An integrated microfluidic device utilizing vancomycin conjugated magnetic beads and nanogold-labeled specific nucleotide probes for rapid pathogen diagnosis

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• 作者：Chih-Hung Wang ; PhD^a ; ^{biowang2007@gmail.com" class="auth_mail} ; Chia-Jung Chang^a ; ^{jerry780620@gmail.com" class="auth_mail} ; Jiunn-Jong Wu ; PhD^b ; ^{jjwu@mail.ncku.edu.tw" class="auth_mail} ; Gwo-Bin Lee ; PhD^a ; ^c ; ^d ; ^{gwobin@pme.nthu.edu.tw" class="auth_mail}
• 关键词：Bp ; base-pair ; CFU ; colony forming unit ; DNA ; deoxyribonucleic acid ; dNTP ; deoxyribonucleotide triphosphate ; E. coli ; Escherichia coli ; EDAC ; (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide ; EMVs ; electromagnetic valves ; ddH2O ; deionic distilled water ; G(&minus ; ) ; Gram-negative bacteria ; G(+) ; Gram-positive bacteria ; HCl ; hydrogen chloride ; LB ; Luria-Betani ; LOC ; lab-on-a-chip ; LOD ; limit of detection ; MEMS ; micro-electro-mechanical-systems ; MRSA ; methicillin-resistant Staphylococcus aureus ; PCR
• 刊名：Nanomedicine: Nanotechnology, Biology and Medicine
• 出版年：May 2014
• 年：2014
• 卷：10
• 期：4
• 页码：809-818
• 全文大小：833 K

文摘

A PCR-free assay for rapid pathogen diagnosis was implemented on an integrated microfluidic system in this study. Vancomycin-conjugated magnetic beads were used to capture multiple strains of bacteria and nucleotide probes labeled gold nanoparticles were used to specify and detect a specific strain by hybridization-induced color change. The assay was entirely automated within an integrated microfluidic device that was composed of suction-type micropumps, microvalves, microchannels, and microchambers that fabricated by microfluidic technology. Multiple strains of bacteria could be captured simultaneously by vancomycin-conjugated magnetic beads, with capturing efficiency exceeding 80%. Subsequently, sensitive and strain-specific detection against target bacteria could be achieved by using nanogold labeled specific nucleotide probes. The limit of detection of 10² CFU bacteria was achieved. Importantly, nucleic acid amplification was not involved in the diagnostic procedures; the entire analytic process required only 25 min. The developed platform may provide a promising tool for rapid diagnosis of bacterial infections.

From the Clinical Editor

In this novel study, a PCR-free pathogen detection method is demonstrated. After vancomycin-conjugated magnetic beads captured bacteria, nucleotide probes-labeled gold nanoparticles were employed to specify and detect specific strains via hybridization-induced color change. Multiple strains of bacteria could be captured simultaneously with an efficiency exceeding 80%, enabling the detection of as low as 10² CFU of bacteria.