Assays

Abstract

A device comprising a rigid substrate, a flexible cover element at least partially covering the substrate, a first structure formed in the substrate, adapted for accommodating liquids and adapted for releasing contents of one or more cells, spores, or viruses, the contents including the target molecules, a second structure formed in the substrate, adapted for accommodating liquids and comprising at least one binding member adapted for capturing the target molecules and for determining a value indicative of the presence and/or amount of the target molecules, a micro fluidic network interconnecting at least the first structure and the second structure, and an actuator member adapted for effecting a fluid flow between the first structure and the second structure by pressing the flexible cover element against the substrate to selectively close a portion of the micro fluidic network.

1 - 256 . (canceled) 
     
     
         257 . A method for detecting a human immunodeficiency virus (HIV) target nucleic acid in a liquid, comprising:
 a) introducing a liquid into a reaction chamber of a microfluidic device, the liquid comprising an HIV polynucleotide, binding the HIV polynucleotide to a first binding member located within the reaction chamber;   b) amplifying the HIV polynucleotide within the reaction chamber to form amplicons, the amplification being performed in the presence of a reporter compound, binding a subset of the reporter compound to amplicons within the reaction chamber, binding a remaining subset of the reporter compound to a second binding member within the reaction chamber, and detecting the remaining subset of the reporter compound bound to the second binding member; and   c) determining a value indicative of the presence and/or amount of reporter compound captured on the second binding member; and determining a value indicative of the amount of target nucleic acid based on the value indicative of the amount of reporter compound captured on the second binding member.   
     
     
         258 . The method of  claim 257 , wherein said HIV polynucleotide is HIV RNA. 
     
     
         259 . The method of  claim 258 , wherein said amplifying is RT-PCR. 
     
     
         260 . The method of  claim 257 , wherein said HIV polynucleotide is HIV DNA. 
     
     
         261 . The method of  claim 257 , wherein said HIV polynucleotide is free or cell-associated. 
     
     
         262 . The method of  claim 257 , wherein said first binding member and said second binding members are solid supports. 
     
     
         263 . The method of  claim 262 , wherein said first binding members is a latex bead or a magnetic bead. 
     
     
         264 . The method of  claim 262 , wherein said second binding member is an array surface. 
     
     
         265 . The method of  claim 264 , wherein the array surface comprises one or more different reporter specific capture molecules being capable of capturing a reporter compound on the second binding member. 
     
     
         266 . The method of  claim 257 , wherein said reporter compound is an oligonucleotide. 
     
     
         267 . The method of  claim 266 , wherein said oligonucleotide is labeled.