• Title/Summary/Keyword: PCR chip

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Design of an Inexpensive Heater using Chip Resistors for a Portable Real-time Microchip PCR System (저항소자를 이용한 휴대형 Real-time PCR 기기용 히터 제작)

  • Choi, Hyoung-jun;Kim, Jeong-tae;Koo, Chi-wan
    • Journal of IKEEE
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    • v.23 no.1
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    • pp.295-301
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    • 2019
  • A heater in a portable real-time polymerase chain reaction(PCR) system is one of the important factors for controlling the PCR thermocycle precisely. Since heaters are integrated on a small-sized PCR chip for rapid heating and fabricated by semiconductor processes, the cost of producing PCR chips is high. Here, we propose to use chip resistors as an inexpensive and accurate temperature control method. The temperature distribution was simulated using one or two chip resistors on a real-time PCR chip and the PCR chip with uniform temperature distribution was fabricated. The temperature rise and fall rates were $18^{\circ}C/s$ and $3^{\circ}C/s$, respectively.

Thermal Design of PCR Chip for LOC (랩온어칩을 위한 중합효소 연쇄반응 칩의 열설계)

  • Kim, Deok-Jong;Kim, Jae-Yun;Park, Sang-Jin;Heo, Pil-U;Yun, Ui-Su
    • 연구논문집
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    • s.33
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    • pp.17-25
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    • 2003
  • In this work, thermal design of a PCR chip for LOC is systematically conducted. From the numerical simulation of a PCR chip based on the finite volume method, how to control the average temperature of a PCR chip and the temperature difference between the denaturation zone and the annealing zone is presented. The average temperature is shown to be controlled by adjusting heat input and a cooler as well as a heater is shown to be necessary to obtain three individual temperature zones for polymerase chain reaction. To reduce the time required, a heat sink for the cooler is not included in the calculation domain for the PCR chip and heat sink design is conducted separately by using a compact modeling method, the porous medium approach.

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Development of Detachable PDMS/Glass PCR-Chip and It's Application to Detection of Male Infertility (분리식 PDMS/유리 중합효소연쇄반응칩 개발 및 유전적 남성불임 검사에의 응용)

  • Ju, Jin-Kyoung;Hwang, Seung-Young;Ahn, Yoo-Min
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.32 no.4
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    • pp.371-377
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    • 2008
  • Our precedent study has reported glass-PDMS (polydimethylsiloxane) based biochip for the gene PCR (polymerase chain reaction). To prevent the contamination of bio sample, the once used biochip must not be used repeatedly. However, the fabrication cost of microheater and microsensor of the biochip was not cheap to use it as a disposable chip. This paper proposes new PCR-chip where the glass substrate integrated with the microheater and microsensor is detachable from the reaction chamber where the sample is injected. That makes it possible to reuse the glass substrate repeatedly. The performance of the proposed detachable PCR-chip was compared with that of the precedent monolithic PCR-chip. The results showed that the SRY (sex determining Y chromosome) gene PCR was successfully performed in the detachable chip compared with the monolithic chip. However, the more efforts to improve the efficiency of surface treatment of PDMS chip are needed to increase the possibility of applying the detachable chip to the detecting of male infertility.

Development of Ultra-Rapid Multiplex PCR Detection against 6 Major Pathogens in Honeybee (꿀벌 6종 주요 병원체에 대한 초고속 다중 PCR 검출법의 개발)

  • Lim, Su-Jin;Kim, Jung-Min;Lee, Chil-Woo;Yoon, Byoung-Su
    • Journal of Apiculture
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    • v.32 no.1
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    • pp.27-39
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    • 2017
  • PCR-chip-based ultra-rapid multiplex PCRs for detection of six major infectious pathogens in honeybee were developed. The 6 kinds of major infectious pathogens in honeybee included Paenibacillus larvae causing American Foulbrood, Melissococcus plutonius causing European Foulbrood as bacteria, Ascosphaera apis (Chalkbrood), Aspergillus flavus (Stonebrood), Nosema apis and Nosema ceranae (Nosemosis) as fungi. The developed PCR-chip-based ultra-rapid multiplex PCR showed successful amplification for all six major pathogens in the presence of more than $10^3$ molecules. The time for confirming amplification (Threshold cycles; Ct-time) was about 7 minutes for two species, and about 9 minutes for four species. Total 40 cycles of PCR took 11 minutes 42 seconds and time for melting point analysis was 1 minute 15 seconds. Total time for whole PCR detection was estimated 12 minutes 57 seconds (40 cycles of PCR and melting point analysis). PCR-chip based ultra-rapid multiplex PCR using standard DNA substrates showed close to 100% accuracy and no false-amplification was found with honeybee genomic DNA. Ultra-rapid multiplex PCR is expected to be a fast and efficient pathogen detection method not only in the laboratory but also in the apiary field.

Characteristics of Indium-Tin-Oxide Electrode for Continuous-flow PCR Chip (연속흐름 중합효소연쇄반응칩 제작을 위한 인듐 산화막 전극의 특성분석)

  • Joung, Seung-Ryong;Kim, Jun-Hyeok;Yi, In-Je;Kang, C.J.;Kim, Yong-Sang
    • The Transactions of The Korean Institute of Electrical Engineers
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    • v.56 no.3
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    • pp.561-565
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    • 2007
  • We propose glass and PDMS (polydimethylsiloxane) chips for DNA amplification with continuous-flow PCR (polymerase chain reaction). The PDMS microchannel was fabricated using a negative molding method for sample injection. Three heaters and sensors of ITO (indium-tin-oxide) thin films were fabricated on glass chip. ITO heaters and sensors were calibrated accurately for the temperature control of the liquid flow. ITO heater generated stable heat versus applied power. ITO sensor resistance was changed linearly versus temperature increase as a RTD (resistance temperature detector) sensor. As a result, we enable precision temperature control of continuous-flow PCR chip. Using the continuous-flow PCR chip DNA plasmid pKS-GFP 720 bp was successfully amplified.

Operation of PCR chip by micropump (마이크로펌프를 이용한 PCR Chip의 구동)

  • 최종필;반준호;장인배;김헌영;김병희
    • Proceedings of the Korean Society of Machine Tool Engineers Conference
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    • 2004.10a
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    • pp.463-467
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    • 2004
  • This paper presents the fabrication possibility of the micro actuator which uses a micro-thermal bubble, generated b micro-heater under pulse heating. The valve-less micropump using the diffuser/nozzle is consists of the lower plate, he middle plate, the upper plate. The lower plate includes the channel and chamber are fabricated on high processability silicon wafer by the DRIE(Deep Reactive Ion Etching) process. The middle plate includes the chamber and diaphragm d the upper plate is the micro-heater. The Micropump is fabricated by bonding process of the three layer. This paper resented the possibility of the PCR chip operation by the fabricated micropump.

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Clinical Usefulness of LabChip Real-time PCR using Lab-On-a-Chip Technology for Diagnosing Malaria

  • Kim, Jeeyong;Lim, Da Hye;Mihn, Do-CiC;Nam, Jeonghun;Jang, Woong Sik;Lim, Chae Seung
    • Parasites, Hosts and Diseases
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    • v.59 no.1
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    • pp.77-82
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    • 2021
  • As malaria remains a major health problem worldwide, various diagnostic tests have been developed, including microscopy-based and rapid diagnostic tests. LabChip real-time PCR (LRP) is a small and portable device used to diagnose malaria using lab-on-a-chip technology. This study aimed to evaluate the diagnostic performance of LRP for detecting malaria parasites. Two hundred thirteen patients and 150 healthy individuals were enrolled from May 2009 to October 2015. A diagnostic detectability of LRP for malaria parasites was compared to that of conventional RT-PCR. Sensitivity of LRP for Plasmodium vivax, P. falciparum, P. malariae, and P. ovale was 95.5%, 96.0%, 100%, and 100%, respectively. Specificity of LRP for P. vivax, P. falciparum, P. malariae, and P. ovale was 100%, 99.3%, 100%, and 100%, respectively. Cohen's Kappa coefficients between LRP and CFX96 for detecting P. vivax, P. falciparum, P. malariae, and P. ovale were 0.96, 0.98, 1.00, and 1.00, respectively. Significant difference was not observed between the results of LRP and conventional RT-PCR and microscopic examination. A time required to amplify DNAs using LRP and conventional RT-PCR was 27 min and 86 min, respectively. LRP amplified DNAs 2 times more fast than conventional RT-PCR due to the faster heat transfer. Therefore, LRP could be employed as a useful tool for detecting malaria parasites in clinical laboratories.

Enteric Virus Detection from Environmental Sample by Oligonucleotide DNA Chip (올리고뉴클레오티드 DNA Chip을 이용한 환경시료에서의 장관계바이러스 검출)

  • 김정미;윤성욱;지영미;윤재득;정용석
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.186-191
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    • 2002
  • The usefulness of oligonucleotide DNA chip was evaluated for detection and primary level identification of major waterborne viruses in environmental samples. The enteric waterborne viruses included enterovirus, adenovirus, and rotavirus. Total intracellular RNA of 10 BGM cell plates showing virus-specific cytopathic effects was extracted at the third day after inoculation. The intracellular RNA was then subjected to either enterovirus-specific RT-PCR followed by sequencing analysis, or the DNA chip. Seven out of 10 positive samples in cell culture were positive but the other three sample were turned out to be negative by both RT-PCR and DNA chip analyses. Nucleotide sequencing results and the DNA chip hybridization results of the RT-PCR product were in complete agreement in the identification of the 7 positive samples as enteroviruses. Using the DNA chip, it took only 3∼4 hr to complete detection and primary level identification of target viruses and additional procedures such as gel electrophoresis or nucleotide sequencing were not necessary. We believe that the DNA chip system can be employed as a highly effective and new detection methodology for environmental viruses.

Rapid Detection for Salmonella spp. by Ultrafast Real-time PCR Assay (Ultrafast Real-time PCR법을 이용한 살모넬라의 신속 검출)

  • Kim, Seok Hwan;Lee, Yu-Si;Joo, In-Sun;Kwak, Hyo Sun;Chung, Gyung Tae;Kim, Soon Han
    • Journal of Food Hygiene and Safety
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    • v.33 no.1
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    • pp.50-57
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    • 2018
  • Salmonella continue to be a major cause of food poisoning worldwide. The rapid detection method of food-borne Salmonella is an important food safety tool. A real-time polymerase chain reaction (PCR) has been used as a rapid method for the detection of pathogens. It has been recently reported that NBS LabChip real-time PCR is a novel, ultrafast, and chip-type-convenient real-time PCR system. We developed the assay method based on NBS LabChip real-time PCR for the rapid detection of Salmonella, which its reaction time was within 20 minutes. Two target genes (invA and stn) were selected to design target specific primers and probes. The new method was validated by checking specificity and sensitivity (limit of detection). This study included forty-two target and twenty-one non-target strains to assess the specificity. This assay was able to identify the 42 Salmonella strains correctly. The limit of detection (LOD) was $10^1copies/{\mu}L$ in Salmonella genomes DNA, while LOD incubated for 4 hr in the inoculated sausage sample ranged from $10^1CFU/g$ to $10^2CFU/g$ as an inoculated cell count. The assay developed in this study could be applied for the investigation of food poisoning pathogens.

PDMS/Glass Serpentine Microchannel Chip for PCR with Bubble Suppression in Sample Injection (시료주입시 기포발생이 억제된 반응조 형태의 중합효소연쇄반응용 PDMS/유리 바이오칩)

  • Cho Chul-Ho;Cho Woong;Hwang Seung-Yong;Ahn Yoo-Min
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.30 no.10 s.253
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    • pp.1261-1268
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    • 2006
  • This paper reports low-cost microreactor $(10{\mu}{\ell})$ biochip for the DNA PCR (polymerase chain reaction). The microbiochip $(20mm{\times}28mm)$ is a hybrid type which is composed of PDMS (polydimethylsiloxane) layer with serpentine micochannel $(360{\mu}m{\times}100{\mu}m)$ chamber and glass substrate integrated with microheater and thermal microsensor. Undesirable bubble is usually created during sample loading to PMDS-based microchip because of hydrophobic chip surface. Created bubbles interrupt stable biochemical reaction. We designed improved microreactor chamber using microfluidic simulation. The designed reactor has a coner-rounded serpentine channel architecture, which enables stable injection into hydrophobic surface using micropipette only. Reactor temperature needed to PCR reaction is controlled within ${\pm}0.5^{\circ}C$ by PID controller of LabVIEW software. It is experimentally confirmed that SRY gene PCR by the fabricated microreactor chip is performed for less than 54 min.