Strengthening of alkaline phosphatase by SDSds as a chemical luminescent substrate and enzyme inhibition process

摘要

The invention relates to a method of detecting a target oligonucleotide in a sample, comprising: (a) providing a support-bound capture probe (CP) comprising a region having a nucleic acid sequence C-3; (b) providing a label probe (LP) comprising a region having a nucleic acid sequence L-3, wherein the label probe contains a label moiety that is capable of generating a detectable signal; (c) providing a label extender (LE) comprising a region having first and second nucleic acid sequences, wherein the first LE nucleic acid sequences L-2 is complementary to nucleic acid sequences L-3 and the second LE nucleic acid sequence L-1 is complementary to a nucleic acid sequence in the target analyte; (d) providing a capture extender (CE) comprising a region having first and second nucleic acid sequences, wherein the first CE nucleic acid sequences C-2 is complementary to nucleic acid sequence C-3 and the second CE nucleic acid sequence C-1 is complementary to the second LE nucleic acid sequence L-1; (e) incubating the sample with the LE and the LP under first hybridizing conditions to form a first reaction mixture containing an LP/LE/target hybrid complex and an unbound LP/LE hybrid complex; (f) incubating the first reaction mixture with a solid support surface containing a surface-bound CE/CP hybrid complex under second hybridizing conditions, thereby forming a second reaction mixture containing a surface-bound LP/LE/target/CE/CP hybrid complex and an unbound LP/LE/target complex; (g) thereafter separating materials not bound to the solid support from those bound to the solid support; and (h) detecting the presence of label in the support-bound, LP/LE/target/CE/CP hybrid complex.