• Title/Summary/Keyword: Loop Detection

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Rapid and Visual Detection of Vibrio parahaemolyticus in Aquatic Foods Using blaCARB-17 Gene-Based Loop-Mediated Isothermal Amplification with Lateral Flow Dipstick (LAMP-LFD)

  • Hu, Yuan-qing;Huang, Xian-hui;Guo, Li-qing;Shen, Zi-chen;LV, Lin-xue;Li, Feng-xia;Zhou, Zan-hu;Zhang, Dan-feng
    • Journal of Microbiology and Biotechnology
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    • v.31 no.12
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    • pp.1672-1683
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    • 2021
  • Vibrio parahaemolyticus is recognized as one of the most important foodborne pathogens responsible for gastroenteritis in humans. The blaCARB-17 gene is an intrinsic β-lactamase gene and a novel species-specific genetic marker of V. parahaemolyticus. In this study, a loop-mediated isothermal amplification (LAMP) assay combined with a lateral flow dipstick (LFD) was developed targeting this blaCARB-17 gene. The specificity of LAMP-LFD was ascertained by detecting V. parahaemolyticus ATCC 17802 and seven other non-V. parahaemolyticus strains. Finally, the practicability of LAMP-LFD was confirmed by detection with V. parahaemolyticus-contaminated samples and natural food samples. The results showed that the optimized reaction parameters of LAMP are as follows: 2.4 mmol/l Mg2+, 0.96 mmol/l dNTPs, 4.8 U Bst DNA polymerase, and an 8:1 ratio of inner primer to outer primer, at 63℃ for 40 min. The optimized reaction time of the LFD assay is 60 min. Cross-reactivity analysis with the seven non-V. parahaemolyticus strains showed that LAMP-LFD was exclusively specific for V. parahaemolyticus. The detection limit of LAMP-LFD for V. parahaemolyticus genomic DNA was 2.1 × 10-4 ng/μl, corresponding to 630 fg/reaction and displaying a sensitivity that is 100-fold higher than that of conventional PCR. LAMP-LFD in a spiking study revealed a detection limit of approximately 6 CFU/ml, which was similar with conventional PCR. The developed LAMP-LFD specifically identified the 10 V. parahaemolyticus isolates from 30 seafood samples, suggesting that this LAMP-LFD may be a suitable diagnostic method for detecting V. parahaemolyticus in aquatic foods.

A Field Deployable Real-Time Loop-Mediated Isothermal Amplification Targeting Five Copy nrdB Gene for the Detection of 'Candidatus Liberibacter asiaticus' in Citrus

  • Tirumalareddy Danda;Jong-Won Park;Kimberly L. Timmons;Mamoudou Setamou;Eliezer S. Louzada;Madhurababu Kunta
    • The Plant Pathology Journal
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    • v.39 no.4
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    • pp.309-318
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    • 2023
  • Huanglongbing (HLB) is one of the most destructive diseases in citrus, which imperils the sustainability of citriculture worldwide. The presumed causal agent of HLB, 'Candidatus Liberibacter asiaticus' (CLas) is a non-culturable phloem-limited α-proteobacterium transmitted by Asian citrus psyllids (ACP, Diaphorina citri Kuwayama). A widely adopted method for HLB diagnosis is based on quantitative real-time polymerase chain reaction (qPCR). Although HLB diagnostic qPCR provides high sensitivity and good reproducibility, it is limited by time-consuming DNA preparation from plant tissue or ACP and the requirement of proper lab instruments including a thermal cycler to conduct qPCR. In an attempt to develop a quick assay that can be deployed in the field for CLas detection, we developed a real-time loop-mediated isothermal amplification (rt-LAMP) assay by targeting the CLas five copy nrdB gene. The rt-LAMP assay using various plant sample types and psyllids successfully detected the nrdB target as low as ~2.6 Log10 copies. Although the rt-LAMP assay was less sensitive than laboratory-based qPCR (detection limit ~10 copies), the data obtained with citrus leaf and bark and ACP showed that the rt-LAMP assay has >96% CLas detection rate, compared to that of laboratory-based qPCR. However, the CLas detection rate in fibrous roots was significantly decreased compared to qPCR due to low CLas titer in some root DNA sample. We also demonstrated that the rt-LAMP assay can be used with a crude leaf DNA extract which is fully deployable in the field for quick and reliable HLB screening.

Dual-Loop Power Control for Single-Phase Grid-Connected Converters with LCL Filter

  • Peng, Shuangjian;Luo, An;Chen, Yandong;Lv, Zhipeng
    • Journal of Power Electronics
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    • v.11 no.4
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    • pp.456-463
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    • 2011
  • Grid-connected converters have widely adopted LCL filters to acquire high harmonic suppression. However, the LCL filter increases the system order so that the design of the system stability would be complicated. Recently, sole-loop control strategies have been used for grid-connected converters with L or LC filters. But if the sole-loop control is directly transplanted to grid-connected converters with LCL filters, the systems may be unstable. This paper presents a novel dual-loop power control strategy composed of a power outer loop and a current inner loop. The outer loop regulates the grid-connected power. The inner loop improves the system stability margin and suppresses the resonant peak caused by the LCL filter. To obtain the control variables, a single-phase current detection is proposed based on PQ theory. The system transfer function is derived in detail and the influence of control gains on the system stability is analyzed with the root locus. Simulation and experimental results demonstrate the feasibility of the proposed control.

Development of a Rapid Assay for Peach Rosette Mosaic Virus Using Loop-mediated Isothermal Amplification (Peach rosette mosaic virus 검출을 위한 신속한 등온증폭법 개발)

  • Lee, Siwon;Lee, Jin-Young;Kim, Jin-Ho;Rho, Jae-Young
    • Microbiology and Biotechnology Letters
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    • v.44 no.4
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    • pp.493-496
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    • 2016
  • Peach rosette mosaic virus (PRMV) is a plant virus that was first reported in 1933 by Peach. It can infect hosts including peach, grape, blueberry, dandelion, plum, cherry tree, and weeds. PRMV is non-reportable in Korea, but it is designated as a controlled virus requiring plant quarantine. In this study, for the rapid and specific detection of PRMV, we developed an assay using loop-mediated isothermal amplification (LAMP). Comparison between conventional polymerase chain reaction (PCR) methods (real time-PCR and nested PCR) and LAMP for the detection of PRMV revealed an equivalent level of sensitivity by all the tested methods. For the LAMP assay, outer primer sets were used to amplify a 264-bp PCR product, which was then digested using the restriction enzyme Pvu II (CAG/CTG), and the visualization of two digestion fragments (207 + 57 bp) indicated a positive reaction. The developed LAMP assay for PRMV is expected to enable the rapid monitoring of PRMV in plants.

VILODE : A Real-Time Visual Loop Closure Detector Using Key Frames and Bag of Words (VILODE : 키 프레임 영상과 시각 단어들을 이용한 실시간 시각 루프 결합 탐지기)

  • Kim, Hyesuk;Kim, Incheol
    • KIPS Transactions on Software and Data Engineering
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    • v.4 no.5
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    • pp.225-230
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    • 2015
  • In this paper, we propose an effective real-time visual loop closure detector, VILODE, which makes use of key frames and bag of visual words (BoW) based on SURF feature points. In order to determine whether the camera has re-visited one of the previously visited places, a loop closure detector has to compare an incoming new image with all previous images collected at every visited place. As the camera passes through new places or locations, the amount of images to be compared continues growing. For this reason, it is difficult for a visual loop closure detector to meet both real-time constraint and high detection accuracy. To address the problem, the proposed system adopts an effective key frame selection strategy which selects and compares only distinct meaningful ones from continuously incoming images during navigation, and so it can reduce greatly image comparisons for loop detection. Moreover, in order to improve detection accuracy and efficiency, the system represents each key frame image as a bag of visual words, and maintains indexes for them using DBoW database system. The experiments with TUM benchmark datasets demonstrates high performance of the proposed visual loop closure detector.

Segmentation-based Signal Processing Algorithm for Vehicle Detection (차량검지를 위한 세그먼트에 기반을 둔 신호처리 알고리즘)

  • Ko, Ki-Won;Woo, Kwang-Joon
    • Proceedings of the KIEE Conference
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    • 2005.10b
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    • pp.306-308
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    • 2005
  • The vehicle detection method using pulse radar has the advantage of maintenance in comparison with loop detection method. We have the information about the vehicle being and position by dividing the signals into sectors in accordance with SSC method, and by applying the discriminant function based on stochastical data. We also reduce the signal processing time.

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Loop-mediated isothermal amplification assay for the rapid detection of swine influenza virus (등온증폭법을 이용한 돼지인플루엔자바이러스 신속 진단법 개발)

  • Kim, Eun-Mi;Jeon, Hyo-Sung;Kim, Ji Jung;Kim, Hee-Jung;Shin, Yeun-Kyung;Song, Jae-Young;Yeo, Sang-Geon;Park, Choi-Kyu
    • Korean Journal of Veterinary Service
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    • v.38 no.2
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    • pp.107-116
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    • 2015
  • In this study, we developed a rapid, sensitive and specific reverse-transcriptase loop-mediated isothermal amplification (RT-LAMP) assay for detection of swine influenza viruse (SIV) including major subtypes of swine influenza viruses H1N1, H1N2 and H3N2, and a novel subtype of influenza A virus that accidentally infected in pig population. The RT-LAMP was completed in 40 min at $58^{\circ}C$ and the sensitivity of the RT-LAMP ($1copy/{\mu}L$) was 10-fold higher than conventional reverse transcription-polymerase chain reaction (RT-PCR) ($10copy/{\mu}L$) and the same to real time RT-PCR ($1copy/{\mu}L$). Also, the result of the RT-LAMP can be confirmed without any detection system. Therefore, the RT-LAMP could be a alternative diagnostic method for SIV detection in national SIV monitoring system and clinical diagnostic laboratory in the future.

Detection of Clostridium difficile by Loop-Mediated Isothermal Amplification (등온증폭법을 이용한 Clostridium difficile 검출)

  • In, Ye-Won;Ha, Su-Jeong;Yang, Seung-Kuk;Oh, Se-Wook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.9
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    • pp.1326-1330
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    • 2012
  • This study was conducted to develop a loop-mediated isothermal amplification (LAMP) method for the detection of Clostridium difficile. The tested target gene was 16S ribosomal RNA. Five different LAMP primer sets were designed, and LAMP was performed. All primer sets targeting the 16S rRNA gene (BIP, FIP, B3, F3, LF, PF) were determined as positive in tcdA-positive, tcdB-postive ($A^+B^+$) and tcdA-negative, tcdB-negative ($A^-B^-$) Clostridium difficile strains. As the LAMP reaction took less than 80 min and did not require expensive machine such as thermocycler, it can be used as a rapid and simple detection method for foodborne pathogens.

Drowsy Driving Detection Algorithm Using a Steering Angle Sensor And State of the Vehicle (조향각센서와 차량상태를 이용한 졸음운전 판단 알고리즘)

  • Moon, Byoung-Joon;Yeon, Kyu-Bong;Lee, Sun-Geol;Hong, Seung-Pyo;Nam, Sang-Yep;Kim, Dong-Han
    • 전자공학회논문지 IE
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    • v.49 no.2
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    • pp.30-39
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    • 2012
  • An effective drowsy driver detection system is needed, because the probability of accident is high for drowsy driving and its severity is high at the time of accident. However, the drowsy driver detection system that uses bio-signals or vision is difficult to be utilized due to high cost. Thus, this paper proposes a drowsy driver detection algorithm by using steering angle sensor, which is attached to the most of vehicles at no additional cost, and vehicle information such as brake switch, throttle position signal, and vehicle speed. The proposed algorithm is based on jerk criterion, which is one of drowsy driver's steering patterns. In this paper, threshold value of each variable is presented and the proposed algorithm is evaluated by using acquired vehicle data from hardware in the loop simulation (HILS) through CAN communication and MATLAB program.

Loop-Mediated Isothermal Amplification Assay Targeting the femA Gene for Rapid Detection of Staphylococcus aureus from Clinical and Food Samples

  • Zhao, Xihong;Li, Yanmei;Park, Myoungsu;Wang, Jun;Zhang, Youhong;He, Xiaowei;Forghani, Fereidoun;Wang, Li;Yu, Guangchao;Oh, Deog-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.23 no.2
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    • pp.246-250
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    • 2013
  • In this study, a loop-mediated isothermal amplification (LAMP) method to rapidly detect Staphylococcus aureus strains was developed and evaluated by extensively applying a large number of S. aureus isolates from clinical and food samples. Six primers were specially designed for recognizing eight distinct sequences on the species-specific femA gene of S. aureus. The detection limits were 100 fg DNA/tube and $10^4$ CFU/ml. The LAMP assay was applied to 432 S. aureus strains isolated from 118 clinical and 314 food samples. Total detection rates for the LAMP and polymerase chain reaction assays were 98.4% (306/311) and 89.4% (278/311), respectively.