Shiping Song

Publications (67)440.17 Total impact

• Article: Diagnosis of schistosomiasis japonica with interfacial co-assembly-based multi-channel electrochemical immunosensor arrays.

  Wangping Deng, Bin Xu, Haiyan Hu, Jianyong Li, Wei Hu, Shiping Song, Zheng Feng, Chunhai Fan
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  ABSTRACT: Schistosomiasis control remains to be an important and challenging task in the world. However, lack of quick, simple, sensitive and specific sero-diagnostic test is still a hurdle in the control practice. The commonly employed enzyme-linked immuno-sorbent assay (ELISA) relies on the native soluble egg antigen (SEA) that is limited in supply. Here we developed an electrochemical immunosensor array (ECISA) assay with an interfacial co-assembly strategy. A recombinant Schistosoma japonicum (Sj) calcium-binding protein (SjE16) was used as a principal antigen, while the SEA as a minor, co-assembling agent, with a ratio of 8:1 (SjE16: SEA, Sj16EA), which was co-immobilized on a disposable 16-channel screen-printed carbon electrode array. A portable electrochemical detector was employed to detect antibodies in serum samples. The sensitivity of ECISA reached 100% with minimal cross-reactions. Therefore, we have demonstrated that this rapid, sensitive and specific ECISA technique has the potential to perform large-scale on-site screening of Sj infection.
  Scientific Reports 05/2013; 3:1789.
• Article: Rolling Circle Amplification-Based DNA Origami Nanostructrures for Intracellular Delivery of Immunostimulatory Drugs.

  Xiangyuan Ouyang, Jiang Li, Huajie Liu, Bin Zhao, Juan Yan, Yinzhou Ma, Shoujun Xiao, Shiping Song, Qing Huang, Jie Chao, Chunhai Fan
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  ABSTRACT: Several single-stranded scaffold DNA, obtained from rolling circle amplification (RCA), are folded by different staples to form DNA nanoribbons. These DNA nanoribbons are rigid, simple to design, and cost-effective drug carriers, which are readily internalized by mammalian cells and show enhanced immunostimulatory activity.
  Small 04/2013; · 8.35 Impact Factor
• Article: Carbon Nanotubes Multifunctionalized by Rolling Circle Amplification and Their Application for Highly Sensitive Detection of Cancer Markers.

  Bin Zhao, Juan Yan, Dongfang Wang, Zhilei Ge, Shijiang He, Dannong He, Shiping Song, Chunhai Fan
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  ABSTRACT: There are still challenges for the development of multifunctional carbon nanotubes (CNTs). Here, a multiwalled carbon nanotube (MWCNT)-based rolling circle amplification system (CRCAS) is reported which allows in situ rolling circle replication of DNA primer on the surface of MWCNTs to create a long single-strand DNA (ssDNA) where a large number of nanoparticles or proteins could be loaded, forming a nano-biohybridized 3D structure with a powerful signal amplification ability. In this strategy, the binding ability of proteins, hybridization, replication ability of DNA, and the catalytical ability of enzymes are integrated on a single carbon nanotube. The CRCAS is then used to develop colorimetric and chemiluminescent assays for the highly sensitive and specific detection of cancer protein markers, alpha-fetoprotein (AFP) and prostate specific antigen (PSA). The colorimetric CRCAS assay is 4000 times more sensitive than a conventional enzyme-linked immunosorbent assay (ELISA), and its concentration range is 10 000 times wider. Control experiments show that as low as 10 pg mL(-1) AFP or PSA could be detected even in the presence of interfering protein markers with a more than 10(5) -fold greater concentration in the sample, demonstrating the high specificity of the CRCAS assay. The limit of detection of the chemiluminescent CRCAS assays for AFP and PSA are 5 fg mL(-1) (70 aM) and 10 fg mL(-1) (0.29 fM), respectively, indicating that the sensitivity is much higher than that of the colorimetric CRCAS assay. Importantly, CRCAS works well with real biological samples.
  Small 03/2013; · 8.35 Impact Factor
• Article: A power-free microfluidic chip for SNP genotyping using graphene oxide and a DNA intercalating dye.

  Jing Li, Yan Huang, Dongfang Wang, Bo Song, Zhenhua Li, Shiping Song, Lihua Wang, Bowei Jiang, Xingchun Zhao, Juan Yan, Rui Liu, Dannong He, Chunhai Fan
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  ABSTRACT: We herein report a power-free microfluidic chip for fluorescent DNA detection with high single-nucleotide polymorphism discrimination, using a DNA intercalator and graphene oxide.
  Chemical Communications 03/2013; · 6.17 Impact Factor
• Article: Single-nucleotide polymorphism genotyping using a novel multiplexed electrochemical biosensor with nonfouling surface.

  Gang Liu, Ruojun Lao, Li Xu, Qin Xu, Lanying Li, Min Zhang, Shiping Song, Chunhai Fan
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  ABSTRACT: A novel electrochemical DNA biosensor for single-nucleotide polymorphism (SNP) analysis was developed. In this work, an oligonucleotide-incorporated nonfouling surface (ONS) was constructed to resist nonspecific absorption. The biosensor was developed using a 16-electrode array for high-throughput SNP analysis. The proposed strategy was primarily based on specific oligonucleotide ligation. Fully matched target DNA templated the ligation between a capture probe assembled on gold electrodes and a tandem signal probe with a biotin moiety that could capture avidin-horseradish peroxidase and sequentially generate a catalysed amperometric signal. A pre-core mutation in the hepatitis B virus (HBV) genome at G1896A and two adjacent polymorphisms in the human CYP2C19 genome at C680T and G681A were analysed. Polymerase chain reaction (PCR) products were used as real-life samples and analysed. Our results showed that 10% of a single-mismatched mutant gene was clearly distinguished with a current signal 16 times higher than that of the blank sample, demonstrating the selectivity and practicability of the multiplexed electrochemical DNA biosensor.
  Biosensors & bioelectronics 11/2012; 42C:516-521. · 5.43 Impact Factor
• Article: Nano Rolling-Circle Amplification for Enhanced SERS Hot Spots in Protein Microarray Analysis.

  Juan Yan, Shao Su, Shijiang He, Yao He, Bin Zhao, Dongfang Wang, Honglu Zhang, Qing Huang, Shiping Song, Chunhai Fan
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  ABSTRACT: Although "hot spots" have been proved to contribute to surface enhanced Raman scattering (SERS), less attention was paid to increase the number of the "hot spot" to directly enhance the Raman signals in bioanalytical systems. Here we report a new strategy based on nano rolling-circle amplification (nanoRCA) and nano hyperbranched rolling-circle amplification (nanoHRCA) to increase "hot spot" groups for protein microarrays. First, protein and ssDNA are coassembled on gold nanoparticles, making the assembled probe have both binding ability and hybridization ability. Second, the ssDNAs act as primers to initiate in situ RCA reaction to produced long ssDNAs. Third, a large number of SERS probes are loaded on the long ssDNA templetes, allowing thousands of SERS probes involved in each biomolecular recognition event. The strategy offered high-efficiency Raman enhancement and could detect less than 10 zeptomolar protein molecules in protein microarray analysis.
  Analytical Chemistry 10/2012; · 5.86 Impact Factor
• Article: A surface-initiated enzymatic polymerization strategy for electrochemical DNA sensors.

  Ying Wan, Hui Xu, Yan Su, Xinhua Zhu, Shiping Song, Chunhai Fan
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  ABSTRACT: In this work, we report a novel strategy of electrochemical DNA (E-DNA) sensor based on surface initiated enzymatic polymerization (SIEP). This DNA sensor employs a capture DNA probe labeled with thiol at its 3' terminal to be immobilized at gold electrode via gold-thiol chemistry. Oligo (ethylene glycol) -terminated thiols (SH-OEGs) are then used to prepare an oligonucleotide-incorporated nonfouling surface (ONS). After the sequence-specific recognition of target DNA, terminal deoxynucleotidyl transferase (TdT) is employed to catalyze the sequential addition of deoxynucleotides (dNTPs) at the 3'-OH group of target DNA without template. During the TdT-mediated extension reaction, by using biotinlated 2'-deoxyadenosine 5'-triphosphate (biotin-dATP), biotin labels are incorporated into the SIEP-generated long single-stranded DNA (ssDNA). Specific binding of avidin-horseradish peroxidase (Av-HRP) to the biotin label leads to enzyme turnover-based signal transduction. By using this new strategy, we demonstrated the high picomolar sensitivity and a broad detection range of six orders of magnitude.
  Biosensors & bioelectronics 09/2012; · 5.43 Impact Factor
• Article: Electrochemical single nucleotide polymorphisms genotyping on surface immobilized three-dimensional branched DNA nanostructure

  ZhiLei Ge, Hao Pei, LiHua Wang, ShiPing Song, ChunHai Fan
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  ABSTRACT: An electrochemical assay for single nucleotide polymorphisms (SNPs) genotyping is reported. Although electrochemical method is sensitive for DNA detection on surfaces, the ability of surface assay to precisely recognize DNA hybridization event is sacrificed to some extent due to the crowded confined surfaces environments that disfavor DNA hybridization. In the present study, we employed branched tetrahedron structure probes (TSPs) to replace regular linear single stranded DNA capture probes that were immobilized on solid surfaces. This three-dimensional DNA nanostructure lowers the density of immobilized DNA probes on confined surfaces, providing a hybridization environment that is similar to homogenous solution. This TSP-based electrochemical assay reveals excellent performance for SNPs genotyping with concentration as low as 1 nM. KeywordsDNA nanostructure–surface density–electrochemical
  Science China-Chemistry 05/2012; 54(8):1273-1276. · 1.02 Impact Factor
• Article: Gold nanoparticlebased optical probes for target-responsive DNA structures

  Lihua Wang, Juan Zhang, Xun Wang, Qing Huang, Dun Pan, Shiping Song, Chunhai Fan
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  ABSTRACT: In this work, we report the use of unmodified gold nanoparticles (AuNPs) as an optical probe for the detection of target-responsive structural variations of DNA. By employing two DNA structures, i.e., a pH-responsive i-motif oligonucleotide and a mercury-specific oligonucleotide (MSO), we demonstrated that AuNPs could selectively distinguish target-free and target-bound oligonucleotides via the characteristic surface plasmon resonance-associated red-to-blue color change. Based on these observations, we developed a convenient “mix-and-detect” approach that could selectively detect environmentally toxic mercury ions.
  Gold bulletin 05/2012; 41(1):37-41. · 3.52 Impact Factor
• Source

  Available from: Lihua Wang

  Article: Design of a carbon nanotube/magnetic nanoparticle-based peroxidase-like nanocomplex and its application for highly efficient catalytic oxidation of phenols

  Xiaolei Zuo, Cheng Peng, Qing Huang, Shiping Song, Lihua Wang, Di Li, Chunhai Fan
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  ABSTRACT: We report a novel nanotechnology-based approach for the highly efficient catalytic oxidation of phenols and their removal from wastewater. We use a nanocomplex made of multi-walled carbon nanotubes (MWNTs) and magnetic nanoparticles (MNPs). This nanocomplex retains the magnetic properties of individual MNPs and can be effectively separated under an external magnetic field. More importantly, the formation of the nanocomplex enhances the intrinsic peroxidase-like activity of the MNPs that can catalyze the reduction of hydrogen peroxide (H2O2). Significantly, in the presence of H2O2, this nanocomplex catalyzes the oxidation of phenols with high efficiency, generating insoluble polyaromatic products that can be readily separated from water.
  Nano Research 04/2012; 2(8):617-623. · 6.97 Impact Factor
• Article: Graphene-based high-efficiency surface-enhanced Raman scattering-active platform for sensitive and multiplex DNA detection.

  Shijiang He, Keng-Ku Liu, Shao Su, Juan Yan, Xiuhai Mao, Dongfang Wang, Yao He, Lain-Jong Li, Shiping Song, Chunhai Fan
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  ABSTRACT: We have developed a surface-enhanced Raman scattering (SERS)-active substrate based on gold nanoparticle-decorated chemical vapor deposition (CVD)-growth graphene and used it for multiplexing detection of DNA. Due to the combination of gold nanoparticles and graphene, the Raman signals of dye were dramatically enhanced by this novel substrate. With the gold nanoparticles, DNA capture probes could be easily assembled on the surface of graphene films which have a drawback to directly immobilize DNA. This platform exhibits extraordinarily high sensitivity and excellent specificity for DNA detection. A detection limit as low as 10 pM is obtained. Importantly, two different DNA targets could be detected simultaneously on the same substrate just using one light source.
  Analytical Chemistry 04/2012; 84(10):4622-7. · 5.86 Impact Factor
• Article: The enzyme-amplified amperometric DNA sensor using an electrodeposited polymer redox mediator

  LanYong Zhang, Ying Wan, Jiong Zhang, Di Li, LiHua Wang, ShiPing Song, ChunHai Fan
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  ABSTRACT: A highly sensitive method for the detection of a breast cancer-associated BRCA-1 gene is reported. The detection is based on a classical sandwich-type assay using horseradish peroxidase (HRP) as a catalytic label and electrodeposited Os2+/3+ conducting polymer (PAA-PVI-Os) as a redox mediator. Target DNA could be detected by the HRP-catalyzed reduction of H2O2, leading to a limit of detection as low as 10 fM.
  Science in China Series B Chemistry 04/2012; 52(6):746-750. · 1.20 Impact Factor
• Article: A portable and power-free microfluidic device for rapid and sensitive lead (pb(2+)) detection.

  Chunhui Fan, Shijiang He, Gang Liu, Lianhui Wang, Shiping Song
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  ABSTRACT: A portable and power-free microfluidic device was designed for rapid and sensitive detection of lead (Pb(2+)). 11-mercaptoundecanoic acid (MUA)-functionalized gold nanoparticles (MUA-AuNPs) aggregated in the presence of Pb(2+) for the chelation mechanism. When we performed this analysis on a polydimethylsiloxane (PDMS) microfluidic chip, the aggregations deposited onto the surface of chip and formed dark lines along the laminar flows in the zigzag microchannels. This visual result can be observed by the naked eye through a microscope or just a drop of water as a magnifier. Ten μM Pb(2+) was successfully detected.
  Sensors 01/2012; 12(7):9467-75. · 1.74 Impact Factor
• Article: Using stannous ion as an excellent inorganic ECL coreactant for tris(2,2'-bipyridyl) ruthenium(II).

  Liyan Zheng, Binbin Wang, Yuwu Chi, Shiping Song, Chunhai Fan, Guonan Chen
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  ABSTRACT: It was found that stannous chloride (SnCl(2)), as a popular inorganic reducing reagent, could obviously enhance the electrochemiluminescence (ECL) of tris(2,2'-bipyridyl) ruthenium(II) (Ru(bpy)(3)(2+)) in aqueous solution. Some factors affecting the ECL reactions between Ru(bpy)(3)(2+) and Sn(2+), including pH, concentrations of coreactant, and electrode materials, were investigated by comparison with a classic ECL coreactant tripropylamine (TPA). The Ru(bpy)(3)(2+)-Sn(2+) ECL coreactant system produces stronger and more stable ECL signals, can keep its excellent ECL activity over a wider pH range and has more choices in using electrode materials than the Ru(bpy)(3)(2+)-TPA ECL coreactant system. The ECL mechanism of the Ru(bpy)(3)(2+)-Sn(2+) coreactant system was also studied in detail.
  Dalton Transactions 12/2011; 41(5):1630-4. · 3.84 Impact Factor
• Article: Nanotube-based colorimetric probe for ultrasensitive detection of ataxia telangiectasia mutated protein.

  Qingzhi Zhang, Bin Zhao, Juan Yan, Shiping Song, Rui Min, Chunhai Fan
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  ABSTRACT: We have developed a nanotube-based colorimetric probe using multiwalled carbon nanotubes (MWNTs), anti-immunoglobulin G (anti-IgG), and horseradish peroxidase (HRP). The probe was used as an alternative to conventional colorimetric conjugates to obtain amplified signals in a sandwich-type immunoassay for ataxia telangiectasia mutated (ATM), a potential biomarker for radiation doses and cancers. Results show that the MWNT-based probe colorimetry was 5000 times more sensitive than a conventional ELISA, while its concentration range was 10,000 times wider than that of the latter. Its limit of detection (LOD) was 0.2 fg/mL (54 aM, ~32 molecules in 1 μL samples). Control experiments showed that detection of ATM molecules at the picogram-level could still be achieved in samples that contained protein makers present at more than 100 times the ATM concentration, demonstrating the high specificity of the technique. The MWNT-based probe also has the potential to become a universal probe for colorimetric assays of most protein markers because it can recognize the associated rabbit polyclonal antibodies.
  Analytical Chemistry 12/2011; 83(23):9191-6. · 5.86 Impact Factor
• Article: Universal optical assays based on multi-component nanoprobes for genomic deoxyribonucleic acid and proteins.

  Jiang Li, Ying Wan, Lihua Wang, Xinhua Zhu, Yan Su, Di Li, Yun Zhao, Qing Huang, Shiping Song, Chunhai Fan
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  ABSTRACT: In this report, we developed a universal assay method for both genomic DNA and proteins by using enzyme-based multi-component optical nanoprobes. The nanoprobes are gold nanoparticles assembled with bio-recognizing and signaling elements. We firstly demonstrated that the nanoprobes could detect unpurified asymmetric polymerase chain reaction (PCR) product from genomic DNA of Escherichia coli, with the sensitivity approximately 10 times higher than that of quantitative real-time PCR assay. The limit of detection (LOD) of our nanoprobe-based method is less than 10 pg template DNA (target DNA). Using DNA aptamers as recognition elements, we also showed that as few as 0.1 nM thrombin could be colorimetrically detected with high specificity. These results indicated that the enzyme-based multi-component nanoprobes have the capability to work with real biological samples, and have the potential in various biological and clinical applications.
  Analytica chimica acta 09/2011; 702(1):114-9. · 4.31 Impact Factor
• Article: Carbon nanotube-based ultrasensitive multiplexing electrochemical immunosensor for cancer biomarkers.

  Ying Wan, Wangping Deng, Yan Su, Xinhua Zhu, Cheng Peng, Haiyan Hu, Hongzhen Peng, Shiping Song, Chunhai Fan
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  ABSTRACT: A multiplexing electrochemical immunosensor was developed for ultrasensitive detection of cancer related protein biomarkers. We employed disposable screen-printed carbon electrode (SPCE) array as the detection platform. A universal multi-labeled nanoprobe was developed by loading HRP and goat-anti-rabbit IgG (secondary antibody, Ab(2)) onto multiwalled carbon nanotube (MWNT). This universal nanoprobe was available for virtually any sandwich-based antigen detection and showed superiority in several areas. By using the SPCE array and the universal nanoprobe, we could detect as low as 5 pg mL(-1) of prostate specific antigen (PSA) and 8 pg mL(-1) of Interleukin 8 (IL-8) with the electrochemical immunosensor. We also demonstrated simultaneous detection of two protein biomarkers with this platform. With these attracted features, our immunoassay system shows promising applications for in-field and point-of-care test in clinical diagnostics.
  Biosensors & bioelectronics 09/2011; 30(1):93-9. · 5.43 Impact Factor
• Article: DNA nanostructure-decorated surfaces for enhanced aptamer-target binding and electrochemical cocaine sensors.

  Yanli Wen, Hao Pei, Ying Wan, Yan Su, Qing Huang, Shiping Song, Chunhai Fan
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  ABSTRACT: The sensitivity of aptamer-based electrochemical sensors is often limited by restricted target accessibility and surface-induced perturbation of the aptamer structure, which arise from imperfect packing of probes on the heterogeneous and locally crowded surface. In this study, we have developed an ultrasensitive and highly selective electrochemical aptamer-based cocaine sensor (EACS), based on a DNA nanotechnology-based sensing platform. We have found that the electrode surface decorated with an aptamer probe-pendant tetrahedral DNA nanostructure greatly facilitates cocaine-induced fusion of the split anticocaine aptamer. This novel design leads to a sensitive cocaine sensor with a remarkably low detection limit of 33 nM. It is also important that the tetrahedra-decorated surface is protein-resistant, which not only suits the enzyme-based signal amplification scheme employed in this work, but ensures high selectivity of this sensor when deployed in sera or other adulterated samples.
  Analytical Chemistry 08/2011; 83(19):7418-23. · 5.86 Impact Factor
• Article: A carbon nanotube-based high-sensitivity electrochemical immunosensor for rapid and portable detection of clenbuterol.

  Gang Liu, Haode Chen, Hongzhen Peng, Shiping Song, Jimin Gao, Jianxin Lu, Min Ding, Lanying Li, Shuzhen Ren, Ziying Zou, Chunhai Fan
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  ABSTRACT: Carbon nanotubes have shown their unique advantages of mechanical, chemical and electronic properties in bioanalysis. We herein report a new method to efficiently and reproducibly prepare multi-walled carbon nanotubes (MWNTs)-protein sensing layers for electrochemical immunosensors. This method employs centrifugation to prepare a conjugate of MWNTs and goat anti mouse-immunoglobulin G (IgG) (secondary antibody). The conjugates were then deposited on screen-printed electrodes to form a nanostructured layer (MWNT-I layer). CLB monoclonal antibody was assembled through its binding to the secondary antibody. The MWNT-I layer-based electrodes were used for rapid and sensitive amperometric immunosensing detection of clenbuterol (CLB) in swine urine samples. Horseradish peroxidase-coupled CLB (CLB-HRP) competed with free CLB in the samples to bind the monoclonal antibody. It has shown significantly higher sensitivity and better reproducibility than the chemical conjugation method. This MWNT-based immunosensor is highly sensitive, leading to a limit of detection of 0.1 ng/mL within a rapid assay time of 16 min. Its sensitivity is at least 1 order of magnitude higher than that of a normal immunosensor (without MWNTs). The sensing device is portable with disposable screen-printed electrode, satisfactorily meeting the requirements for field detection of food security-related species.
  Biosensors & bioelectronics 07/2011; 28(1):308-13. · 5.43 Impact Factor
• Source

  Available from: PubMed Central

  Article: Detection of single-nucleotide polymorphism on uidA gene of Escherichia coli by a multiplexed electrochemical DNA biosensor with oligonucleotide-incorporated nonfouling surface.

  Gang Liu, Ruojun Lao, Li Xu, Qin Xu, Lanying Li, Min Zhang, Hao Shen, Sanjay Mathur, Chunhai Fan, Shiping Song
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  ABSTRACT: We report here a practical application of a multiplexed electrochemical DNA sensor for highly specific single-nucleotide polymorphism (SNP) detection. In this work, a 16-electrode array was applied with an oligonucleotide-incorporated nonfouling surfaces (ONS) on each electrode for the resistance of unspecific absorption. The fully matched target DNA templated the ligation between the capture probe assembled on gold electrodes and the tandem signal probe with a biotin moiety, which could be transduced to peroxidase-based catalyzed amperometric signals. A mutant site (T93G) in uidA gene of E. coli was analyzed in PCR amplicons. 10% percentage of single mismatched mutant gene was detected, which clearly proved the selectivity of the multiplexed electrochemical DNA biosensor when practically applied.
  Sensors 01/2011; 11(8):8018-27. · 1.74 Impact Factor