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US 2023/0144631 A1Published application
Universal probe chip-based multiplex quantitative pcr testing system
AssigneeFlashdx Shenzhen Inc.
InventorMin YUE
Life of this patent
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Abstract
Provided is a surface probe-based quantitative PCR testing system, comprising: (a) a solid phase carrier; (b) a specific primer pair of a sequence to be tested, which comprises a first primer and a second primer; and (c) a quenching probe. Further provided are a method for quantitative PCR testing and a kit.
Claims10
A surface probe-based quantitative PCR detection system, the detection system comprising.
- (a) a solid support, the solid support having n sub-detection zones on a main surface, wherein n is a positive integer of ≥ 2 and at least one of the sub-detection zones is a surface quantification sub-detection zone;
- wherein each of the surface quantification sub-detection zone is independently immobilized with a microarray surface probe, the microarray surface probe being a single stranded nucleic acid, and one end of the microarray surface probe is immobilized on the surface of the solid support, and the microarray surface probe carries a first detectable marker, the first detectable marker being selected from the group consisting of:
- a fluorescent group, luminescent group, luminescent marker, quantum dot, and a combination thereof;
- (b) a primer pair specific to the sequence to be detected, comprising a first primer and a second primer;
- (c) a quenching probe, a quencher is attached to one side or one end or in the middle of the quenching probe;
- and wherein the quenching probe and at least one microarray surface probes of the surface quantification sub-detection zone can be combined to form a double-stranded structure, and in the double-stranded structure, the quenching group of the quenching probe causes the signal of the first detectable marker (e.g. fluorescent group) of the microarray surface probe to be quenched in whole or in part;
- when the concentration of the quenching probe in the detection system is reduced, the degree of quenching of the signal of the first detectable marker (e.g. fluorescent group) of the microarray surface probe of at least one surface quantification sub-detection zone is reduced.
4 more elements
A kit for a quantitative PCR detection comprising:
- (a) a first container and a solid support located in the first container ,
- the solid support having n sub-detection zones on a main surface, wherein n is a positive integer of 2 and at least one of the sub-detection zones is a surface quantification sub-detection zone;
- wherein each of the surface quantification sub-detection zone is independently immobilized with a microarray surface probe, the microarray surface probe being a single stranded nucleic acid, and one end of the microarray surface probe is immobilized on the surface of the solid support, and the microarray surface probe carries a first detectable marker, the first detectable marker being selected from the group consisting of:
- a fluorescent group, luminescent group, luminescent marker, quantum dot, and a combination thereof;
- (b) a second container and (b1) a primer pair specific to the sequence to be detected, comprising a first primer, a second primer;
- and (b2) a quenching probe, a quencher is attached to one side or one end or in the middle of the quenching probe, located in the second container;
- and wherein the quenching probe and at least one microarray surface probes of the surface quantification sub-detection zone can be combined to form a double-stranded structure, and in the double-stranded structure, the quenching group of the quenching probe causes the signal of the first detectable marker (e.g. fluorescent group) of the microarray surface probe to be quenched in whole or in part;
- when the concentration of the quenching probe in the detection system is reduced, the degree of quenching of the signal of the first detectable marker (e.g. fluorescent group) of the microarray surface probe of at least one surface quantification sub-detection zone is reduced;
- (c) optionally a third container and a buffer or buffer component for PCR amplification located in the third container;
- (d) optionally a fourth container and a polymerase for PCR amplification located in the fourth container;
- (e) optionally a fifth container and a universal amplification primer pair located in the fifth container;
- and
- (f) optionally a specification, the specification describes a method for performing a quantitative PCR detection.
10 more elements
Dependencies are inferred from the claim text.