• Title/Summary/Keyword: Lateral Flow

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Development of a Lateral Flow Strip-Based Recombinase Polymerase Amplification Assay for the Detection of Haemonchus contortus in Goat Feces

  • Wu, Yao-Dong;Wang, Qi-Qi;Wang, Meng;Elsheikha, Hany M.;Yang, Xin;Hu, Min;Zhu, Xing-Quan;Xu, Min-Jun
    • Parasites, Hosts and Diseases
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    • v.59 no.2
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    • pp.167-171
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    • 2021
  • Haemonchosis remains a significant problem in small ruminants. In this study, the assay of recombinase polymerase amplification (RPA) combined with the lateral flow strip (LFS-RPA) was established for the rapid detection of Haemonchus contortus in goat feces. The assay used primers and a probe targeting a specific sequence in the ITS-2 gene. We compared the performance of the LFS-RPA assay to a PCR assay. The LFS-RPA had a detection limit of 10 fg DNA, which was 10 times less compared to the lowest detection limit obtained by PCR. Out of 24 goat fecal samples, LFS-RPA assay detected H. contortus DNA with 95.8% sensitivity, compared to PCR, 79.1% sensitivity. LFS-RPA assay did not detect DNA from other related helminth species and demonstrated an adequate tolerance to inhibitors present in the goat feces. Taken together, our results suggest that LFS-RPA assay had a high diagnostic accuracy for the rapid detection of H. contortus and merits further evaluation.

Recent Research Trend in Lateral Flow Immunoassay Strip (LFIA) with Colorimetric Method for Detection of Cancer Biomarkers (암 바이오마커 검출용 비색법 기반 측면 흐름 면역 크로마토그래피 분석법(LFIA) 스트립의 최신 연구 동향)

  • Lee, Sooyoung;Lee, Hye Jin
    • Applied Chemistry for Engineering
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    • v.31 no.6
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    • pp.585-590
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    • 2020
  • Successful early diagnosis of cancer diseases such as lung, prostate, liver and adrenocortical carcinoma is a key step in determining the cost of treatment, survival rate, and cure rate. Most of current cancer diagnosis systems including biopsy, computed tomography (CT), positron emission tomography (PET)-CT, magnetic resonance imaging (MRI), ultrasonography, etc., require expensive and complicated equipment with highly trained human resources. Global medical and scientific communities have thus made numerous efforts on developing effective but rapid disease management system via introducing a wide spectrum of point-of-care medical diagnosis devices. Among them, a lateral flow immunoassay strip technique has gained a great attention due to many advantages such as cost-effectiveness, short inspection time, and user friendly accessibility. In this mini-review, we will highlight recent research trend on the development of colorimetry based LFIA strips for cancer diagnosis and discuss the future research direction and potential applications.

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.

Application of Rapid and Reliable Detection of Cymbidium Mosaic Virus by Reverse Transcription Recombinase Polymerase Amplification Combined with Lateral Flow Immunoassay

  • Do-Hyun, Kim;Rae-Dong, Jeong;Sena, Choi;Ho-Jong, Ju;Ju-Yeon, Yoon
    • The Plant Pathology Journal
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    • v.38 no.6
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    • pp.665-672
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    • 2022
  • Cymbidium mosaic virus (CymMV) is one of economically important viruses that cause significant losses of orchids in the world. In the present study, a reverse transcription recombinase polymerase amplification (RT-RPA) assay combined with a lateral flow immunostrip (LFI) assay was developed for the detection of CymMV in orchid plants. A pair of primers containing fluorescent probes at each terminus that amplifies highly specifically a part of the coat protein gene of CymMV was determined for RT-RPA assay. The RT-RPA assay involved incubation at an isothermal temperature (39℃) and could be performed rapidly within 30 min. In addition, no cross-reactivity was observed to occur with odontoglossum ringspot virus and cymbidium chlorotic mosaic virus. The RT-RPA with LFI assay (RT-RPA-LFI) for CymMV showed 100 times more sensitivity than conventional reverse transcription polymerase chain reaction (RT-PCR). Furthermore, the RT-PCR-LFI assay demonstrated the simplicity and the rapidity of CymMV detection since the assay did not require any equipment, by comparing results with those of conventional RT-PCR. On-site application of the RT-RPA-LFI assay was validated for the detection of CymMV in field-collected orchids, indicating a simple, rapid, sensitive, and reliable method for detecting CymMV in orchids.

Development of Recombinase Polymerase Amplification Combined with Lateral Flow Strips for Rapid Detection of Cowpea Mild Mottle Virus

  • Xinyang Wu;Shuting Chen;Zixin Zhang;Yihan Zhang;Pingmei Li;Xinyi Chen;Miaomiao Liu;Qian Lu;Zhongyi Li;Zhongyan Wei;Pei Xu
    • The Plant Pathology Journal
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    • v.39 no.5
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    • pp.486-493
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    • 2023
  • Cowpea mild mottle virus (CPMMV) is a global plant virus that poses a threat to the production and quality of legume crops. Early and accurate diagnosis is essential for effective managing CPMMV outbreaks. With the advancement in isothermal recombinase polymerase amplification and lateral flow strips technologies, more rapid and sensitive methods have become available for detecting this pathogen. In this study, we have developed a reverse transcription recombinase polymerase amplification combined with lateral flow strips (RT-RPA-LFS) method for the detection of CPMMV, specifically targeting the CPMMV coat protein (CP) gene. The RT-RPA-LFS assay only requires 20 min at 40℃ and demonstrates high specificity. Its detection limit was 10 copies/µl, which is approximately up to 100 times more sensitive than RT-PCR on agarose gel electrophoresis. The developed RT-RPA-LFS method offers a rapid, convenient, and sensitive approach for field detection of CPMMV, which contribute to controlling the spread of the virus.

An Experimental Study on the Estimation of Optimum Length of Soil Flow Protector with Wall Stiffness (벽체 강성에 따른 토사유입차단판의 최적 길이 산정에 관한 실험적 연구)

  • Yoo, Jae-Won;Seo, Min-Su;Son, Su-Won;Im, Jong-Chul
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.39 no.6
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    • pp.789-799
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    • 2019
  • The settlement hardly occurs in structures supported by pile foundation such as abutment, culvert but a cavity is formed in the lower part of a structure. As a result, soil discharged from the lateral ground to the cavity accelerates the settlement of the lateral ground of the structure, resulting in a larger settlement. Therefore, in order to prevent problems caused by cavity under the structure supported by pile foundation, soil Flow Protector (briefly called 'FLP'), which can be easily installed on the side of structure, was developed. In this study, an laboratory model test was carried out to prove the reduction effect of settlement and to estimate the optimal installation length of the FLP. As a result, the installation of the FLP reduced the settlement of the lateral ground and prevented the leakage of lateral ground soil into the cavity. If the stiffness of the FLP is small, the state or active earth pressure is generated in the upper part, which is not favorable for stability. But if the stiffness of the FLP is high enough, the passive earth pressure area is generated in the upper part, which will be advantageous for the stability. Also, the increased installation length of FLP is effective to reduce the settlement. And the ratio of the optimal length of the FLP to the box structure height (H = 250 mm) are flexible FLP 1.38, stiff FLP 0.73.

Prediction of Stage Discharge Curve and Lateral Distribution of Unit Discharge in an Arbitrary Cross Section Channel with Floodplain Vegetation (홍수터 식생을 고려한 불규칙한 단면에서의 수위-유량 곡선 및 단위유량 횡분포 예측)

  • Kim, Tae-Beom;Jang, Ji-Yeon;Shin, Jae-Kook;Choi, Sung-Uk
    • Journal of Korea Water Resources Association
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    • v.44 no.2
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    • pp.157-167
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    • 2011
  • A numerical model was developed to predict the stage-discharge curve and lateral distribution of unit discharge in open channels with nonuniform cross section or compound open-channels. The governing equation is the one-dimensional momentum equation based on assumptions of the steady and uniform flow conditions in the longitudinal direction and the uniform water surface elevation in a cross section. Vegetative drag force term was included in governing equation in order to reflect the effect of floodplain vegetation on the flow characteristics. Finite element method was applied to obtain the numerical solution of the governing equation. Stage-discharge curve and lateral distribution of unit discharge for a given water surface are calculated based on input data, such as the cross sectional geometry, Manning's roughness coefficient, vegetative information and longitudinal slope of channel bed. The developed model was verified by comparing the calculated results with the observed data and the results of Darby and Thorne's(1996) model and the nonlinear k-$\epsilon$ model. The verified model was applied to estimate the upstream boundary conditions in two-dimensional flow model. The numerical results using laterally distributed unit discharge were compared with those obtained using uniformly distributed unit discharge in two-dimensional flow model.

Detection of Genetically Modified Soybean in Tofu and Biji using PCR and Immunological Methods (PCR 방법과 면역학적 분석법을 이용한 두부와 비지에서 GM 콩의 검출법)

  • Kim, Myo-Young;Kim, Jae-Hwan;Kim, Hae-Yeong
    • Applied Biological Chemistry
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    • v.48 no.1
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    • pp.77-81
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    • 2005
  • To monitor GM soybean in soybean processed foods, tofu and biji, we prepared tofu and biji containing 0%, 1%, 3%, 5% and 100% GM soybean, respectively. We examined epsps gene inserted in soybean by PCR and EPSPS protein expressed in soybean using western blotting and lateral flow strip test to compare the sensitivity of these methods. A PCR product of 123 bp inserted in GM soybean was detected in all tofu and biji containing 1%, 3%, 5% and 100% GM soybean with the exception of 0% samples; however, the size of 600 bp inserted in GM soybean was only detected in tofu containing 100% soybean and in biji containing 5% and 100% soybean. In the protein level, GM soybean product was only detected in tofu and biji containing 100% GM soybean by western blotting. In addition, only biji containing 100% GM soybean was detected by lateral flow strip test. We concluded that in order to detect GM soybean efficiently in processed food, the PCR method is more sensitive than immunological methods. With the PCR method, small size product with approximately 100 bp in PCR product is sensitive to detect GM soybean in processed foods.

Centrifuge Modeling on Lateral Flow of Soft Soils and Displacement of Bridge Abutment on the Composite Ground (복합지반상 교대변위 및 지반 측방유동에 관한 원심모델링)

  • Heo, Yol;Park, Sunghun;Yun, Seokhyun;Kwon, Seonuk
    • Journal of the Korean GEO-environmental Society
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    • v.8 no.5
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    • pp.39-46
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    • 2007
  • In this study, the centrifuge tests were performed to investigate the lateral flow behavior and stability of the ground improved by SCP. The centrifuge tests were fulfilled in the case of the back of abutment filled by EPS (case 1) and soil (case 2), and the potentiometer was installed on the abutment and embankment to measure the vertical and horizontal displacement at the top of abutment. As a result, the vertical displacement measured at the back of abutment was maximum 2.1 m, which was about 12% if compared with the height of embankment. In the case of the back of abutment filled by soil, the vertical and horizontal displacement measured at the top of abutment was 10 cm and 1.1 m, respectively, which exceeded the allowable horizontal displacement. On the other hand, in the case of the back of abutment filled by EPS, the vertical displacement of abutment did nor occur and the horizontal displacement was 1.4 cm. Therefore, the effect of SCP improvement with EPS method adopted to prevent the lateral flow and assure the stability of embankment on the soft ground was far superior.

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Analysis of Flood Characteristics at Confluence by Lateral Inflow (횡유입에 의한 합류부 홍수특성 분석)

  • Choi, Hung-Sik;Cho, Min-Suk;Park, Young-Seop
    • Journal of the Korean Society of Hazard Mitigation
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    • v.6 no.1 s.20
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    • pp.59-68
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    • 2006
  • Flow separation of recirculation zone by increasing of flow and change of its direction at confluence results in backwater due to conveyance reduction. The hydraulic characteristics of flow separation are analysed by experimental results of flow ratios of tributary and main streams and approaching angles. The boundary of flow separation by dimensionless length and width is defined by the streamline of zero and this definition agrees well to the existing investigation. Because flow separation doesn't appear in small flow ratio and approaching angle of $30^{\circ}$, the equation of flow separation with flow ratio and approaching angle is provided. In flow separation consideration and comparing with previous results, the existing equations of dimensionless length and width ratios by function of approaching angle, flow ratio, and downstream Froude number are modified and also contraction coefficient and shape factor are analysed. Dimensionless length and width ratios are proportional to the flow ratio and approaching angle. In analysis of water surface profiles, the backwater effects are proportional to the flow ratio and approaching angle and the magnitude at outside wall is greater than that of inside wall of main stream. The length, $X_l$ from the beginning of confluence to downstream of uniform flow, where the depth is equal to uniform depth, is characterized by width of stream, flow ratio, approaching angle, and contraction coefficient. The ratios between maximum water depth by backwater and minimum depth at separation are analysed.