Methods and devices for detection and acquisition of biomarkers

Abstract

The present invention provides devices and methods for detecting and capturing molecular biomarkers from a subject in situ. Specifically, the devices contain an array of microneedles to which are attached probes specific for one or more biomarkers of interest. The devices can be used directly on a subject (e.g., via skin piercing) in detecting the biomarkers in the body of the subject (e.g., tissues, blood stream).

1 . A device for detecting or extracting one or more nucleic acid biomarkers from an in situ tissue or biological sample of a subject, wherein the device comprises two or more microneedles and one or more polynucleotide probes specific for the nucleic acid biomarkers, wherein the polynucleotide probes are attached to the two or more microneedles and a center-to-center distance between the two or more microneedles is greater than 200 μm and less than 1000 μm. 
     
     
         2 . The device of  claim 1 , comprising a plurality of polynucleotide probes specific for a plurality of different biomarkers, wherein each polynucleotide probe is attached to a different microneedle. 
     
     
         3 . The device of  claim 1 , comprising one polynucleotide probe for a specific biomarker, wherein the polynucleotide probe is attached to the two or more microneedles. 
     
     
         4 . The device of  claim 1 , wherein the microneedles are made of a polymer, a metal or a ceramic. 
     
     
         5 . The device of  claim 1 , wherein the polynucleotide probes are polynucleotides complementary to the nucleic acid biomarkers. 
     
     
         6 . The device of  claim 5 , wherein the microneedles are polycarbonate, and the polynucleotide probes are attached to the microneedles via a thiol/amino bifunctional linker. 
     
     
         7 . (canceled) 
     
     
         8 . The device of  claim 1 , wherein the polynucleotide probes are attached to the microneedles via a linker as a spacer or to stabilize orientation of the polynucleotide probes. 
     
     
         9 . The device of  claim 1 , wherein the two or more microneedles are formed on a substrate. 
     
     
         10 . The device of  claim 1 , further comprising reagents for amplifying and identifying the nucleic acid biomarkers captured by the polynucleotide probes. 
     
     
         11 . A method for detecting one or more biomarkers from an in situ tissue or biological sample in a subject, comprising (a) providing the device of  claim 1 ; (b) contacting the device of  claim 1  with the in situ tissue or biological sample of the subject such that the nucleic acid biomarkers are bound to the polynucleotide probes; and (c) detecting the nucleic acid biomarkers that are bound to the polynucleotide probes. 
     
     
         12 . The method of  claim 11 , wherein the device comprises a plurality of polynucleotide probes specific for a plurality of different biomarkers, and wherein each polynucleotide probe is attached to a different microneedle. 
     
     
         13 . The method of  claim 11 , wherein the device comprises one polynucleotide probe specific for a biomarker, and wherein the polynucleotide probe is attached to the two or more microneedles. 
     
     
         14 . The method of  claim 11 , wherein the microneedles are made of a polymer, a metal or a ceramic. 
     
     
         15 . The method of  claim 11 , wherein the polynucleotide probes are polynucleotides complementary to the nucleic acid biomarkers. 
     
     
         16 . The method of  claim 15 , wherein the nucleic acid biomarkers bound to the polynucleotide probes on the microneedles are detected by PCR. 
     
     
         17 . The method of  claim 15 , wherein the microneedles are polycarbonate, and the polynucleotide probes are attached to the microneedles via a linker. 
     
     
         18 . (canceled) 
     
     
         19 . (canceled) 
     
     
         20 . The method of  claim 11 , wherein the microneedles are formed on a substrate. 
     
     
         21 . The method of  claim 11 , wherein the tissue is the subject's blood stream, and the microneedle device is contacted with the blood stream by piercing the skin of the subject. 
     
     
         22 . (canceled) 
     
     
         23 . The method of  claim 1 , wherein the two or more microneedles are solid. 
     
     
         24 . The method of  claim 1 , wherein the polynucleotide probes comprise DNA.