• 한국컴퓨터정보학회논문지
• /
• 제18권6호
• /
• pp.1-8
• /
• 2013

본 논문에서는 저가의 소형 PCR 장치에 적합한 펌웨어를 설계하고 구현하였다. 제안된 펌웨어는 실행코드 크기를 최소화하기 위해서 운영체제의 도움을 받지 않고 하드웨어 인터럽트만을 이용하여 실시간 작업들을 동시에 제어한다. 또한 제안된 펌웨어는 usb 통신을 이용하여 PC로부터 동작 과정을 입력받아 마이크로콘트롤러에 연결된 부속장비들을 구동하고, 구동결과를PC로 전달하여 사용자에게 출력하는 주컴퓨터-국소장치 구조에 적합하도록 설계되었다. 제안된 펌웨어를 microchip사의 PIC18F4550 칩에 실제로 탑재하여 저가의 소형 PCR 장치를 제작하였고, 제작한 PCR 장치가 기존 상용 PCR 장치는 제작 비용과 부피를 대폭 줄이면서도 유사한 DNA 증폭 결과를 보임을 확인하였다.

• 한국진공학회:학술대회논문집
• /
• 한국진공학회 2013년도 제45회 하계 정기학술대회 초록집
• /
• pp.273-273
• /
• 2013

Recently, Point-of-care (POC) testing microdevices enable to do the patient monitoring, drug screening, pathogen detection in the outside of hospital. Immunochromatographic strip (ICS) is one of the diagnostic technologies which are widely applied to POC detection. Relatively low cost, simplicity to use, easy interpretations of the diagnostic results and high stability under any circumstances are representative advantages of POC diagnosis. It would provide colorimetric results more conveniently, if the genetic analysis microsystem incorporates the ICS as a detector part. In this work, we develop a reverse transcriptase-polymerase chain reaction (RT-PCR) microfluidic device integrated with a ROSGENE strip for colorimetric influenza H1N1 virus detection. The integrated RT-PCR- ROSGENE device is consist of four functional units which are a pneumatic micropump for sample loading, 2 ${\mu}L$ volume RT-PCR chamber for target gene amplification, a resistance temperature detector (RTD) electrode for temperature control, and a ROSGENE strip for target gene detection. The device was fabricated by combining four layers: First wafer is for RTD microfabrication, the second wafer is for PCR chamber at the bottom and micropump channel on the top, the third is the monolithic PDMS, and the fourth is the manifold for micropump operation. The RT-PCR was performed with subtype specific forward and reverse primers which were labeled with Texas-red, serving as a fluorescent hapten. A biotin-dUTP was used to insert biotin moieties in the PCR amplicons, during the RT-PCR. The RT-PCR amplicons were loaded in the sample application area, and they were conjugated with Au NP-labeled hapten-antibody. The test band embedded with streptavidins captures the biotin labeled amplicons and we can see violet colorimetric signals if the target gene was amplified with the control line. The off-chip RT-PCR amplicons of the influenza H1N1 virus were analyzed with a ROSGENE strip in comparison with an agarose gel electrophoresis. The intensities of test line was proportional to the template quantity and the detection sensitivity of the strip was better than that of the agarose gel. The test band of the ROSGENE strip could be observed with only 10 copies of a RNA template by the naked eyes. For the on-chip RT-PCR-ROSGENE experiments, a RT-PCR cocktail was injected into the chamber from the inlet reservoir to the waste outlet by the micro-pump actuation. After filling without bubbles inside the chamber, a RT-PCR thermal cycling was executed for 2 hours with all the microvalves closed to isolate the PCR chamber. After thermal cycling, the RT-PCR product was delivered to the attached ROSGENE strip through the outlet reservoir. After dropping 40 ${\mu}L$ of an eluant buffer at the end of the strip, the violet test line was detected as a H1N1 virus indicator, while the negative experiment only revealed a control line and while the positive experiment a control and a test line was appeared.

• 제어로봇시스템학회논문지
• /
• 제12권5호
• /
• pp.419-424
• /
• 2006

In this study, a thermal control system which applied a Peltier device for the polymerase chain reaction(PCR) machine is to be designed. Here in order for it to easily follow the characteristics of the thermal cycle existing for gene amplification of the PCR sample, a PCR control board utilizing a thermal sensor, a Peltier, and a 8 bit microprocessor is made up. Especially a fuzzy type PD control algorithm is applied periodically in time response, and control system is implemented. For that matter, the characteristic data of subject system is obtained and analysed to begin with. Based on this analysed data, the proposed control algorithm is applied and an evaluation of the performance of the whole system take place through the PC.

• 대한의용생체공학회:의공학회지
• /
• 제44권6호
• /
• pp.465-475
• /
• 2023

In this study, we demonstrated a silica nanofibrous membrane based on the electrospinning process and evaluated its DNA isolation and purification performance in PCR pretreatment. Generally, silica membranes made of non-woven fabric are used for PCR pretreatment, but this study aimed to improve the efficiency of the pretreatment process by developing a nanofiber-type silica membrane with high specific surface area and porosity. In order to manufacture a nanofiber-shaped silica film while maintaining the original physical properties of silica, nanofiber membranes produced by adding various concentrations of PEO (5 wt%, 8 wt%, and 10 wt%) to silica prepared by the sol-gel method were compared. In terms of nanofiber membrane production, the higher the PEO concentration, the more effective it was in producing nanofiber membranes. The produced silica nanofiber membrane was inserted to a pretreatment device used in commercial PCR equipment, and the pretreatment performance was compared and verified using Salmonella bacteria. When Salmonella was used, samples containing 5 wt% PEO showed superior PCR efficiency compared to samples containing 8 wt% and 10 wt% PEO. These results show that adding 5 wt% of PEO can effectively improve DNA purification and separation by producing a nanofiber-shaped silica film while maintaining the physical properties of silica. We expect that this study will contribute to the development of effective PCR pretreatment technology essential for various molecular biology applications.

• Parasites, Hosts and Diseases
• /
• 제59권1호
• /
• pp.77-82
• /
• 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.

• The Plant Pathology Journal
• /
• 제40권1호
• /
• pp.40-47
• /
• 2024

Garlic can be infected by a variety of viruses, but mixed infections with leek yellow stripe virus, onion yellow dwarf virus, and allexiviruses are the most damaging, so an easy, inexpensive on-site method to simultaneously detect at least these three viruses with a certain degree of accuracy is needed to produce virus-free plants. The most common laboratory method for diagnosis is multiplex reverse transcription polymerase chain reaction (RT-PCR). However, allexiviruses are highly diverse even within the same species, making it difficult to design universal PCR primers for all garlic-growing regions in the world. To solve this problem, we developed an inexpensive on-site detection system for the three garlic viruses that uses a commercial mobile PCR device and a compact electrophoresis system with a blue light. In this system, virus-specific bands generated by electrophoresis can be identified by eye in real time because the PCR products are labeled with a fluorescent dye, FITC. Because the electrophoresis step might eventually be replaced with a lateral flow assay (LFA), we also demonstrated that a uniplex LFA can be used for virus detection; however, multiplexing and a significant cost reduction are needed before it can be used for on-site detection.

• 대한기계학회논문집A
• /
• 제34권12호
• /
• pp.1785-1791
• /
• 2010

중합효소 연쇄반응(PCR)을 수행하려면 세포 용해(cell lysis)와 DNA추출(DNA purification)과정이 포함된 시료 전처리 과정을 거쳐야 한다. 종래의 시료 전처리 과정은 계면활성제와 같은 세포용해 버퍼를 이용하거나 열 또는 전기적 방법으로 세포막 파열을 유도하여 세포벽을 깬 후에 잔여물 처리과정을 거쳐 DNA를 추출하게 된다. 본 연구에서는 마이크로 비드와 PDMS 기둥을 이용한 필터가 있는 시료 전처리용 바이오칩을 설계 및 제작하였다. 또한 제작된 바이오칩을 사용하여 $80^{\circ}C$에서 2분간 세포용해를 수행하고 DNA를 추출하였다. 칩에서 전처리과정을 거친 시료내의 DNA농도와 순도를 측정하고 DNA PCR과 겔 전기영동을 통해 시료 전처리용 바이오칩의 성능을 평가하였다.

• 한국재료학회지
• /
• 제34권2호
• /
• pp.63-71
• /
• 2024

Slipchip offers advantages such as high-throughout, low cost, and simple operation, and therefore, it is one of the technologies with the greatest potential for high-throughput, single-cell, and single-molecule analyses. Slipchip devices have achieved remarkable advances over the past decades, with its simplified molecular diagnostics gaining particular attention, especially during the COVID-19 pandemic and in various infectious diseases scenarios. Medical testing based on nucleic acid amplification in the Slipchip has become a promising alternative simple and rapid diagnostic tool in field situations. Herein, we present a comprehensive review of Slipchip device advances in molecular diagnostics, highlighting its use in digital recombinase polymerase amplification (RPA), loop-mediated isothermal amplification (LAMP), and polymerase chain reaction (PCR). Slipchip technology allows users to conduct reliable droplet transfers with high-throughput potential for single-cell and molecule analyses. This review explores the device's versatility in miniaturized and rapid molecular diagnostics. A complete Slipchip device can be operated without special equipment or skilled handling, and provides high-throughput results in minimum settings. This review focuses on recent developments and Slipchip device challenges that need to be addressed for further advancements in microfluidics technology.

• 청정기술
• /
• 제9권3호
• /
• pp.101-105
• /
• 2003

MEMS(Micro Electro Mechanical System) 기술에서 실리콘 식각기술의 중요성으로 플라즈마 식각기술의 개발이 꾸준히 진행되고 있다. 본 연구에서는 ICP(Inductive Coupled Plasma)를 이용하여 플라즈마를 발생시켜, 이온에너지를 증가시키지 않고도 이온밀도를 높이고 이온입자들에 의한 식각의 방향성을 가할 수 있는 새로운 플라즈마 기술을 이용하였다. 이같이 플라즈마를 이용하여 실리콘웨이퍼를 식각하여 제조하는 MEMS 응용분야는 다양하나, 본 연구에서는 미생물배양에 응용할 수 있는 PCR(Polymerase Chain Reaction)장치 제작을 위한 식각에 이용하였다. Platen power, Coil power 및 Process pressure에 다양한 변화를 주어 각 변수에 따른 식각속도를 관찰하였다. 각 공정별 변수를 변화시킨 결과 Platen 12W, Coil power 500W, 식각/Passivation Cycle 6/7sec 일 경우 식각속도는 $1.2{\mu}m/min$ 이었고, sidewall profile은 $90{\pm}0.7^{\circ}$로 나타나 매우 우수한 결과를 보였다. 분 연구에 SF6를 식각에 이용하였으며 공정의 최적화를 통하여 사용량을 최소화하여환경영향이 최소가 될 수 있는 가능성이 있었다.

• 농업생명과학연구
• /
• 제43권6호
• /
• pp.105-109
• /
• 2009

경산 묘목단지는 대단위 과수종묘를 생산하고 있어 경쟁력 강화를 위하여 무독묘 보증 생산이 요구되고 있다. 특히 생산규모가 큰 사과종묘에 대한 빠르고 정확한 바이러스 진단이 시급하다. 본 연구에서는 사과바이러스 진단을 위하여 다량의 시료를 동시 분쇄할 수 있는 bead beater를 이용하였으며 분쇄 bead는 저가의 산업용 glass bead (0.4 mm 직경)를 일회용으로 채택하였다. RNA추출을 위하여서는 guanidine thiocyanate 용액이 Trizol 용액보다 효과적인 것으로 나타났으며 silica membrane tube의 이용으로 RNA추출 간편성을 높일 수 있었다. 사과바이러스는 RT-PCR에 의하여 검증하였다.