The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Date of Patent:
Nov. 29, 2016

Filed:

Nov. 25, 2015
Applicant:

On-chip Biotechnologies Co., Ltd., Tokyo, JP;

Inventors:

Kazuo Takeda, Tokyo, JP;

Fumie Jimma, Tokyo, JP;

Masashi Takao, Miyagi, JP;

Attorney:
Primary Examiner:
Int. Cl.
CPC ...
G01N 15/14 (2006.01); G01N 21/64 (2006.01); G01N 33/53 (2006.01); G01N 33/574 (2006.01); G01N 33/543 (2006.01); G01N 1/30 (2006.01); G01N 15/10 (2006.01);
U.S. Cl.
CPC ...
G01N 33/57492 (2013.01); G01N 1/30 (2013.01); G01N 15/1436 (2013.01); G01N 15/1459 (2013.01); G01N 21/6428 (2013.01); G01N 33/54333 (2013.01); G01N 33/574 (2013.01); G01N 2015/1006 (2013.01); G01N 2015/1477 (2013.01); G01N 2021/6439 (2013.01); G01N 2201/06113 (2013.01); G01N 2333/705 (2013.01); G01N 2333/70596 (2013.01);
Abstract

Conventional CTC detection methods have been problematic in that 1) there is no technique for automatically determining and counting live CTCs in a brief period of time, 2) no process has been developed for detecting, counting, and thereafter collecting and culturing live CTCs, and 3) there exists no flow cytometer that is contamination free and is capable of measuring an entire sample. Provided is a CTC detection method which comprises a pre-treatment step for concentrating and fluorescence staining CTCs, and a step for identifying and counting CTCs. The pre-treatment step includes attaching magnetic beads to EpCAM antibodies expressed by epithelial cell-derived CTCs and concentrating the CTCs through the use of a magnet, fluorescently labeling an epithelia cell surface marker of the CTCs through the use of EpCAM antibodies or 5E11 antibodies, and performing two types of nuclear staining, one being cell membrane-permeable and the other being cell membrane-impermeable. The identifying and counting step includes evaluating the respective absolute concentrations of live and dead CTCs in a volume of blood by automatically identifying CTCs by the ratio of a plurality of fluorescence signal intensities using a flow cytometer, and differentiating between and counting the live CTCs and the dead CTCs. In the cytometer, an entire liquid-feeding system that includes a flow cell can be replaced for each sample, and the total amount of a liquid sample can be measured.


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