Methods according to aspects of the disclosure, compositions and kits therefore, include at least one, two, or three sets of amplification primers and hydrolysis probes with at least two separate corresponding readouts per set. According to aspects of the present disclosure, the at least two hydrolysis probes and associated pair of primers of each set are directed to opposite strands of an amplification product of the set. According to aspects of the present disclosure, one of the two hydrolysis probes in each set is directed to a first strand of the amplification product and therefore has a sequence complementary to the first strand of the amplification product and the second of the two hydrolysis probes in the set is directed to the second strand of the amplification product and therefore has a sequence complementary to the second strand of the amplification product.

Methods according to aspects of the disclosure, compositions and kits therefore, include at least one, two, or three sets of amplification primers and hydrolysis probes with at least two separate corresponding readouts per set. According to aspects of the present disclosure, the at least two hydrolysis probes and associated pair of primers of each set are directed to opposite strands of an amplification product of the set. According to aspects of the present disclosure, one of the two hydrolysis probes in each set is directed to a first strand of the amplification product and therefore has a sequence complementary to the first strand of the amplification product and the second of the two hydrolysis probes in the set is directed to the second strand of the amplification product and therefore has a sequence complementary to the second strand of the amplification product.

Methods for the early detection of subjects at risk of developing rheumatoid arthritis, and subjects having early rheumatoid arthritis thus allowing for early intervention.

Methods for the early detection of subjects at risk of developing rheumatoid arthritis, and subjects having early rheumatoid arthritis thus allowing for early intervention.

A microfluidic device for amplifying a nucleic acid includes a cartridge and a control part. The cartridge includes a tank part and a plurality of first chambers. The control part is configured to control execution of a thermal cycle, count a number of repetitions of the thermal cycle for each of the first chambers and store a count value, acquire a fluorescence intensity of each of the first chambers for each thermal cycle, and reset the count value of a defective chamber of which the fluorescence intensity is not within a predetermined range, discharge the solution from the defective chamber, and fill the defective chamber with a new solution from the tank part.

A microfluidic device for amplifying a nucleic acid includes a cartridge and a control part. The cartridge includes a tank part and a plurality of first chambers. The control part is configured to control execution of a thermal cycle, count a number of repetitions of the thermal cycle for each of the first chambers and store a count value, acquire a fluorescence intensity of each of the first chambers for each thermal cycle, and reset the count value of a defective chamber of which the fluorescence intensity is not within a predetermined range, discharge the solution from the defective chamber, and fill the defective chamber with a new solution from the tank part.

Provided herein are methods of treating a patient afflicted with a tumor according to the tumor's microenvironments (TME). Also provided are gene panels that can be used for identifying a human subject afflicted with a cancer suitable for treatment with a particular therapeutic agent based on the subject's TME.