Sysmex Corporation

The University of Tokyo

Ayako Sakai

Masahiro Kajita

Hideki Ishihara

Noriko Oka

Noriaki Yamamoto

Hiroyuki Aburatani

Kaya Tai

Yutaka Midorikawa

Genta Nagae

Yoshihiko Tashima

Atsushi Kaneda

Kouichi Yagi

The present invention relates to a hybridoma producing an anti-methylated DNA antibody, obtained by cell fusion of an antibody-producing cell obtained from an animal immunized with an antigen containing 5'-(5-methyl-2&#x2032;-deoxycytidine-3&#x2032;-phospho)-2&#x2032;-deoxyguanosine 3&#x2032;-phosphate with a myeloma cell. The present invention also relates to a monoclonal antibody produced by the hybridoma and a method for immunoprecipitation of a methylated DNA using the antibody.

Hybridoma producing anti-methylated DNA antibody and utilization of same

The present invention relates to a method for determination of the presence or absence of cancer cells in a biological sample or a method for determination of the prognosis of a colorectal cancer patient based on a result obtained by extracting DNA from a biological sample and analyzing methylation status of a marker gene in the DNA.

Methylation analysis to determine the presence of cancer cells

Success or failure of unmethylated cytosine conversion treatment is determined by using a first primer set comprising plural primers that hybridize with a nucleic acid comprising a nucleotide sequence not containing cytosine in the nucleotide sequence of biological DNA that is subject to an unmethylated cytosine conversion treatment and a second primer set comprising plural primers that hybridize with a nucleic acid comprising a nucleotide sequence in which cytosine in a nucleotide sequence containing cytosine and not containing a CpG site is converted into a base other than cytosine, in the nucleotide sequence of biological DNA.

Kit and method for determining whether or not unmethylated cytosine conversion treatment is properly carried out and method for analyzing methylated DNA using the same

The method for detecting a methylated cytosine of the present invention comprises the steps of:

Method of detecting methylated cytosine

The present invention provides a method of analyzing methylated DNA, comprising steps of: (A) treating a DNA-containing sample with a restriction enzyme to obtain a sample containing a DNA fragment; (B) concentrating methylated DNA contained in the sample obtained in step (A) to obtain a methylated DNA concentrate; (C) subjecting the methylated DNA concentrate obtained in step (B) and a primer set to nucleic acid amplification reaction, wherein the primer set performs the nucleic acid amplification reaction in step (C) by using a template DNA which does not have a CpG site; (D) detecting an amplification product obtained in step (C); (E) judging whether the methylated DNA concentrate obtained in step (B) is appropriate as a sample for detection of methylated DNA, on the basis of the detection result of the amplification product in step (D); and (F) analyzing the methylated DNA contained in the methylated DNA concentrate.

Growing community of inventors

Kobe, Japan

Ayako Sakai