• Title/Summary/Keyword: internal amplification control

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Detection of HCV-RNA by Reverse Transcription Polymerase Chain Reaction Using Biotinylated and Radioiodinated Primers (역전사 중합효소 연쇄반응(RT-PCR)에 의한 HCV-RNA의 검출 : Biotin 및 방사성옥소 표지 Primer로 구성된 Kit의 이용)

  • Ryu, Jin-Sook;Moon, Dae-Hyuk;Cheon, Jun-Hong;Chung, Yoon-Young;Park, Hung-Dong;Chung, Young-Hwa;Lee, Young-Sang
    • The Korean Journal of Nuclear Medicine
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    • v.28 no.2
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    • pp.220-226
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    • 1994
  • This study was performed to evaluate the clinical applicability of the reverse transcription polymerase chain reaction (RT-PCR ) kit of HCV-RNA using biotinylated and radioiodinated primers. Study subjects were 118 patients with positive anti-HCV. HCV-RNA in patient's serum was extracted by guanidium thiocyanate method. After first amplification, the product was reamplified by primers labelled with biotin and I-125. The final amplification product was defected by counting the radioactivity after incubation in avidin coated tubes. In 51 samples, the test was repeated for evaluation of reproducibility. This new method was also compared with conventional RT-PCR methods in 34 samples from patients with chronic liver disease. The results were as follows ; 1) HCV-RNA was positive in 85(97%)of 88 patients with chronic liver disease, and in 23 (73%) of 30 patients with normal liver function. 2) In comparison with conventional method, HCV-RNA was detected in 32(94%) of 34 patients with new method, whereas in 27(79%) of the same group with conventional method. 3) Repeated test with new method in 52 samples demonstrated 82% of concordant result. In conclusion, new method with biotinylated and radioiodinated primers was more sensitive than conventional method. However, great care must be taken for quality control because there were considerable interassay variation and possiblity of false positivity and false negativity.

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Design of a Compact GPS/MEMS IMU Integrated Navigation Receiver Module for High Dynamic Environment (고기동 환경에 적용 가능한 소형 GPS/MEMS IMU 통합항법 수신모듈 설계)

  • Jeong, Koo-yong;Park, Dae-young;Kim, Seong-min;Lee, Jong-hyuk
    • Journal of Advanced Navigation Technology
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    • v.25 no.1
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    • pp.68-77
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    • 2021
  • In this paper, a GPS/MEMS IMU integrated navigation receiver module capable of operating in a high dynamic environment is designed and fabricated, and the results is confirmed. The designed module is composed of RF receiver unit, inertial measurement unit, signal processing unit, correlator, and navigation S/W. The RF receiver performs the functions of low noise amplification, frequency conversion, filtering, and automatic gain control. The inertial measurement unit collects measurement data from a MEMS class IMU applied with a 3-axis gyroscope, accelerometer, and geomagnetic sensor. In addition, it provides an interface to transmit to the navigation S/W. The signal processing unit and the correlator is implemented with FPGA logic to perform filtering and corrrelation value calculation. Navigation S/W is implemented using the internal CPU of the FPGA. The size of the manufactured module is 95.0×85.0×.12.5mm, the weight is 110g, and the navigation accuracy performance within the specification is confirmed in an environment of 1200m/s and acceleration of 10g.

The Effect of Vitamin E on Bleomycin-Induced Pulmonary Injury and Fibrosis in Rat - Comparison of Penicillamine- or Deferoxamine-Treated Group - (백서에서 Bleomycin 투여로 인한 폐손상 및 폐섬유화에 대한 Vitamin E의 영향 - Penicillamine, Deferoxamine 투여군과 비교 -)

  • Jung, Soon-Hee;Yong, Suk-Joong;Ahn, Chul-Min;Shin, Kye-Chul;Choi, In-Joon;Cho, Sang-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.2
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    • pp.184-205
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    • 1995
  • Background: Pulmonary toxicity by bleomycin has multiple mechanisms including direct tissue toxicity due to oxygen-derived free radicals and indirect toxicity through amplification of pulmonary inflammation. To evaluate the effect of chelators or free radical scavenger to lung damage induced by bleomycin, penicillamine as a copper chelator, deferoxamine as an iron chelator and vitamin E as a free radical scavenger were administered. Methods: Two hundred Wistar rats were divided into five groups: Control, bleomycin treated, bleomycin-penicillamine treated, bleomycin-deferoxamine treated, and bleomycin-vitamin E treated groups. Rats sacrificed on day 1, day 3, day 4, day 7, day 14, and day 28 after treatment. Bronchoalveolar lavage, light microscopic and immunohistologic studies for type I, III, IV collagens, fibronectin, laminin and NBD phallicidin were evaluated. Results: There was a significant increase in the total cell counts of bronchoalveolar lavage on day 1 from all treated animals and vitamin treated group showed an abrupt decrease in total cell counts with decrease of neutrophils on day 3. Bleomycin-vitamin E treated group had the least histologic changes such as pulmonary fibrosis. The alveolar basement membranes were positive for type IV collegen and laminin. Basement membranes of bleomycin, bleomycin-penicillamine, or bleomycin-deferoxamine treated groups were disrupted and fragmented on day 4 or 7. The bleomycin-vitamin E treated group had intact basement membranes until day 28. Conclusion: Bleomycin-induced pulmonary fibrosis was related to the severity of acute injury to oxygen radicals or activation of neutrophils and disruption of basement membrane. Vitamin E seemed to be the most effective antioxidant in the inhibition of bleomycin-induced pulmonary injury and fibrosis.

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Deterimination of an Optimal Time Point for Analyzing Transcriptional Activity and Analysis of Transcripts of Avian Influenza Virus H9N2 in Cultured Cell (배양세포에서 Semi-quantitative RT-PCR에 의한 조류인플루엔자 H9N2의 전사활성 분석 최적 시기 결정 및 전사체 분석)

  • Na, Gi-Youn;Lee, Young-Min;Byun, Sung-June;Jeon, Ik-Soo;Park, Jong-Hyeon;Cho, In-Soo;Joo, Yi-Seok;Lee, Yun-Jung;Kwon, Jun-Hun;Koo, Yong-Bum
    • Korean Journal of Microbiology
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    • v.45 no.3
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    • pp.286-290
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    • 2009
  • The transcription of mRNA of avian influenza virus is regulated temporally during infection. Therefore, the measurement of transcript level in host cells should be performed before viral release from host cells because errors can occur in the analysis of the transcript levels if the viruses released from the infected cells re-infect cells. In this study, the timing of viral release was determined by measuring the level of viral RNA from viruses released from H9N2-infected chicken fibroblast cell line UMNSAH/DF-1 by semi-quantitative RT-PCR. The viral genomic RNA was isolated together with mouse total RNA which was added to the collected medium as carrier to monitor the viral RNA recovery and to use its GAPDH as an internal control for normalizing reverse transcription reaction as well as PCR reaction. It was found that viral release of H9N2 in the chicken fibroblast cell line UMNSAH/DF-1 took between 16 and 20 h after infection. We measured all 8 viral mRNA levels. Of the 8 transcripts, 7 species of viral mRNAs (each encoding HA, NA, PB1, PB2, NP, M, NS, respectively) except PA mRNA showed robust amplification, indicating these mRNA can be used as targets for amplification to measure transcript levels. These results altogether suggest that the method in this study can be used for screening antiviral materials against viral RNA polymerase as a therapeutic target.