• Title/Summary/Keyword: electrochemical DNA chip

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Detection of Human Papillomavirus in Male and Female Urine by Electrochemical DNA Chip and PCR Sequencing

  • Nilyanimit, Pornjarim;Wanlapakorn, Nasamon;Niruthisard, Somchai;Pohthipornthawat, Natkrita;Karalak, Anant;Laowahutanont, Piyawat;Phanuphak, Nittaya;Gemma, Nobuhiro;Poovorawan, Yong
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.9
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    • pp.5519-5525
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    • 2013
  • Background: Cervical cancer is the second most common cancer in Thai women after breast cancer. Currently, the Papanicolaou (Pap) smear is the recommended procedure for cervical cancer screening in Thailand, but only a relatively small percentage of women follow this screening program. An alternative method to detect HPV genotypes associated with cervical cancer is self-sampling of urine, which is a more widely accepted method. Our study aimed to evaluate the prevalence of HPV in Thai women using urine and cervical swabs and prevalence of HPV in Thai men using urine samples. Materials and Methods: Tumorigenic HPV detection was accomplished by electrochemical DNA chip and PCR/direct sequencing. In addition to HPV prevalence, we report the concordance between different methods and sample types. One-hundred and sixteen women and 100 men were recruited. Histological examination revealed normal cytology in 52 women, atypical squamous cells of undetermined significance (ASCUS) in 9, low-grade squamous intraepithelial lesions (LSIL) in 24, and high-grade squamous intraepithelial lesions (HSIL) in 31. One-hundred men were classified as heterosexuals (n=45) and homosexuals (n=55). Results: The most prevalent HPV genotype in our study was HPV16. The HPV detection rate was generally lower in urine samples compared with cervical samples. Overall, there was good agreement for the detection of carcinogenic HPV from female cervical samples between the DNA chip and PCR/sequencing, with 88.8% total agreement and a kappa value of 0.76. In male urine samples, the level of agreement was higher in heterosexuals compared with homosexuals. Conclusions: Further improvement is required to increase an overall yield of HPV DNA detection in urine samples before clinical application of a urine-based HPV screening program. The electrochemical DNA chip test is a promising technique for carcinogenic HPV detection.

Genome Detection Using an Integrated type DNA Chip Microelectrode-array and Non-labeling Target DNA (집적형 DNA칩 미소 전극 어레이 및 비수식화 표적 DNA를 이용한 유전자 검출)

  • Choi, Yong-Sung;Lee, Hea-Yeon;Tanaka, Hiroyuki;Tanaka, Hidekafu;Kwon, Young-Soo;Kawai, Tomoii
    • Proceedings of the KIEE Conference
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    • 2001.11a
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    • pp.274-276
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    • 2001
  • This research aims to develop the multiple channel electrochemical DNA chip using microfabrication technology. At first, we fabricated a high integration type DNA chip array by lithography technology. Several probe DNAs consisting of thiol group at their 5-end were immobilized on the sold electrodes. Then target DNAs were hybridized and reacted. Cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. Therefore, it is able to detect a plural genes electrochemically after immobilization of a plural probe DNA and hybridization of non-labeling target DNA on the electrodes simultaneously. It suggested that this DNA chip could recognize the sequence specific genes.

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Electrochemical Gene Detection Using Hoechat Groove Binder (Hoechst groove binder를 이용한 유전자의 전기화학적 검출)

  • Choi, Yong-Sung;Lee, Woo-Ki;Lee, Kyung-Sup
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2006.05a
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    • pp.65-70
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    • 2006
  • In this study, a DNA chip with a microelectrode array was fabricated using microfabrication technology. Several probe DNAs consisting of mercaptohexyl moiety at their 5 end were immobilized on the gold electrodes by DNA arrayer. Then target DNAs were hybridized and reacted with Hoechst 33258, which is a DNA minor groove binder and electrochemically active dye. Linear sweep voltammetry or cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. It was derived from Hoechst 33258 concentrated at the electrode surface through association with formed hybrid. It suggested that this DNA chip could recognize the sequence specific genes.

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Detection of Sequence-Specific Gene by Multi-Channel Electrochemical DNA Chips

  • Zhang, Xuzhi;Ji, Xinming;Cui, Zhengguo;Yang, Bing;Huang, Jie
    • Bulletin of the Korean Chemical Society
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    • v.33 no.1
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    • pp.69-75
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    • 2012
  • Five-channel electrochemical chips were fabricated based on the Micro-electromechanical System (MEMS) technology and were used as platforms to develop DNA arrays. Different kinds of thiolated DNA strands, whose sequences were related to white spot syndrome virus (WSSV) gene, were separately immobilized onto different working electrodes to fabricate a combinatorial biosensor system. As a result, different kinds of target DNA could be analyzed on one chip via a simultaneous recognition process using potassium ferricyanide as an indicator. To perform quantitative target DNA detection, a limit of 70 nM (S/N=3) was found in the presence of 600 nM coexisting noncomplementary ssDNA. The real samples of loop-mediated isothermal amplification (LAMP) products were detected by the proposed method with satisfactory result, suggesting that the multichannel chips had the potential for a high effective microdevice to recognize specific gene sequence for pointof-care applications.

A Study on the Development of Electrochemical Biochip (전기화학적 바이오칩의 개발에 관한 연구)

  • Choi, Yong-Sung;Kwon, Young-Soo;Park, Dae-Hee
    • Proceedings of the KIEE Conference
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    • 2003.10a
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    • pp.300-302
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    • 2003
  • This research aims to develop the multi-channel type label-free DNA chip that has the above characteristics and be able to solve the problems. At first, we fabricated a high integrated type DNA chip array by lithography technology. It is able to detect a various genes electrochemically after immobilization of a various probe DNA and hybridization of label-free target DNA on the electrode s simultaneously.

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A Study on Electrical Properties of Dendrimer (미소전극형 DNA칩 어레이를 이용한 유전자의 검출)

  • Choi, Yong-Sung;Lee, Kyung-Sup
    • Proceedings of the KIEE Conference
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    • 2006.07c
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    • pp.1324-1326
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    • 2006
  • In this study, an integrated microelectrode array was fabricated on glass slide using microfabrication technology. Probe DNAs consisting of mercaptohexyl moiety at their 5-end were spotted on the gold electrode using micropipette or DNA arrayer utilizing the affinity between gold and sulfur. Cyclic voltammetry in 5mM ferricyanide/ferrocyanide solution at 100 mV/s confirmed the immobilization of probe DNA on the gold electrodes. When several DNAs were detected electrochemically, there was a difference between target DNA and control DNA in the anodic peak current values. It was derived from specific binding of Hoechst 33258 to the double stranded DNA due to hybridization of target DNA. It suggested that this DNA chip could recognize the sequence specific genes. It suggested that multichannel electrochemical DNA microarray is useful to develop a portable device for clinical gene diagnostic system.

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Voltammetric Analysis on a Disposable Microfluidic Electrochemical Cell

  • Chand, Rohit;Han, Dawoon;Kim, Yong-Sang
    • Bulletin of the Korean Chemical Society
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    • v.34 no.4
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    • pp.1175-1180
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    • 2013
  • A microfabricated electrochemical cell comprising PDMS-based microchannel and in-channel gold microelectrodes was fabricated as a sensitive and a miniature alternative to the conventional electroanalytical systems. A reproducible fabrication procedure enabled patterning of multiple microelectrodes integrated within a PDMS-based fluidic network. The active area of each electrode was $200{\mu}m{\times}200{\mu}m$ with a gap of $200{\mu}m$ between the electrodes which resulted in a higher signal to noise ratio. Also, the PDMS layer served the purpose of shielding the electrical interferences to the measurements. Analytes such as potassium ferrocyanide; amino acid: cysteine and nucleoside: guanosine were characterized using the fabricated cell. The microchip was comparable to bulk electrochemical systems and its applicability was also demonstrated with flow injection based rapid amperometric detection of DNA samples. The device so developed shall find use as a disposable electrochemical cell for rapid and sensitive analysis of electroactive species in various industrial and research applications.