Abstract: Disclosed are a primer probe for PCR detection, a primer probe set, and an application thereof. The primer probe and the primer probe set can simultaneously perform analysis in two dimensions of a fluorescent channel and a melting temperature in a single tube reaction. That is, by using a same fluorescent channel, different targets may be detected by means of melting temperature characteristics; or when different fluorescent channels are used, a target type detection of a product of the number of fluorescent channels and the melting temperature characteristic may be achieved. Moreover, a method for performing multiplex PCR detection by using the primer probe has the advantages of being low in fluorescence background, adjustable in melting temperature, good in inclusiveness, and low in cost, achieving single tube reaction, and being simple and convenient to operate, not easy to cause pollution, high in sensitivity, and wide in applicable range.

Abstract: Disclosed in the present invention are a multiplex nucleic acid detection method, a composition and a kit. The method comprises a multiplex nucleic acid detection method, which comprises the following steps: mixing a composition containing a first primer set and a first probe with a sample from a subject; amplifying a target sequence that may be present in the sample; performing a melting curve analysis to an amplified product in a corresponding detection channel; and judging whether one or more targets are present, according to a result of the melting curve analysis.

Abstract: Specific, accurate, and cost effective primers for performing digital PCR, compositions and kits containing the primer and methods for using and making the same are useful for detecting nucleic acid mutations. A primer useful as a first forward primer in performing digital PCR to detect a target nucleic acid in a sample, includes: a detection portion located upstream to a target sequence binding portion, and including a second forward primer binding portion having a sequence substantially complementary to a second forward primer, and a probe binding portion downstream to the second forward primer binding portion having a sequence substantially complementary to a probe; the target sequence binding portion includes a mismatch portion having a sequence not complementary to the target nucleic acid, and an amplification determinant portion downstream to the mismatch portion having a sequence complementary to a gene allele or a variant thereof encoded by the target nucleic acid.

Abstract: Specific, accurate, and cost effective primers for performing digital PCR, compositions and kits containing the primer and methods for using and making the same are useful for detecting nucleic acid mutations. A primer useful as a first forward primer in performing digital PCR to detect a target nucleic acid in a sample, includes: a detection portion located upstream to a target sequence binding portion, and including a second forward primer binding portion having a sequence substantially complementary to a second forward primer, and a probe binding portion downstream to the second forward primer binding portion having a sequence substantially complementary to a probe; the target sequence binding portion includes a mismatch portion having a sequence not complementary to the target nucleic acid, and an amplification determinant portion downstream to the mismatch portion having a sequence complementary to a gene allele or a variant thereof encoded by the target nucleic acid.