Multiplexed, rapid detection of H5N1 using a PCR-free nanoparticle-based genomic microarray assay.
Multiplexed, rapid detection of H5N1 using a PCR-free nanoparticle-based genomic microarray assay.
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DOI：
10.1186/1472-6750-10-74
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发表时间：
2010-10-13
影响因子：
3.5
通讯作者：
Hewlett IK
中科院分区：
文献类型：
--
作者：
Zhao J;Tang S;Storhoff J;Marla S;Bao YP;Wang X;Wong EY;Ragupathy V;Ye Z;Hewlett IK
十多年来，人们越来越关注将纳米技术和微阵列平台用于诊断应用。在这份报告中，我们描述了一种快速和简单的金纳米颗粒（NP）为基础的基因组芯片检测禽流感病毒H5 N1的特异性鉴定和其他主要的流感病毒株（H1N1，H3 N2）的歧视。基于H1N1、H3 N2和H5 N1病毒的基质（M）基因的共有序列以及H5 N1病毒的血凝素（HA）和神经氨酸酶（NA）基因的特异性序列设计捕获和中间寡核苷酸。在不存在RNA片段化、靶扩增和酶促反应的情况下，使用捕获-靶-中间寡核苷酸杂交和金NP介导的银染色在2.5小时内检测到病毒RNA。该检测方法的检测下限（LOD）是小于100 fM的纯化的PCR片段和103 TCID 50单位的H5 N1病毒RNA。基于NP的微阵列分析能够检测和区分H5 N1序列与主要的甲型流感病毒（H1N1，H3 N2）。这里描述的新方法可能是有用的同时检测和亚型的主要流感病毒A。
For more than a decade there has been increasing interest in the use of nanotechnology and microarray platforms for diagnostic applications. In this report, we describe a rapid and simple gold nanoparticle (NP)-based genomic microarray assay for specific identification of avian influenza virus H5N1 and its discrimination from other major influenza A virus strains (H1N1, H3N2). Capture and intermediate oligonucleotides were designed based on the consensus sequences of the matrix (M) gene of H1N1, H3N2 and H5N1 viruses, and sequences specific for the hemaglutinin (HA) and neuraminidase (NA) genes of the H5N1 virus. Viral RNA was detected within 2.5 hours using capture-target-intermediate oligonucleotide hybridization and gold NP-mediated silver staining in the absence of RNA fragmentation, target amplification, and enzymatic reactions. The lower limit of detection (LOD) of the assay was less than 100 fM for purified PCR fragments and 103 TCID50 units for H5N1 viral RNA. The NP-based microarray assay was able to detect and distinguish H5N1 sequences from those of major influenza A viruses (H1N1, H3N2). The new method described here may be useful for simultaneous detection and subtyping of major influenza A viruses.
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影响因子：
9.4
作者：
Spackman, E;Senne, DA;Suarez, DL
通讯作者：
Suarez, DL
影响因子：
9.4
作者：
Mehlmann, Martin;Dawson, Erica D.;Rowlen, Kathy L.
通讯作者：
Rowlen, Kathy L.
影响因子：
9.4
作者：
Fouchier, RAM;Bestebroer, TM;Osterhaus, ADME
通讯作者：
Osterhaus, ADME
DOI：
10.1073/pnas.78.7.4170
发表时间：
1981-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
LAMB, RA;LAI, CJ;CHOPPIN, PW
通讯作者：
CHOPPIN, PW
影响因子：
9.4
作者：
Kessler, N;Ferraris, O;Steel, A
通讯作者：
Steel, A