Method of Preparing Nucleic Acids for Detection

Abstract

A method is provided for preparing a test sample for detecting a predetermined target nucleic acid. The method includes the steps of providing a test probe comprising an oligonucleotide attached to a nanoparticle and providing a hybridization unit containing the test sample and the test probe, wherein said hybridization unit further includes a target sample substrate and a distribution manifold coupled to a first side of the substrate. The method further includes the steps of clamping a processing fluids manifold to the distribution manifold of the hybridization unit, denaturing the test sample and preparing the test sample for detecting the predetermined target nucleic acid by pumping a plurality of processing fluids between the processing fluids source manifold and distribution manifold to hybridize the test probe and predetermined target nucleic acid to the target sample substrate, to wash the hybridized sample and to amplify a detectable parameter of the hybridized sample.

1 . A method of preparing a test sample for purposes of detecting a predetermined target nucleic acid, such method comprising the steps of:
 (a) providing a test probe comprising an oligonucleotide attached to a nanoparticle;   (b) providing a hybridization unit containing the test sample and the test probe, wherein said hybridization unit further comprises a target sample substrate and a distribution manifold coupled to a first side of the substrate;   (c) clamping a processing fluids manifold to the distribution manifold of the hybridization unit;   (d) denaturing the test sample; and   (e) preparing the test sample for detecting the predetermined target nucleic acid by pumping a plurality of processing fluids between the processing fluids source manifold and distribution manifold to hybridize the test probe and predetermined target nucleic acid to the target sample substrate, to wash the hybridized sample and to amplify a detectable parameter of the hybridized sample.   
     
     
         2 . The method of preparing the test sample as in  claim 1  further comprising disposing a hybridization solution into a sample well of the hybridization unit. 
     
     
         3 . The method of preparing the test sample as in  claim 2  further comprising disposing the test probe into the sample well. 
     
     
         4 . The method of preparing the test sample as in  claim 1  further comprising disposing the test sample in the sample well. 
     
     
         5 . The method of preparing the test sample as in  claim 4  wherein the step of denaturing the test sample further comprises heating the sample well. 
     
     
         6 . The method of preparing the test sample as in  claim 5  further comprising disposing an oligonucleotide having a sequence complementary to a first portion of a genetic sequence of the predetermined target nucleic acid within a hybridization zone of the hybridization unit. 
     
     
         7 . The method of preparing the test sample as in  claim 6  wherein the step of disposing the oligonucleotide within a hybridization zone of the hybridization unit further comprises connecting the oligonucleotide to the target sample substrate. 
     
     
         8 . The method of preparing the test sample as in  claim 7  further comprising defining the oligonucleotide attached to the nanoparticle as having a genetic sequence complementary to a second portion of the genetic sequence of the predetermined target nucleic acid. 
     
     
         9 . The method of preparing the test sample as in  claim 8  further comprising drawing a content of the sample well from the sample well into the hybridization zone using a fluid coupled through a first port of the processing fluids manifold. 
     
     
         10 . The method of preparing the test sample as in  claim 9  further comprising chilling the content of the sample well as the content is drawn into the hybridization zone. 
     
     
         11 . The method of preparing the test sample as in  claim 9  further comprising hybridizing the probe and predetermined nucleic acid with the oligonucleotide connected to the target sample substrate by shuttling the content of the well through the hybridization zone a predetermined number of times. 
     
     
         12 . The method of preparing the test sample as in  claim 11  further comprising flushing the hybridized probe, predetermined nucleic acid and first oligonucleotide by introducing wash fluid through a second port and discharging wash fluid through the first port. 
     
     
         13 . The method of preparing the test sample as in  claim 12  further comprising amplifying optical characteristics of the hybridized probe, predetermined nucleic acid and first oligonucleotide by introducing a plating solution through a second port and discharging spent plating solution through the first port. 
     
     
         14 . The method of preparing the test sample as in  claim 13  further comprising defining the plating solution as being a silver solution. 
     
     
         15 . A method of preparing a plurality of test samples for purposes of detecting predetermined target nucleic acids, such method comprising the steps of:
 (a) providing a hybridization unit containing the plurality of test samples, said hybridization unit further comprising a target samples substrate and a distribution manifold coupled to a first side of the substrate said target samples;   (b) clamping a processing fluids manifold to the distribution manifold of the hybridization unit;   (c) denaturing the test samples; and   (d) preparing the plurality of test samples for detecting the predetermined nucleic acids by pumping a plurality of processing fluids between the processing fluids source manifold and distribution manifold to hybridize the predetermined target nucleic acid to the target samples substrate, to wash the hybridized samples and to amplify a detectable parameter of the hybridized samples.   
     
     
         16 . The method of preparing the test sample as in  claim 15  further comprising disposing the plurality of test samples in a plurality of respective sample wells. 
     
     
         17 . The method of preparing the test sample as in  claim 16  wherein the step of denaturing the test samples further comprises heating a sample well of the plurality of respective sample wells. 
     
     
         18 . The method of preparing the test sample as in  claim 17  further comprising disposing a liquid test probe into a first well of the plurality of sample wells. 
     
     
         19 . The method of preparing the test sample as in  claim 18  further comprising disposing an first oligonucleotide having a sequence complementary to a first portion of a sequence of a nucleic acid of the predetermined nucleic acids within a hybridization zone of the hybridization unit. 
     
     
         20 . The method of preparing the test sample as in  claim 19  wherein the step of disposing the first oligonucleotide within a hybridization zone of the hybridization unit further comprises connecting an end of the oligonucleotide to the target samples substrate. 
     
     
         21 . The method of preparing the test sample as in  claim 20  wherein the test probe further comprises a nanoparticle and a second oligonucleotide disposed on a surface of the nanoparticle, said second oligonucleotide having a sequence complementary to a second portion of the sequence of the nucleic acid of the predetermined nucleic acids. 
     
     
         22 . The method of preparing the test sample as in  claim 21  further comprising drawing a content of a well of the plurality of wells from the well into the hybridization zone using a fluid coupled through a first port of the processing fluids manifold. 
     
     
         23 . The method of preparing the test sample as in  claim 22  further comprising cooling the content of the sample well of the sample wells as the content is drawn into the hybridization zone. 
     
     
         24 . The method of preparing the test sample as in  claim 22  further comprising hybridizing the probe and predetermined nucleic acid with the first oligonucleotide by shuttling the content of the well through the hybridization zone a predetermined number of times to mix the probe and predetermined nucleic acid with the first oligonucleotide. 
     
     
         25 . The method of preparing the test sample as in  claim 24  further comprising flushing the hybridized probe, predetermined nucleic acid and first oligonucleotide by introducing wash fluid through a second port and discharging wash fluid through the first port. 
     
     
         26 . The method of preparing the test sample as in  claim 25  further comprising amplifying optical characteristics of the hybridized probe, predetermined nucleic acid and first oligonucleotide by introducing a plating solution through a second port and discharging spent plating solution through the first port. 
     
     
         27 . The method of preparing the test sample as in  claim 26  further comprising defining the plating solution as being a silver solution.