	US 12,007,390 B2
	Prefabricated microparticle for performing a detection of an analyte
Katrin Steinmetzer, Jena (DE); Stephan Hubold, Jena (DE); Thomas Ellinger, Jena (DE); Eugen Ermantraut, Jena (DE); and Torsten Schulz, Jena (DE)
Assigned to BLINK AG, Jena (DE)
Filed by BLINK AG, Jena (DE)
Filed on Jun. 21, 2021, as Appl. No. 17/352,958.
Application 17/352,958 is a continuation of application No. 16/475,109, granted, now 11,073,518, previously published as PCT/EP2017/084370, filed on Dec. 22, 2017.
Claims priority of application No. 16207455 (EP), filed on Dec. 30, 2016.
Prior Publication US 2021/0318303 A1, Oct. 14, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 33/543 (2006.01); C12Q 1/6848 (2018.01)

CPC G01N 33/54393 (2013.01) [C12Q 1/6848 (2013.01); G01N 33/54313 (2013.01)]

20 Claims

1. A method of performing a detection of an analyte in a sample, said method comprising the steps:

a) providing prefabricated microparticles, each of which has a surface and includes a void volume for receiving an aqueous solution, wherein each of said prefabricated microparticles is dispersible in a non-aqueous medium and, upon dispersion in a non-aqueous medium, provides for a defined reaction space in such non-aqueous medium, in which defined reaction space a chemical or biochemical reaction indicating the presence of an analyte can be performed, and wherein each of said prefabricated microparticles does not comprise a capture agent that selectively and specifically binds the analyte to be detected; wherein each of said microparticles is a porous microparticle and is capable of taking up liquid and an analyte present in said liquid, from surroundings to which it is exposed; and wherein each of said prefabricated microparticles further comprises a detection agent that is specific for the analyte, and that binds said analyte;

b) exposing said prefabricated microparticles to an aqueous sample suspected of containing an analyte to be detected, thus allowing the uptake of liquid and of an analyte present in said liquid, in the porous microparticles, i.e. in the space provided for by the pores of said microparticles; and

c) placing the prefabricated microparticles into a non-aqueous phase, and using the void volume of each of said prefabricated microparticles as a defined reaction space in which a chemical or biochemical reaction indicating the presence of an analyte, is performed, by either

d1) detecting the detection agent bound to said analyte;

d2) amplifying the analyte, if present, by way of an amplification reaction, and detecting the thus amplified product by means of said detection agent, wherein said analyte is a nucleic acid and said amplification reaction is a nucleic acid amplification,

d3) performing a signal amplification reaction and detecting the thus amplified signal,

wherein said reaction space in which said chemical or biochemical reaction indicating the presence of an analyte, is performed, is defined by said void volume of each of said prefabricated microparticles and is not larger than said void volume of each of said prefabricated microparticles.