• Title/Summary/Keyword: Molecular Detection

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Ultra-Sensitive Analysis of Microcystin LR Using Microchip Based Detection System

  • Pyo, Dong-Jin;Huang, Yan;Kim, Young-Min;Hahn, Jong-Hoon
    • Bulletin of the Korean Chemical Society
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    • v.26 no.6
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    • pp.939-942
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    • 2005
  • For the detection of cyanobacterial toxin, an Enzyme-linked immunosorbent assay (ELISA) was integrated into a PDMS microchip. The conjugates of microcystin-LR (MCLR) and keyhole limpet hemocyanin (KLH) were adsorbed on the surface of polystyrene beads and these MCLR-KLH polystyrene beads were introduced into a microchamber. MCLR on the surface of polystyrene beads reacted with horseradish peroxides (HRP) conjugated anti-MCLR monoclonal antibody (mAb) which had a competitive reaction with MCLR in water sample. After the enzyme substrate 3,3,5,5-tetramethyl benzidine (TMB) was injected into the chamber and catalyzed by HRP, the color change was detected with a liquid-cord waveguide. This integration shortened the conventional ELISA analysis time from several hours to about 30 min with only 4.2 $\mu$L MCLR sample consuming which was useful for the environmental analysis. More over, troublesome operations required for ELISA could be replaced by simple operations. The microchip based detection system showed a good sensitivity of 0.05 $\mu$g/L and maintained good reliability through its quantitative range with low coefficients of variation (2.5-10.5%).

Doppler LIDAR Measurement of Wind in the Stratosphere

  • Dong, Jihui;Cha, Hyun-Ki;Kim, Duk-Hyeon;Baik, Sung-Hoon;Wang, Guocheng;Tang, Lei;Shu, Zhifeng;Xu, Wenjing;Hu, Dongdong;Sun, Dongsong
    • Journal of the Optical Society of Korea
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    • v.14 no.3
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    • pp.199-203
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    • 2010
  • A mobile direct detection Doppler LIDAR based on molecular backscattering for measurement of wind in the stratosphere has been developed in Hefei, China. First, the principle of wind measurement with direct detection Doppler LIDAR is presented. Then the configuration of the LIDAR system is described. Finally, the primary experimental results are provided and analyzed. The results indicate that the detection range of the designed Doppler LIDAR reached 50 km altitude, and there is good consistency between the molecular Doppler wind LIDAR(DWL) and the wind profile radar(WPR) in the low troposphere.

Development of a Quantitative Real-time Nucleic Acid Sequence based Amplification (NASBA) Assay for Early Detection of Apple scar skin viroid

  • Heo, Seong;Kim, Hyun Ran;Lee, Hee Jae
    • The Plant Pathology Journal
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    • v.35 no.2
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    • pp.164-171
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    • 2019
  • An assay for detecting Apple scar skin viroid (ASSVd) was developed based on nucleic acid sequence based amplification (NASBA) in combination with realtime detection during the amplification process using molecular beacon. The ASSVd specific primers for amplification of the viroid RNA and molecular beacon for detecting the viroid were designed based on highly conserved regions of several ASSVd sequences including Korean isolate. The assay had a detection range of $1{\times}10^4$ to $1{\times}10^{12}$ ASSVd RNA $copies/{\mu}l$ with reproducibility and precision. Following the construction of standard curves based on time to positive (TTP) value for the serial dilutions ranging from $1{\times}10^7$ to $1{\times}10^{12}$ copies of the recombinant plasmid, a standard regression line was constructed by plotting the TTP values versus the logarithm of the starting ASSVd RNA copy number of 10-fold dilutions each. Compared to the established RT-PCR methods, our method was more sensitive for detecting ASSVd. The real-time quantitative NASBA method will be fast, sensitive, and reliable for routine diagnosis and selection of viroid-free stock materials. Furthermore, real-time quantitative NASBA may be especially useful for detecting low levels in apple trees with early viroid-infection stage and for monitoring the influence on tree growth.

Possibility of Using DNA Chip Technology for Diagnosis of Human Papillomavirus

  • Liu, Cui-Hua;Ma, Wen-Li;Shi, Rong;Ou, Yang-Qian;Zhang, Bao;Zheng, Wen-Ling
    • BMB Reports
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    • v.36 no.4
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    • pp.349-353
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    • 2003
  • To explore the application of DNA chip technology for the detection and typing of Human Papillomavirus (HPV), the HPV6, 11, 16 and 18 gene fragments were isolated and printed onto aminosilane-coated glass slides by a PixSys 5500 microarrayer as probes to prepare the HPV gene chips. HPV samples, after being labeled with fluorescent dye by restriction display PCR (RD-PCR) technology, were hybridized with the microarray, which was followed by scanning and analysis. The experimental condition for preparing the HPV gene chips was investigated, and the possibility of HPV genotyping using gene chips was discussed. The technique that was established in this study for preparing HPV gene chips is practical. The results of the present study demonstrated the versatility and inspiring prospect of using this technology to detect and genotype HPV.

Array-based Nano-amplification Technique Was Applied in Detection of Hepatitis E Virus

  • Liu, Hui-Hui;Cao, Xuan;Yang, Yong;Liu, Ming-Gui;Wang, Ye-Fu
    • BMB Reports
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    • v.39 no.3
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    • pp.247-252
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    • 2006
  • A rapid method for the detection of Hepatitis E Virus (HEV) was developed by utilizing nano-gold labeled oligonucleotide probes, silver stain enhancement and the microarray technique. The 5'-end -$NH_2$ modified oligonucleotide probes were immobilized on the surface of the chip base as the capture probe. The detection probe was made of the 3'-end -SH modified oligonucleotide probe and nano-gold colloid. The optimal concentrations of these two probes were determined. To test the detection sensitivity and specificity of this technique, a conservative fragment of the virus RNA was amplified by the RT-PCR/PCR one step amplification. The cDNA was hybridized with the capture probes and the detection probes on microarray. The detection signal was amplified by silver stain enhancement and could be identified by naked eyes. 100 fM of amplicon could be detected out on the microarray. As the results, preparation of nano-gold was improved and faster. Development time also was shortened to 2 min. Thus, considering high efficiency, low cost, good specificity and high sensitivity, this technique is alternative for the detection of HEV.

Mycobacterium tuberculosis DNA Detection and Molecular Drug Susceptibility Test in AFB-stained Sputum Slides

  • Jung, Dongju;Lee, Hyeyoung;Park, Sangjung
    • Biomedical Science Letters
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    • v.22 no.1
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    • pp.24-28
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    • 2016
  • Tuberculosis (TB) remains an unsolved community health problem since identification of its causing microorganism called Mycobacterium tuberculosis (MTB) by Robert Koch in 1882. Annually, eight million TB cases are newly reported and 2~3 million patients die from TB. Pulmonary TB is highly infectious and untreated pulmonary TB patients are believed to infect >10 people in a year. The conventional methods for diagnosis of TB are chest X-ray and isolation of the causing microorganisms from patient specimens. Screening of TB is conducted with smeared sputum in slides, and TB is confirmed by identification of MTB in cultured specimens. One of the fatal pitfalls of screening detection for smeared sputum is that it is impossible to distinguish MTB and other acid-fast bacilli (AFB) because they are stained equally with Ziehl-Neelsen (ZN) stain. Culture of MTB is the most reliable method for diagnosis of TB but it takes 4~8 weeks. In this report, we suggest a fast and highly-reliable MTB detection method that distinguishes AFB in sputum samples. Purified DNA from the AFB stained slide samples offered by The Korean Institute of Tuberculosis were used to detect infected MTB in patients. PCR, real-time PCR and reverse blot hybridization assay (REBA) methods were applied to purified DNA. Conclusively, the real-time PCR method was confirmed to produce high sensitivity and we were able to further detect drug-resistant MTB with REBA.

The Cause Analysis of Pitting Corrosion on the Waterjet Impeller (물분사 추진기 임펠러 부식에 대한 원인분석)

  • Lee, Hyeong-Sin;Jung, Un-Hwa
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.6
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    • pp.545-551
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    • 2020
  • Cause analysis of surface pitting crack on a waterjet impeller was conducted. The waterjet impeller was made from stainless steel duplex 2205, which is more resistant to corrosion and local corrosion than typical stainless steel 316L and 317L, and has high mechanical strength, making it a useful material in various marine structures and seawater desalination facilities. The measurements were taken by scanning electron microscopy (SEM) and molecular ecological detection. The chemical composition of S was examined by SEM in the area of pitting corrosion. The dsrAB gene was detected on the sample of the pitting corrosion of the impeller through molecular ecological detection. Therefore, pitting corrosion on the surface of a waterjet impeller was caused by sulphite-reducing bacteria (SRB). To prevent the spread of SRB, management is required through high temperature treatments (over 65℃), pH management, or the insulation of a hull and waterjet.