title: Closed-Tube PCR with Nested Serial Invasion Probe Visualization Using Gold Nanoparticles.
abstract: Detecting DNA biomarkers related to personalized medicine could improve the outcome of drug therapy. However, personalized medicine in a resource-restrained hospital is very difficult because DNA biomarker detection should be performed by well-trained staff and requires expensive laboratory facilities.
  
  
  
  We developed a gold nanoparticle-based "Tube-Lab" to enable DNA analysis in a closed tube. Gold nanoparticle-modified probes (GNPs) were used to construct an inexpensive and simple DNA sensor for signal readout. The method consists of 3 steps (template amplification, sequence identification, and GNP-based signal readout), bridged by an invasive reaction. With temperature control at each step, the 3 reactions proceed sequentially and automatically in a closed tube without any liquid transfer. We used Tube-Lab to detect different biomarkers in blood, tissue, and plasma, including US Food and Drug Administration-approved pharmacogenomic biomarkers (single nucleotide polymorphisms, somatic mutations).
  
  
  
  The combination of PCR-based template replication and invader-based signal amplification allowed detection of approximately 6 copies of input DNA and the selective pick up 0.1% mutants from large amounts of background DNA. This method highly discriminated polymorphisms and somatic mutations from clinical samples and allowed a "liquid biopsy" assay with the naked eye.
  
  
  
  Tube-Lab provides a promising and cost-effective approach for DNA biomarker analysis, including polymorphisms and somatic mutations from blood DNA, tissue DNA, or circulating tumor DNA in plasma, which are critical for personalized medicine.
authors.affiliation: Department of Pharmacology, Jinling Hospital, State Key Laboratory of Analytical Chemistry for Life Science, Medical School of Nanjing University, Nanjing, China.|Guangzhou Biotron Technology Co. Ltd., Guangzhou, China.
authors.forename: Jianping
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Pharmacology, Jinling Hospital, State Key Laboratory of Analytical Chemistry for Life Science, Medical School of Nanjing University, Nanjing, China.
authors.forename: Bingjie
authors.initials: B
authors.lastname: Zou
authors.affiliation: Department of Pharmacology, Jinling Hospital, State Key Laboratory of Analytical Chemistry for Life Science, Medical School of Nanjing University, Nanjing, China.
authors.forename: Yinjiao
authors.initials: Y
authors.lastname: Ma
authors.affiliation: Department of Pharmacology, Jinling Hospital, State Key Laboratory of Analytical Chemistry for Life Science, Medical School of Nanjing University, Nanjing, China.
authors.forename: Xueping
authors.initials: X
authors.lastname: Ma
authors.affiliation: Department of Pharmacology, Jinling Hospital, State Key Laboratory of Analytical Chemistry for Life Science, Medical School of Nanjing University, Nanjing, China.
authors.forename: Nan
authors.initials: N
authors.lastname: Sheng
authors.affiliation: Department of Pharmacology, Jinling Hospital, State Key Laboratory of Analytical Chemistry for Life Science, Medical School of Nanjing University, Nanjing, China.
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Rui
authors.affiliation: GENESEEQ Biotechnology Inc., Nanjing, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Shao
authors.affiliation: Department of Pharmacology, Jinling Hospital, State Key Laboratory of Analytical Chemistry for Life Science, Medical School of Nanjing University, Nanjing, China; ghzhou@nju.edu.cn.
authors.forename: Guohua
authors.initials: G
authors.lastname: Zhou
mesh_terms: D015415:Biomarkers / Q000097:blood; D004247:DNA / Q000097:blood / Q000235:genetics*; D015342:DNA Probes / Q000097:blood* / Q000235:genetics; D006046:Gold / Q000737:chemistry*; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry*; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*
chemical_list: D015415:Biomarkers; D015342:DNA Probes; D006046:Gold; D004247:DNA