One or more embodiments of the present invention are intended to dissolve the problem of lowering in reaction efficiency of the nucleic acid amplification reaction using a primer with a polynucleotide tag to prepare an amplified product of a target nucleic acid that can be detected on a solid-phase support. One or more other embodiments of the present invention relate to a set of primers comprising: the first primer comprising the first polynucleotide comprising, at the 3′ terminus, polynucleotide A capable of hybridizing to a complementary strand of partial polynucleotide A′ at the 5′ terminus of the target nucleic acid and the first polynucleotide tag, which is a polynucleotide independent of the nucleic acid amplification reaction; the second primer comprising the second polynucleotide comprising, at the 3′ terminus, polynucleotide B capable of hybridizing to partial polynucleotide B′ at the 3′ terminus of the target nucleic acid; and the third primer comprising the third polynucleotide hybridizing to the complementary strand of the target nucleic acid competitively with the polynucleotide A of the first primer but comprising no polynucleotide independent of the nucleic acid amplification reaction.

One or more embodiments of the present invention are intended to dissolve the problem of lowering in reaction efficiency of the nucleic acid amplification reaction using a primer with a polynucleotide tag to prepare an amplified product of a target nucleic acid that can be detected on a solid-phase support. One or more other embodiments of the present invention relate to a set of primers comprising: the first primer comprising the first polynucleotide comprising, at the 3′ terminus, polynucleotide A capable of hybridizing to a complementary strand of partial polynucleotide A′ at the 5′ terminus of the target nucleic acid and the first polynucleotide tag, which is a polynucleotide independent of the nucleic acid amplification reaction; the second primer comprising the second polynucleotide comprising, at the 3′ terminus, polynucleotide B capable of hybridizing to partial polynucleotide B′ at the 3′ terminus of the target nucleic acid; and the third primer comprising the third polynucleotide hybridizing to the complementary strand of the target nucleic acid competitively with the polynucleotide A of the first primer but comprising no polynucleotide independent of the nucleic acid amplification reaction.

An object is to provide a method capable of associating position information of a cell with a cell constituent at a single-cell resolution. That is, the present technology provides an analysis method including: an imaging step of imaging a specimen in a state in which the specimen is being overlapped with a surface to which a molecule having a linker cleavable by stimulation, a barcode sequence, and a target capture portion is immobilized via the linker; an association step of associating a position of a cell with the barcode sequence of the molecule at the position by using a specimen image obtained by the imaging; a cleavage step of selectively stimulating the position of the cell to cleave the linker of the molecule at the position; and a binding step of binding the molecule released from the surface by the cleavage with a constituent of the cell via the target capture portion of the molecule.

An object is to provide a method capable of associating position information of a cell with a cell constituent at a single-cell resolution. That is, the present technology provides an analysis method including: an imaging step of imaging a specimen in a state in which the specimen is being overlapped with a surface to which a molecule having a linker cleavable by stimulation, a barcode sequence, and a target capture portion is immobilized via the linker; an association step of associating a position of a cell with the barcode sequence of the molecule at the position by using a specimen image obtained by the imaging; a cleavage step of selectively stimulating the position of the cell to cleave the linker of the molecule at the position; and a binding step of binding the molecule released from the surface by the cleavage with a constituent of the cell via the target capture portion of the molecule.

The present invention relates to methods for the detection of a sexually transmitted infectious pathogens in human subjects. The detection can be done without sample purification and on several types of pathogens. The methods are ideal for urine samples. One pathogen is Chlamydia trachomatis.

The present invention provides microfluidic porous substrate-based devices for multiplexed biosensing. The devices are suitable for detecting viruses and bacteria, such as by way of detecting pathogenic genes and antibodies. The devices support reverse transcriptase loop-mediated isothermal amplification for rapid results within minutes. The devices also support vertical-lateral-vertical direction flow assays, such as in the form of a multi-layered adhesive bandage.

The present invention provides microfluidic porous substrate-based devices for multiplexed biosensing. The devices are suitable for detecting viruses and bacteria, such as by way of detecting pathogenic genes and antibodies. The devices support reverse transcriptase loop-mediated isothermal amplification for rapid results within minutes. The devices also support vertical-lateral-vertical direction flow assays, such as in the form of a multi-layered adhesive bandage.

An object of the invention is to provide an optical detection method and a quantification method of a product obtained by amplifying a target nucleic acid contained in a paper piece of a blood spot in a filter paper obtained by blotting blood on a filter paper and then drying the blood using nucleic acid amplification reaction by real-time PCR and to further provide a kit used for the methods. The invention provides a method and a kit which can optically detect and quantify a target nucleic acid from a dried blood spot in a filter paper by real-time PCR without any complicated pretreatment, by adjusting the size of the punched piece of the dried blood spot in a filter paper or the whole blood amount contained in the punched piece, the amount of the PCR reaction reagent, or performing the PCR reaction in a PCR reaction tube closed with a cap.

An object of the invention is to provide an optical detection method and a quantification method of a product obtained by amplifying a target nucleic acid contained in a paper piece of a blood spot in a filter paper obtained by blotting blood on a filter paper and then drying the blood using nucleic acid amplification reaction by real-time PCR and to further provide a kit used for the methods. The invention provides a method and a kit which can optically detect and quantify a target nucleic acid from a dried blood spot in a filter paper by real-time PCR without any complicated pretreatment, by adjusting the size of the punched piece of the dried blood spot in a filter paper or the whole blood amount contained in the punched piece, the amount of the PCR reaction reagent, or performing the PCR reaction in a PCR reaction tube closed with a cap.

Sensitive and specific detection of nucleic acids can be achieved using a chemical ligation-based template assisted rapid assay (TARA-L) with simple chemical reactions between probes and without the need for enzymes. Probes are designed to form a ligation product when they anneal to adjacent portions of a target nucleic acid. The ligation products can be detected, such as in immunochromatographic assays. The methods allow for the fast, efficient analysis of biological samples for the presence of nucleic acids and can be used, for example, in point of care settings.