公开(公告)号：EP3012326A4

公开(公告)日：2016-12-21

申请号：EP14775157

申请日：2014-03-26

申请人： NIPPON GENE CO LTD

发明人： KITANI MASAKAZU ,  MAKI FUMINORI ,  IZAWA MASAKI ,  KANAYAMA SHINJI ,  YONEDA YUKO

IPC分类号： C12Q1/68 ,  C12N15/09

CPC分类号： C12Q1/6827 ,  C12Q1/6806 ,  C12Q1/6818 ,  C12Q1/6853 ,  C12Q2525/155 ,  C12Q2525/301 ,  C12Q2531/119 ,  C12Q2565/101

摘要： By establishing a simple method for extracting a nucleic acid from a human specimen, and using the LAMP method, which is an isothermal gene amplification method showing superior quickness and convenience, with a concept of measurement different from the conventional ones, there is established a test method including steps up to detection with a measurement apparatus. There is provided a method for detecting a target site, which is an LAMP method using four kinds of primers, FIP, F3 primer, BIP, and B3 primer, which are designed on the basis of six regions of a template polynucleotide, F1 region, F2 region, F3 region, B1 region, B2 region, and B3 region, wherein: the template polynucleotide contains the target site, the four kinds of primers are designed so that the target site exists between the F1 region and the F2 region, or between the B1 region and the B2 region; a loop primer designed on the basis of a region between the F 1 region and the F2 region or between the B 1 region and the B2 region on the side on which the target site exists, and an oligonucleotide probe that can associate with a region including the target site are used, the loop primer and the probe are designed so that the 5' end of the loop primer and the 3' end of the probe associate with the template polynucleotide at positions close to each other, one of the loop primer and the probe is modified with a fluorescent molecule around the 5' end in the case of the loop primer or the 3' end in the case of the probe, and the other is modified with a quenching molecule.