Assay for the parallel detection of biological material based on PCR

A forward primer and reverse primer technology, applied in the field of proteins, peptides, lipids or carbohydrates in the detection sample, which can solve the problems of low sensitivity and lack of reproducibility

Inactive Publication Date: 2015-07-29
NAT UNIV OF SINGAPORE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although high-throughput peptide-based microarrays for antibody monitoring have been reported, they have not been widely used due to low sensitivity and lack of reproducibility (H. Andresen and C. (2009) Deciphering the antibodyome-peptide arrays for serum antibody biomarker diagnostics. Current Proteomics, 2009, 6, 1-12)

Method used

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  • Assay for the parallel detection of biological material based on PCR
  • Assay for the parallel detection of biological material based on PCR
  • Assay for the parallel detection of biological material based on PCR

Examples

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Embodiment Construction

[0104] The feasibility of the invention is demonstrated herein using two exemplary probes / P-O and positive antibodies specific for each. The target proteins in this example are epitopes specific for influenza virus and dengue virus.

[0105] P-O conjugate and primer design (see Figure 5 ):

[0106] 1) Select two specific peptide epitopes (YPYDVPDYA and YKQPLWPNQISW, in Figure 5 Shown on the left hand side of part A): one from influenza virus and the other from dengue virus.

[0107] 2) In this particular example, specific restriction sites are incorporated into each of the different barcode regions for oligonucleotide design, i.e., different restriction sites are incorporated into each barcode region so that Use enzymatic digestion to confirm sequencing results. BamH1 was incorporated into the barcode region of the influenza-specific probe, whereas HindIII was incorporated into the barcode region of the dengue-specific probe.

[0108] 3) In this particular example, an 8...

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Abstract

The invention concerns a novel parallel method for detecting biological material, in particular peptides or proteins, in a sample at least one probe for use in the said method, a plurality, or library, of said probes for use in said method, and a kit of parts for carrying out said method wherein said probe comprises a binding partner that is specific for said peptide or protein and, attached thereto, an oligonucleotide comprising: i) a first sequence that is complementary to a forward primer sequence for amplification of said oligonucleotide; ii) a second sequence that is complementary to a reverse primer sequence for amplification of said oligonucleotide; and iii) positioned between said first and second sequences an identification sequence of nucleotides or barcode.

Description

[0001] The present invention relates to novel methods for detecting at least one target molecule in a sample, in particular for detecting peptides, proteins, lipids or carbohydrates in a sample; at least one probe for said method; for said method A plurality of such probes or a library of such probes, and kits for carrying out the methods. Background technique [0002] Molecular interactions involving e.g. peptides or proteins (e.g., antibody-antigen interactions, hormone-receptor interactions, virus-receptor interactions, enzyme-substrate interactions, etc.) represent the most complex and important interactions in any biological system. the process of. Its detection can provide valuable information about the state of the system and thus can provide important information of diagnostic, therapeutic or commercial value. For example, the following is a non-exhaustive list of the types of information that can be obtained by monitoring peptide or protein interactions. [0003] a)...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/53C07K19/00
CPCC12Q1/686C07K19/00G01N33/531G01N33/58G01N33/537C12Q1/6844G01N33/53C12Q2537/143C12Q2563/185C12Q1/6876C12Q2600/16
Inventor 王林发黄英勇奥克托布尔·迈克尔·塞申斯达尼埃尔·伊丽莎白·安德森
Owner NAT UNIV OF SINGAPORE
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