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Studies on Au NRs Biosensor Based on the Mimic Peptide of Mycobacterium Tuberculosis Antigen for Diagnosis of Tuberculosis

Author: SunWenXia
Tutor: YuanShiShan
School: Hunan Normal University
Course: Pathogen Biology
Keywords: Mycobacterium tuberculosis Mimetic peptides Localized surface plasmon resonance Biosensor Diagnosis
CLC: R52
Type: Master's thesis
Year: 2011
Downloads: 42
Quote: 0
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Abstract


Objective: The gold nanoparticle biosensor is unlabeled biosensor based on localized surface plasmon resonance (Localized surface plasmon resonance, LSPR). Numerous studies confirmed LSPR biosensor can assist in the diagnosis of diseases. In this paper, phage random peptide library screening Mycobacterium tuberculosis antigen mimetic peptide constructed based on the Mycobacterium tuberculosis antigen mimicry peptides gold nanoparticle biosensor to evaluate their ability to diagnose tuberculosis, TB (Tuberculosis, TB) of diagnostic new The theoretical basis and technical support. Methods: ① tuberculosis serum IgG target molecules, three panning phage random peptide library screening with a TB patient serum IgG affinity phage clones, ELISA (Enzyme linked immunosorbent assay, ELISA) identified positive clones; extract positive phage clones ssDNA, sequencing analysis of Mycobacterium tuberculosis antigen analogue peptide sequence; (2) to select high frequency appears analog peptide synthetic ELISA analysis of synthetic peptides corresponding to the positive the sensitivity and specificity of the diagnosis of tuberculosis phage clones; ③ The seed was prepared the gold nanorods base after eleven mercapto undecanoic acid (MUA), 1 - ethyl - (3 - dimethyl-amino-propyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS), chemical modification, the synthesis of short peptide coated gold nanorod constructed biosensor of the gold nanoparticles, to evaluate its ability to diagnose tuberculosis. Results: (1) obtained through the 3 panning phage effective enrichment of the 12 positive phage clones were sequenced to obtain six kinds of short peptide sequence, which has a peptide every four of eight clones each encoding the same; protein BLAST alignment is not found to have homology to the consensus sequence. (2) synthetic high-frequency two 12 peptide (T1 and T2), preliminary identification by ELISA, the sensitivity and specificity were 66.1%, 35.7% and 100.0%, 100.0%. (3) Seed Growth to prepare good dispersity of gold nanorods T1 packet to the gold nanorods, serological testing of 112 serum samples, the sensitivity and specificity of 57.1% and 83.9%, respectively, successfully constructed the diagnostic value of gold nanoparticle biosensors. Conclusions: (1) TB patients serum IgG phage random peptide library to obtain a highly pro-and anti-TB antibody phage display short peptides; (2) seed Prepared a good degree of dispersion of gold nanorods; (3) based on synthetic peptide nano gold biosensors TB has a better diagnostic value.

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CLC: > Medicine, health > Internal Medicine > TB
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