• Title/Summary/Keyword: Limit of Detection

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Application of Engineered Zinc Finger Proteins Immobilized on Paramagnetic Beads for Multiplexed Detection of Pathogenic DNA

  • Shim, Jiyoung;Williams, Langley;Kim, Dohyun;Ko, Kisung;Kim, Moon-Soo
    • Journal of Microbiology and Biotechnology
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    • v.31 no.9
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    • pp.1323-1329
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    • 2021
  • Micro-scale magnetic beads are widely used for isolation of proteins, DNA, and cells, leading to the development of in vitro diagnostics. Efficient isolation of target biomolecules is one of the keys to developing a simple and rapid point-of-care diagnostic. A zinc finger protein (ZFP) is a double-stranded (ds) DNA-binding domain, providing a useful scaffold for direct reading of the sequence information. Here, we utilized two engineered ZFPs (Stx2-268 and SEB-435) to detect the Shiga toxin (stx2) gene and the staphylococcal enterotoxin B (seb) gene present in foodborne pathogens, Escherichia coli O157 and Staphylococcus aureus, respectively. Engineered ZFPs are immobilized on a paramagnetic bead as a detection platform to efficiently isolate the target dsDNA-ZFP bound complex. The small paramagnetic beads provide a high surface area to volume ratio, allowing more ZFPs to be immobilized on the beads, which leads to increased target DNA detection. The fluorescence signal was measured upon ZFP binding to fluorophore-labeled target dsDNA. In this study, our system provided a detection limit of ≤ 60 fmol and demonstrated high specificity with multiplexing capability, suggesting a potential for development into a simple and reliable diagnostic for detecting multiple pathogens without target amplification.

Defect Detection in Laser Welding Using Multidimensional Discretization and Event-Codification (Multidimensional Discretization과 Event-Codification 기법을 이용한 레이저 용접 불량 검출)

  • Baek, Su Jeong;Oh, Rocku;Kim, Duck Young
    • Journal of the Korean Society for Precision Engineering
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    • v.32 no.11
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    • pp.989-995
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    • 2015
  • In the literature, various stochastic anomaly detection methods, such as limit checking and PCA-based approaches, have been applied to weld defect detection. However, it is still a challenge to identify meaningful defect patterns from very limited sensor signals of laser welding, characterized by intermittent, discontinuous, very short, and non-stationary random signals. In order to effectively analyze the physical characteristics of laser weld signals: plasma intensity, weld pool temperature, and back reflection, we first transform the raw data of laser weld signals into the form of event logs. This is done by multidimensional discretization and event-codification, after which the event logs are decoded to extract weld defect patterns by $Na{\ddot{i}}ve$ Bayes classifier. The performance of the proposed method is examined in comparison with the commercial solution of PRECITEC's LWM$^{TM}$ and the most recent PCA-based detection method. The results show higher performance of the proposed method in terms of sensitivity (1.00) and specificity (0.98).

Graphene Based Electrochemical DNA Biosensor for Detection of False Smut of Rice (Ustilaginoidea virens)

  • Rana, Kritika;Mittal, Jagjiwan;Narang, Jagriti;Mishra, Annu;Pudake, Ramesh Namdeo
    • The Plant Pathology Journal
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    • v.37 no.3
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    • pp.291-298
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    • 2021
  • False smut caused by Ustilaginoidea virens is an important rice fungal disease that significantly decreases its production. In the recent past, conventional methods have been developed for its detection that is time-consuming and need high-cost equipments. The research and development in nanotechnology have made it possible to assemble efficient recognition interfaces in biosensors. In this study, we present a simple, sensitive, and selective oxidized graphene-based geno-biosensor for the detection of rice false smut. The biosensor has been developed using a probe DNA as a biological recognition element on paper electrodes, and oxidized graphene to enhance the limit of detection and sensitivity of the sensor. Probe single-stranded DNA (ssDNA) and target ssDNA hybridization on the interface surface has been quantitatively measured with the electrochemical analysis tools namely, cyclic voltammetry, and linear sweep voltammetry. To confirm the selectivity of the device, probe hybridization with non-complementary ssDNA target has been studied. In our study, the developed sensor was able to detect up to 10 fM of target ssDNA. The paper electrodes were employed to produce an effective and cost-effective platform for the immobilization of the DNA and can be extended to design low-cost biosensors for the detection of the other plant pathogens.

Development of Leak Detection System of Heat Exchanger using Acoustic Emission Technique (음향방출기법을 이용한 열교환기 누설검출시스템 개발)

  • Lee, Min-Rae;Lee, Jun-Hyeon
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.26 no.5
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    • pp.864-871
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    • 2002
  • Acoustic emission(AE) technique has been applied to not only mechanical property testing but also on-line monitoring of the entire structure or a limit zone only. Although several AE devices have already been developed for on-line monitoring, the price of these systems is very high and it is difficult for the field to apply yet. In this study, we developed a specially designed PC-based leak detection system using A/D board. In this paper, AE technique has been applied to detect leak for heat exchanger by analyzing the characteristics of signal obtained from leak. It was confirmed that the characteristics of the signal generated by the turbulence of gas in the heat exchanger is narrow band signal having between 130-250kHz. Generally, the amplitude of leak signal is increased as the leak size increasing, but showed no significant change at frequency characteristic. Leak source location can be found by determining for the paint of highest signal amplitude by comparing with several fixed sensors. In this paper, AE results are compared with the PC-based leak detection system using A/D board.

A Novel Active Anti-islanding Method for Grid-connected Photovoltaic Inverter

  • Jung, Young-Seok;Choi, Jae-Ho;Yu, Gwon-Jong
    • Journal of Power Electronics
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    • v.7 no.1
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    • pp.64-71
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    • 2007
  • This paper proposes a novel active frequency drift (AFD) method to improve the islanding detection performance with minimum current harmonics. To detect the islanding phenomenon of grid-connected photovoltaic (PV) inverters concerning the safety hazards and possible damage to other electric equipment, anti-islanding methods have been described. The AFD method that uses chopping fraction (cf) enables the islanding detection to drift up (or down) the frequency of the voltage during the islanding situation. However, the performance of the conventional AFD method is inefficient and causes difficulty in designing the appropriate cf value to meet the limit of harmonics. In this paper, the periodic chopping fraction based on a novel AFD method is proposed. This proposed method shows the analytical design value of cf to meet the test procedure of IEEE Std. 929-2000 with power quality and islanding detection time. To verify the validation of the proposed method, the islanding test results are presented. It is confirmed that the proposed method has not only less harmonic distortion but also better performance of islanding detection compared with the conventional AFD method.

Detection of Aspergillus, Penicillium, and Fusarium Species by Sandwich Enzyme-Linked Immunosorbent Assay Using Mixed Monoclonal Antibodies

  • Kwak, Bo-Yeon;Kwon, Byung-Joon;Kweon, Chang-Hee;Shon, Dong-Hwa
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.385-389
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    • 2004
  • The antibody-mix sandwich enzyme-linked immunosorbent assay (Ab-mix sELlS A) system was developed in order to simultaneously detect the extracellular polysaccharide (FPS) of Aspergillus, Penicillium, or Fusarium species using one detection system. The detection limit and detection range of the Ab-mix sELISA towards EPS of Penicilliun citrinum were not changed, and those towards Fusarium moniliforme EPS were changed a little compared to that of individual sandwich ELISA [9, 10]. The fungal culture filtrates of Aspergillus and Penicillium species showed nearly similar reactivity towards Ab-mix sELISA as that of sELISA using the MAb lB8 alone [9]. Also, the fungal culture filtrates of Fusarium species showed nearly the same reactivity towards Ab-mix sELISA as that of sELISA using the MAb lB8 alone [10]. Thus, this ELISA system showed that the three genera of molds, Aspergillus, Penicillium, or Fusarium, which are three major important molds producing mycotoxins in food or agricultural commodities, could be detected at the same time, using one detection system.

Detection of Norovirus in Contaminated Ham by Reverse Transcriptase-PCR and Nested PCR

  • Kim, Seok-Ryel;Kim, Du-Woon;Kwon, Ki-Sung;Hwang, In-Gyun;Oh, Myung-Joo
    • Food Science and Biotechnology
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    • v.17 no.3
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    • pp.651-654
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    • 2008
  • In order to enhance the efficacy of norovirus detection by reverse transcriptase-polymerase chain reaction (RT-PCR) and nested PCR, this study developed a norovirus mRNA concentration method using poly oligo dT-conjugated magnetic beads. An efficient norovirus detection protocol was performed on commercial ham using 2 viral elution buffers (glycine buffer and Tris beef extract buffer) and 2 concentration solutions [polyethylene glycol (PEG) and zirconium hydroxide]. The different approaches were verified by RT-PCR and nested PCR. This method was performed on ham in less than 8 hr by artificial inoculation of serial dilutions of the virus ranging from 1,000 to 1 RT-PCR unit/mL. The viral extraction and concentration method had 10-fold higher sensitivity using the combination of Tris beef extract buffer and PEG as compared to glycine buffer and zirconium hydroxide. This method proved that RT-PCR and nested PCR have the sensitive ability to detect norovirus in commercial ham, in that norovirus was successfully detected in artificially contaminated samples at a detection level as low as 1-10 RT-PCR unit/mL. Overall, such a detection limit suggests this protocol is both quick and efficient in terms of its potential use for detecting norovirus in meat products.

Anodic Stripping Voltammetric Detection of Arsenic(III) at Platinum-Iron(III) Nanoparticle Modified Carbon Nanotube on Glassy Carbon Electrode

  • Shin, Seung-Hyun;Hong, Hun-Gi
    • Bulletin of the Korean Chemical Society
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    • v.31 no.11
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    • pp.3077-3083
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    • 2010
  • The electrochemical detection of As(III) was investigated on a platinum-iron(III) nanoparticles modified multiwalled carbon nanotube on glassy carbon electrode(nanoPt-Fe(III)/MWCNT/GCE) in 0.1 M $H_2SO_4$. The nanoPt-Fe(III)/MWCNT/GCE was prepared via continuous potential cycling in the range from -0.8 to 0.7 V (vs. Ag/AgCl), in 0.1 M KCl solution containing 0.9 mM $K_2PtCl_6$ and 0.6 mM $FeCl_3$. The Pt nanoparticles and iron oxide were co-electrodeposited into the MWCNT-Nafion composite film on GCE. The resulting electrode was examined by cyclic voltammetry (CV), scanning electron microscopy (SEM), and anodic stripping voltammetry (ASV). For the detection of As(III), the nanoPt-Fe(III)/MWCNT/GCE showed low detection limit of 10 nM (0.75 ppb) and high sensitivity of $4.76\;{\mu}A{\mu}M^{-1}$, while the World Health Organization's guideline value of arsenic for drinking water is 10 ppb. It is worth to note that the electrode presents no interference from copper ion, which is the most serious interfering species in arsenic detection.

Rapid Detection of Salmonella spp. in Fresh-Cut Cabbage by Real-Time PCR (Real-Time PCR을 이용한 신선편이 양배추에서 Salmonella spp.의 신속검출)

  • Bang, Mi-Kyung;Park, Seung-Ju;Kim, Yun-Ji;Kim, Ji-Gang;Oh, Se-Wook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.10
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    • pp.1522-1527
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    • 2010
  • This study was conducted to find out the minimal time needed for detection of Salmonella spp. which exist at very low concentration in foods by using real-time PCR. The sal-F and sal-R sequences were used as primers and sal-P was used as a probe. The detection limit of Salmonella spp. was $3.77{\times}10^2\;cfu/mL$ in buffered peptone water (BPW). Microbial growth was monitored after artificially inoculated Salmonella spp. into BPW. The obtained growth curve was well fitted with the equation, y=$0.0127x^2$+0.5927x-0.4317 ($R^2$=0.99), if assuming that 1 cell exists in 25 g sample (0.04 cfu/mL). The microbial concentration will be reduced to 10 fold by adding BPW during sample treatment, so actual initial concentration at the starting point of enrichment is 0.004 cfu/mL. At this condition, real-time PCR detection would be possible only when microbial concentration increase occurs to exceed the detection limit (377 cfu/mL). The time needed for microbial increase was calculated from the growth curve equation as 7 hours and 20 minutes. Therefore the total time required for detection was less than 10 hours including the PCR operating time.

Effect of Pre-treatments on the Content of Heavy Metals in Packaging Paper

  • Jo, Byoung-Muk;Jeong, Myung-Joon
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 2006.06b
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    • pp.465-469
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    • 2006
  • Pre-treatment methods to determine various heavy metal contents in packaging papers were investigated by ICP-ES (Inductively Coupled Plasma Emission Spectrometry) analysis. Pre-treatment methods utilized in this study include dry ashing and decomposition methods ($HNO_{3-}HClO_{4-}HF,\;HNO_{3},\;and\;H_{2}SO_{4-}HNO_{3}$). They were compared with the conventional extraction (water) and migration (3% acetic acid) methods. The five representative heavy metals (Cd, As, Pb, Cr and Hg) were analyzed. For Cd, Hg, and As, the results were below detection limit of the instrument. In case of Cr and Pb, the migration test is considered to be a better method compared to the extraction test, but all pretreated methods showed much higher detection efficiency than the extraction or migration test. However, the detection ratio between the migration test and decomposition methods was different. Among all decomposition methods, the nitric acid - perchloric acid - hydrofluoric acid treatment brought a slightly higher detection value than others, but there was no significant difference among them except sulfuric acid - nitric acid method. Concerning Pb, the sulfuric acid - nitric acid method showed a low detection efficiency compared to other decomposition methods. The sulfuric acid - nitric acid method is, thus, not considered to be a suitable analysis method for Pb in packaging papers.

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