	US 11,834,700 B2
	Method for analyzing target particle, analytical reagent, and analyzer
Yoshiaki Sugizaki, Fujisawa Kanagawa (JP)
Assigned to Kabushiki Kaisha Toshiba, Tokyo (JP)
Filed by KABUSHIKI KAISHA TOSHIBA, Tokyo (JP)
Filed on Sep. 13, 2021, as Appl. No. 17/447,469.
Claims priority of application No. 2021-045863 (JP), filed on Mar. 19, 2021.
Prior Publication US 2022/0298544 A1, Sep. 22, 2022
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/6806 (2018.01); G01N 33/543 (2006.01); C12N 9/22 (2006.01)

CPC C12Q 1/6806 (2013.01) [C12N 9/22 (2013.01); G01N 33/54326 (2013.01)]

18 Claims

1. A method for analyzing a target particle in a specimen, the method comprising:

a) dispensing the specimen into first₁to first_ncontainers configured to capture the target particle;

b) removing a contaminant from the specimen, the contaminant being other than the target particle to be captured;

c) cladding first to m-th probes to each container of the first₁to first_ncontainers, each of the first to m-th probes having at least a binding portion and a nucleic acid reporter portion,

the binding portion of each of the first to m-th probes respectively binding to first to m-th surface markers of the target particle;

the nucleic acid reporter portion of each of the first to m-th probes respectively including a common amplification sequence to which a common primer set binds and a specific amplification sequence to which a specific primer set binds disposed in a region amplified by the common amplification sequence;

d) removing excessive probes of the first to m-th probes that have not bound to the target particle;

e) individually amplifying the reporter portion for each of the first₁to first_ncontainers using the common primer set to obtain first to n-th amplification products;

f) removing an excessive common primer set from the first to n-th amplification products;

g) dispensing the first to n-th amplification products into respective second₁to second_mcontainers;

h) further amplifying the first to n-th amplification products in the second₁to second_mcontainers using the first to m-th specific primer sets, respectively; and

i) analyzing at least one of a presence, absence or type of the target particles captured in the first₁to first_ncontainers by determining the types of surface markers present in the first₁to first_ncontainers based on the presence or absence of the first to n-th amplification products by the first to m-th specific primer sets in the second₁to second_mcontainers,

wherein n and m are integers of 2 or more.