . A method of performing rapid nucleic acid detection, comprising the steps of:
 (a) obtaining a sample comprising a target nucleic acid molecule;   (b) mixing the target nucleic acid molecule with (i) a first probe that will bind to the nucleic acid molecule, and (ii) a second agent that will bind to the first probe, thereby forming a nucleic acid molecule complex in solution;   (c) applying in a flow through manner the solution comprising the nucleic acid molecule complex to an array comprising a third probe that will bind to the complex, thereby capturing the nucleic acid molecule complex on the array; and   (d) detecting the captured nucleic acid molecule on the array.   
     
     
         2 . The method of  claim 1 , wherein the target nucleic acid molecule is of human, bacterial, or viral origin. 
     
     
         3 . The method of  claim 1 , wherein the human is having or is suspected to have cancer. 
     
     
         4 . The method of  claim 1 , wherein the target nucleic acid molecule is obtained without amplification. 
     
     
         5 . The method of  claim 1 , wherein the first probe and the third probe bind to different regions of the target nucleic acid molecule. 
     
     
         6 . The method of  claim 1 , wherein the third probe is an allelic-specific oligonucleotide probe. 
     
     
         7 . The method of  claim 6 , wherein the allelic-specific oligonucleotide probe is generated from nucleic acid sequence data bank. 
     
     
         8 . The method of  claim 1 , wherein the third probe can identify single nucleotide polymorphism of the target nucleic acid molecule. 
     
     
         9 . The method of  claim 1 , wherein the detection of captured nucleic acid molecule is performed by fluorescence tags, quantum dot labeling, colloidal gold particle labeling, magnetic particle labeling, or enzyme-linked substrate assay. 
     
     
         10 . A method of performing rapid nucleic acid detection, comprising the steps of:
 (a) obtaining a sample comprising a target nucleic acid molecule;   (b) mixing the target nucleic acid molecule with (i) a first probe that will bind to the nucleic acid molecule, (ii) a first antibody, and (iii) a labeling agent, wherein the first probe will form a complex with the target nucleic acid molecule, and the first antibody will bind to the resulting complex;   (c) applying in a flow through manner a solution comprising the nucleic acid molecule complex to an array comprising a second antibody that will bind to the first antibody, thereby capturing the nucleic acid molecule complex on the array; and   (d) detecting the captured nucleic acid molecule on the array.   
     
     
         11 . The method of  claim 10 , wherein the target nucleic acid molecule is of human, bacterial, or viral origin. 
     
     
         12 . The method of  claim 11 , wherein the human is having or is suspected to have cancer. 
     
     
         13 . The method of  claim 10 , wherein the target nucleic acid molecule is obtained without amplification. 
     
     
         14 . The method of  claim 10 , wherein the first probe and the labeling agent bind to different regions of the target nucleic acid molecule. 
     
     
         15 . The method of  claim 10 , wherein the detection of captured nucleic acid molecule is performed by fluorescence tags, quantum dot labeling, colloidal gold particle labeling, magnetic particle labeling, or enzyme-linked substrate assay. 
     
     
         16 . A device capable of practicing the method of  claim 1 . 
     
     
         17 . A lateral flow hybridization device comprising:
 (a) a membrane for immobilizing capture molecules capable of capturing target analysts, wherein the membrane is positioned in an reaction chamber where it can be maintained in controlled conditions;   (b) controlling elements that can be regulated to maintain the reaction chamber in the controlled conditions;   (c) connecting elements for connection to a power supply and control unit that can regulate and maintain the controlled conditions; and   (d) liquid delivery elements capable of accepting and removing solution to and from the reaction chamber, wherein the solution is maintained in a lateral flow direction that allows the solution flows through the membrane pores in cross-section so that all target analysts pass through the capture molecules immobilized on the membrane, thereby providing higher sensitivity of analysts detection.   
     
     
         18 . The device of  claim 17 , wherein the power supply and control unit is capable of supplying energy and providing regulatory control to maintain the reaction chamber in the controlled conditions. 
     
     
         19 . The device of  claim 17 , wherein the controlled conditions comprise accurate control of temperature within 0.5 degree C. 
     
     
         20 . The device of  claim 17 , wherein the controlling elements are heating and cooling elements. 
     
     
         21 . The device of  claim 20 , wherein the heating and cooling elements are Peltier heating and cooling elements or elements for circulating liquid or air. 
     
     
         22 . The device of  claim 20 , wherein the heating and cooling elements comprise heat conduction plate, heating and cooling sensor, heat sink, and heating and cooling fan. 
     
     
         23 . The device of  claim 17 , wherein the membrane is formed of material containing nitrocellulose, nylon, Nytron, Biodyne, or Porex. 
     
     
         24 . The device of  claim 17 , wherein accepting and removing solution to and from the reaction chamber is by regulated liquid pumping. 
     
     
         25 . The device of  claim 24 , wherein the liquid pumping is used to recirculate solution containing the target analysts through the membrane. 
     
     
         26 . A lateral flow hybridization system comprising more than one of the hybridization device of  claim 17 , wherein the hybridization devices are connected to a power supply and control unit capable of supplying energy and providing regulatory control to the hybridization devices. 
     
     
         27 . The system of  claim 26 , wherein each of the hybridization devices is controlled independently by the power supply and control unit so that each hybridization device can perform different analysis under different conditions. 
     
     
         28 . A lateral flow hybridization device comprising:
 (a) more than one reaction chambers, each of which comprises a membrane for immobilizing capture molecules capable of capturing target analysts;   (b) controlling elements that can be regulated to maintain the reaction chamber in controlled conditions;   (c) connecting elements for connection to a power supply and control unit that can regulate and maintain the controlled conditions; and   (d) liquid delivery elements capable of accepting and removing solution to and from the reaction chambers, wherein the solution is maintained in a lateral flow direction that allows the solution flows through the membrane pores in cross-section so that all target analysts pass through the capture molecules immobilized on the membrane, thereby providing higher sensitivity of analysts detection.   
     
     
         29 . The device of  claim 28 , wherein the power supply and control unit is capable of supplying energy and providing regulatory control to maintain the reaction chambers in the controlled conditions. 
     
     
         30 . The device of  claim 28 , wherein the controlled conditions comprise accurate control of temperature within 0.5 degree C. 
     
     
         31 . The device of  claim 28 , wherein the controlling elements are heating and cooling elements. 
     
     
         32 . The device of  claim 31 , wherein the heating and cooling elements are Peltier heating and cooling elements or elements for circulating liquid or air. 
     
     
         33 . The device of  claim 28 , wherein the membrane is formed of material containing nitrocellulose, nylon, Nytron, Biodyne, or Porex. 
     
     
         34 . The device of  claim 28 , wherein accepting and removing solution to and from the reaction chamber is by regulated liquid pumping. 
     
     
         35 . The device of  claim 28 , wherein the reaction chambers are disposable. 
     
     
         36 . The device of  claim 28 , wherein each of the reaction chambers is a separate unit.