ICP-MS-Based Multiplex and Ultrasensitive Assay of Viruses with Lanthanide-Coded Biospecific Tagging and Amplification Strategies

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• 作者：Yacui Luo ; Xiaowen Yan ; Yishun Huang ; Ruibin Wen ; Zhaoxin Li ; Limin Yang ; Chaoyong James Yang ; Qiuquan Wang
• 刊名：Analytical Chemistry
• 出版年：2013
• 出版时间：October 15, 2013
• 年：2013
• 卷：85
• 期：20
• 页码：9428-9432
• 全文大小：296K
• 年卷期：v.85,no.20(October 15, 2013)
• ISSN：1520-6882

文摘

Highly sensitive and a multiplex assay of viruses and viral DNAs in complex biological samples is extremely important for clinical diagnosis and prognosis of pathogenic diseases as well as virology studies. We present an effective ICP-MS-based multiplex and ultrasensitive assay of viral DNAs with lanthanide-coded oligonucleotide hybridization and rolling circle amplification (RCA) strategies on biofunctional magnetic nanoparticles (MNPs), in which single-stranded capture DNA (ss-Cap-DNA)-functionalized MNPs (up to 1.65 脳 10⁴ ss-Cap-DNA per MNP) were used to recognize and enrich target DNAs, and single-stranded report DNA (ss-Rep-DNA-DOTA-Ln) coded by the lanthanide鈥揇OTA complex hybridized with the targeted DNA for highly sensitive readout of HIV (28 amol), HAV (48 amol), and HBV (19 amol). When utilizing the RCA technique in association with the design and synthesis of a 鈥渂ridge鈥?DNA and a corresponding ss-Rep-DNA-DOTA-Ho, as low as 90 zmol HBV could be detected. Preliminary applications to the determination of the viral DNAs in 4T1 cell lysates and in serum confirmed the feasibility of this ICP-MS-based multiplex DNA assay for clinical use. One can expect that this element-coded ICP-MS-based multiplex and ultrasensitive DNA assay will play an ever more important role in the fields of bioanalysis and virology and in medical studies after further sophisticated modifications.