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Title:
METHOD FOR TESTING FOR TISSUE-SPECIFIC CELL DAMAGE
Document Type and Number:
WIPO Patent Application WO/2019/207921
Kind Code:
A1
Abstract:
Provided is a method for testing for a tissue-specific cell damage, the method being capable of detecting a target tissue-specific cell damage by amplification, by an at least two-stage ARMS-PCR, of a target area that includes not more than 150 bases and has at least three characteristic or specific nonmethylated cytosines in DNA of a target tissue or cells in the blood of a subject. Prior to the ARMS-PCR amplification step, the method may include a step of changing the nonmethylated cytosine of the cfDNA to a base other than cytosine, and may further include a step of ensuring the PCR amplification, regardless of the presence or absence of methylation, of a region that contains the target area of the cfDNA. The assessment that the target tissue or cells have been injured can be made based on confirmation that the target area has been amplified by ARMS-PCR, and the degree of injury to the target tissue or cells can be assessed by quantitation of the amount of target region-bearing DNA in the amplification product.

Inventors:
MATSUHISA MUNEHIDE (JP)
KURODA AKIO (JP)
YAMADA MISUZU (JP)
Application Number:
JP2019/005850
Publication Date:
October 31, 2019
Filing Date:
February 18, 2019
Export Citation:
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Assignee:
MATSUHISA MUNEHIDE (JP)
International Classes:
C12Q1/686; C12Q1/6883; C12N15/09
Domestic Patent References:
WO2011132798A12011-10-27
WO2009104714A12009-08-27
WO2013089063A12013-06-20
Foreign References:
CN106947805A2017-07-14
JP2017511142A2017-04-20
Other References:
NEIMAN,D ET AL.: "Islet cells share promoter hypomethylation independently of expression, but exhibit cell -type-specific methylation in enhancers", PNAS, vol. 114, no. 51, 2017, pages 13525 - 13530, XP055647838
OLSEN, J. A. ET AL.: "Circulating Differentially Methylated Amylin DNA as a Biomarker of beta - Cell Loss in Type 1 Diabetes", PLOS ONE, vol. 11, no. 4, 2016, pages 1 - 15, XP055346200, DOI: 10.1371/journal.pone.0152662
KATO,KIKUYA: "Technologies for detecting circulating tumor DNA", CYTOMETRY RESEARCH, vol. 26, no. 1, 2016, pages 7 - 12, XP055647835
KURODA,A ET AL.: "Detection of Pancreatic Beta- Cell DNA in the Circulation Using the Dual Amplification Refractory Mutation System PCR", AMERICAN DIABETES ASSOCIATION, vol. 67, no. Suppl. 1, July 2018 (2018-07-01), pages 2102-P
Attorney, Agent or Firm:
TADA Eiko et al. (JP)
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