Assays and Devices For Identifying Pathogens
Provided are methods and devices for identifying pathogens based on protein translation from ribosomes isolated from the pathogens.
1 . A method, comprising:
(a) contacting a support comprising a binding agent capable of binding a ribosome with a sample comprising ribosomes;
(b) washing the support after contact with the sample;
(c) incubating the support in the presence of a solution comprising mRNA and amino acids under conditions suitable to allow ribosomes bound to the binding agent to translate the mRNA in the solution into a polypeptide; and
(d) determining the presence of the polypeptide.
2 . The method of claim 1 , wherein the ribosomes are specific to a pathogen and the method further comprises determining the presence of the pathogen in the sample based on the presence of the polypeptide.
3 . The method of claim 2 , wherein the pathogen is a bacterium.
4 . The method of claim 1 , wherein the binding agent is capable of specifically binding a ribosome.
5 . The method of claim 3 , wherein the ribosomes are bacterial ribosomes and the method further comprises releasing the ribosomes from at least one bacterium.
6 . The method of claim 1 , wherein the support is a porous material.
7 . The method of claim 3 , wherein the porous material is nitrocellulose.
8 . The method of claim 1 , wherein the binding agent is an antibody.
9 . The method of claim 1 , wherein the binding agent is an agent capable of binding ribosomal RNA.
10 . The method of claim 9 , wherein the agent capable of binding ribosomal RNA is a nucleic acid.
11 . The method of claim 1 , wherein the solution comprising mRNA and amino acids further comprises a substrate.
12 . The method of claim 11 , wherein the polypeptide is an enzyme.
13 . The method of claim 12 , wherein determining the polypeptide comprises determining the presence of a reaction product resulting from reaction of the enzyme with a substrate for the enzyme.
14 . The method of claim 13 , wherein determining the polypeptide comprises optically detecting the presence of the reaction product.
15 . A method, comprising:
(a) capturing ribosomes on a support using a binding agent capable of binding the ribosomes;
(b) incubating the captured ribosomes with a solution comprising mRNA and amino acids; and
(c) detecting the polypeptide produced by the incubated, captured ribosomes.
16 . A device, comprising a support comprising a binding agent capable of binding a ribosome.
17 . The device of claim 16 , wherein the support is a porous material.
18 . The device of claim 17 , wherein the porous material is nitrocellulose.
19 . The device of claim 16 , wherein the binding agent is an antibody.
20 . The device of claim 16 , wherein the binding agent is an agent capable of binding ribosomal RNA.
21 . The device of claim 20 , wherein the agent capable of binding ribosomal RNA is a nucleic acid.