Nmr systems and methods for the rapid detection of analytes

Abstract

This invention features systems and methods for the detection of analytes, and their use in the treatment and diagnosis of disease.

1 . An amplified lysate solution produced by a method for amplifying a target nucleic acid in a whole blood sample, the method comprising:
 (a) providing a first extract produced by lysing the red blood cells in a whole blood sample from a subject, centrifuging the sample to form a supernatant and a pellet, discarding some or all of the supernatant, and resuspending the pellet to form a second extract;   (b) lysing cells in the second extract to form a lysate; and   (c) providing the lysate of step (b) in a detection tube and amplifying nucleic acids therein to form an amplified lysate solution comprising from 40% (w/w) to 95% (w/w) the target nucleic acid and from 5% (w/w) to 60% (w/w) nontarget nucleic acid.   
     
     
         2 . The amplified lysate solution of  claim 1 , wherein said amplified lysate solution comprises whole blood proteins and non-target oligonucleotides. 
     
     
         3 . A kit for amplifying a target nucleic acid, said kit comprising:
 a detection tube   a forward primer and a reverse primer, each of which is universal to multiple  Candida  species to form a solution comprising a  Candida  amplicon, and   instructions for:
 (a) providing a first extract produced by lysing the red blood cells in a whole blood sample from a subject, centrifuging the sample to form a supernatant and a pellet, discarding some or all of the supernatant, and resuspending the pellet to form a second extract; 
 (b) lysing cells in the second extract to form a lysate; and 
 (c) providing the lysate of step (b) in a detection tube and amplifying nucleic acids therein to form an amplified lysate solution comprising from 40% (w/w) to 95% (w/w) the target nucleic acid and from 5% (w/w) to 60% (w/w) nontarget nucleic acid. 
   
     
     
         4 . A composition, comprising:
 (a) portion of an extract from a whole blood sample suspected of containing a pathogen prepared by i) lysing the red blood cells, ii) discarding some or all of the supernatant, and iv) lysing any residual cells to form the extract;   (b) a forward primer;   (c) a reverse primer;   (d) a thermal stable polymerase; and   (e) deoxynucleotide triphosphates, buffer, and magnesium.   
     
     
         5 . The composition of  claim 4 , wherein said pellet is washed prior to said (iv) lysing of any residual cells to form an extract. 
     
     
         6 . The composition of  claim 4 , wherein said (ii) centrifuging the sample and said (iii) discarding of some of all of the supernatant is repeated prior to said (iv) lysing any residual cells to form an extract.