• 제목/요약/키워드: construction sequential analysis

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Integrated Rotary Genetic Analysis Microsystem for Influenza A Virus Detection

  • Jung, Jae Hwan;Park, Byung Hyun;Choi, Seok Jin;Seo, Tae Seok
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2013년도 제45회 하계 정기학술대회 초록집
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    • pp.88-89
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    • 2013
  • A variety of influenza A viruses from animal hosts are continuously prevalent throughout the world which cause human epidemics resulting millions of human infections and enormous industrial and economic damages. Thus, early diagnosis of such pathogen is of paramount importance for biomedical examination and public healthcare screening. To approach this issue, here we propose a fully integrated Rotary genetic analysis system, called Rotary Genetic Analyzer, for on-site detection of influenza A viruses with high speed. The Rotary Genetic Analyzer is made up of four parts including a disposable microchip, a servo motor for precise and high rate spinning of the chip, thermal blocks for temperature control, and a miniaturized optical fluorescence detector as shown Fig. 1. A thermal block made from duralumin is integrated with a film heater at the bottom and a resistance temperature detector (RTD) in the middle. For the efficient performance of RT-PCR, three thermal blocks are placed on the Rotary stage and the temperature of each block is corresponded to the thermal cycling, namely $95^{\circ}C$ (denature), $58^{\circ}C$ (annealing), and $72^{\circ}C$ (extension). Rotary RT-PCR was performed to amplify the target gene which was monitored by an optical fluorescent detector above the extension block. A disposable microdevice (10 cm diameter) consists of a solid-phase extraction based sample pretreatment unit, bead chamber, and 4 ${\mu}L$ of the PCR chamber as shown Fig. 2. The microchip is fabricated using a patterned polycarbonate (PC) sheet with 1 mm thickness and a PC film with 130 ${\mu}m$ thickness, which layers are thermally bonded at $138^{\circ}C$ using acetone vapour. Silicatreated microglass beads with 150~212 ${\mu}L$ diameter are introduced into the sample pretreatment chambers and held in place by weir structure for construction of solid-phase extraction system. Fig. 3 shows strobed images of sequential loading of three samples. Three samples were loaded into the reservoir simultaneously (Fig. 3A), then the influenza A H3N2 viral RNA sample was loaded at 5000 RPM for 10 sec (Fig. 3B). Washing buffer was followed at 5000 RPM for 5 min (Fig. 3C), and angular frequency was decreased to 100 RPM for siphon priming of PCR cocktail to the channel as shown in Figure 3D. Finally the PCR cocktail was loaded to the bead chamber at 2000 RPM for 10 sec, and then RPM was increased up to 5000 RPM for 1 min to obtain the as much as PCR cocktail containing the RNA template (Fig. 3E). In this system, the wastes from RNA samples and washing buffer were transported to the waste chamber, which is fully filled to the chamber with precise optimization. Then, the PCR cocktail was able to transport to the PCR chamber. Fig. 3F shows the final image of the sample pretreatment. PCR cocktail containing RNA template is successfully isolated from waste. To detect the influenza A H3N2 virus, the purified RNA with PCR cocktail in the PCR chamber was amplified by using performed the RNA capture on the proposed microdevice. The fluorescence images were described in Figure 4A at the 0, 40 cycles. The fluorescence signal (40 cycle) was drastically increased confirming the influenza A H3N2 virus. The real-time profiles were successfully obtained using the optical fluorescence detector as shown in Figure 4B. The Rotary PCR and off-chip PCR were compared with same amount of influenza A H3N2 virus. The Ct value of Rotary PCR was smaller than the off-chip PCR without contamination. The whole process of the sample pretreatment and RT-PCR could be accomplished in 30 min on the fully integrated Rotary Genetic Analyzer system. We have demonstrated a fully integrated and portable Rotary Genetic Analyzer for detection of the gene expression of influenza A virus, which has 'Sample-in-answer-out' capability including sample pretreatment, rotary amplification, and optical detection. Target gene amplification was real-time monitored using the integrated Rotary Genetic Analyzer system.

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화성암의 화학적 변질지수에 관한 연구 (A Study on the Chemical Index of Alteration of Igneous Rocks)

  • 최은경;김성욱;김인수;이규환
    • 한국지반공학회논문집
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    • 제28권5호
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    • pp.41-54
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    • 2012
  • 암석이 풍화되어 감에 따라 지반 지지력이 감소되는 것은 잘 알려진 사실이다. 토목 구조물의 시공과 설계에서는 암석의 풍화에 의한 지반정수의 변화가 많이 인용되고 있는데 본 연구는 우리나라 면적의 45.5%에 해당되는 화성암을 대상으로 풍화정도에 따른 화학적 변질지수의 값의 범위와 풍화에 따른 광물조성의 변화를 분석하였다. 대상 시료에 대하여 현재까지 제안된 풍화지수들을 산정하였으며, 유의성 분석을 통해서 지수간의 높은 상관성을 확인하였다. 각각의 풍화지수들은 풍화가 진행될수록 값이 증가 또는 감소하는 경향을 보이는데, 화학적 변질지수(CIA)는 암종이나 광물에 따른 값이 기존연구에서 제시되어 있어 풍화의 정도를 판별하기 용이하므로 본 연구에서는 이를 이용하여 풍화정도를 평가하였다. 풍화에 따른 광물학적 변화는 산성질의 영역에 포함되는 암석은 풍화가 진행되어 감에 따라 기반암-일라이트-녹니석-카올린의 경로를 따르나 고철질의 영역에 포함되는 암석은 풍화 진행에 따라 기반암-스멕타이트-녹니석-카올린의 경로를 주로 따르게 된다. 즉 고철질의 암석에서는 풍화과정에서 양이온교환능이 높은 팽윤성의 점토광물인 스멕타이트의 함량이 증가하는 경향을 보인다. 화학적 풍화지수는 화학성분의 상대적인 변화와 비율을 근거로 작성된 지수이므로 암종별로 값의 범위가 넓게 나타나며, 특정 암종의 대표값을 결정하기 어렵다. 그러나 화학적 풍화지수는 다른 풍화정도를 판별하는 기준들과 마찬가지로 정성적인 기준이 아닌 정량적인 기준으로 풍화도를 평가하는 데 활용이 가능한 것으로 판단된다. 또한 특정지역의 풍화정도를 평가하기 위해서는 화학적 변질지수와 함께 풍화광물의 함량비, 풍화에 따른 강도 특성 등을 함께 고려하는 것이 좋을것으로 판단된다.