• Title/Summary/Keyword: Sensitive detection

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Development of an Indirect Non-Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Cronobacter muytjensii in Infant Formula Powder (유아용 조제분유 내 Cronobacter muytjensii 검지를 위한 간접 비경합 면역분석법의 개발)

  • Song, Xinjie;Kim, Myunghee
    • The Korean Journal of Food And Nutrition
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    • v.26 no.4
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    • pp.936-944
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    • 2013
  • Cronobacter muytjensii is an important foodborne pathogen as a potential risk in infant formula powder (IFP). To develop a new and sensitive method for the detection of Cronobacter spp. in IFP, an immunoglobulin G (IgG) specific for C. muytjensii (formerly known as Enterobacter sakazakii ATCC 51329) was developed. Further, an indirect noncompetitive enzyme-linked immunosorbent assay (INC-ELISA) was developed by using the anti-C. muytjensii IgG. As a result, this newly developed INC-ELISA method was found very sensitive for C. muytjensii with detection limit of $6.5{\times}10^3CFU/ml$ in pure culture and 1 cell/25 g of IFP. This INC-ELISA method also displayed excellent specificity for C. muytjensii showing no cross-reactivity with other strains of Cronobacter genus and 11 other foodborne pathogenic strains. These results show that the developed INC-ELISA method was very sensitive, efficient, and rapid for the detection of C. muytjensii. Hence, this method could be applied to the development of diagnostic kits for the rapid and easy detection of C. muytjensii.

Loop-mediated Isothermal Amplification assay for Detection of Candidatus Liberibacter Asiaticus, a Causal Agent of Citrus Huanglongbing

  • Choi, Cheol Woo;Hyun, Jae Wook;Hwang, Rok Yeon;Powell, Charles A
    • The Plant Pathology Journal
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    • v.34 no.6
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    • pp.499-505
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    • 2018
  • Huanglongbing (HLB, Citrus greening disease) is one of the most devastating diseases that threaten citrus production worldwide. Although HLB presents systemically, low titer and uneven distribution of these bacteria within infected plants can make reliable detection difficult. It was known loop-mediated isothermal amplification (LAMP) method has the advantages of being highly specific, rapid, efficient, and laborsaving for detection of plant pathogens. We developed a new LAMP method targeting gene contained tandem repeat for more rapid and sensitive detection of Candidatus Liberibacter asiaticus (CLas), putative causal agent of the citrus huanglongbing. This new LAMP method was 10 folds more sensitive than conventional PCR in detecting the HLB pathogen and similar to that of real-time PCR in visual detection assay by adding SYBR Green I to mixture and 1% agarose gel electrophoresis. Positive reactions were achieved in reaction temperature 57, 60 and $62^{\circ}C$ but not $65^{\circ}C$. Although this LAMP method was not more sensitive than real-time PCR, it does not require a thermocycler for amplification or agarose gel electrophoresis for resolution. Thus, we expect that this LAMP method shows strong promise as a reliable, rapid, and cost-effective method of detecting the CLas in citrus and can be applied for rapid diagnosis is needed.

Damage detection of mono-coupled multistory buildings: Numerical and experimental investigations

  • Xu, Y.L.;Zhu, Hongping;Chen, J.
    • Structural Engineering and Mechanics
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    • v.18 no.6
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    • pp.709-729
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    • 2004
  • This paper presents numerical and experimental investigations on damage detection of mono-coupled multistory buildings using natural frequency as only diagnostic parameter. Frequency equation of a mono-coupled multistory building is first derived using the transfer matrix method. Closed-form sensitivity equation is established to relate the relative change in the stiffness of each story to the relative changes in the natural frequencies of the building. Damage detection is then performed using the sensitivity equation with its special features and minimizing the norm of an objective function with an inequality constraint. Numerical and experimental investigations are finally conducted on a mono-coupled 3-story building model as an application of the proposed algorithm, in which the influence of modeling error on the degree of accuracy of damage detection is discussed. A mono-coupled 10-story building is further used to examine the capability of the proposed algorithm against measurement noise and incomplete measured natural frequencies. The results obtained demonstrate that changes in story stiffness can be satisfactorily detected, located, and quantified if all sensitive natural frequencies to damaged stories are available. The proposed damage detection algorithm is not sensitive to measurement noise and modeling error.

Real-time structural damage detection using wireless sensing and monitoring system

  • Lu, Kung-Chun;Loh, Chin-Hsiung;Yang, Yuan-Sen;Lynch, Jerome P.;Law, K.H.
    • Smart Structures and Systems
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    • v.4 no.6
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    • pp.759-777
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    • 2008
  • A wireless sensing system is designed for application to structural monitoring and damage detection applications. Embedded in the wireless monitoring module is a two-tier prediction model, the auto-regressive (AR) and the autoregressive model with exogenous inputs (ARX), used to obtain damage sensitive features of a structure. To validate the performance of the proposed wireless monitoring and damage detection system, two near full scale single-story RC-frames, with and without brick wall system, are instrumented with the wireless monitoring system for real time damage detection during shaking table tests. White noise and seismic ground motion records are applied to the base of the structure using a shaking table. Pattern classification methods are then adopted to classify the structure as damaged or undamaged using time series coefficients as entities of a damage-sensitive feature vector. The demonstration of the damage detection methodology is shown to be capable of identifying damage using a wireless structural monitoring system. The accuracy and sensitivity of the MEMS-based wireless sensors employed are also verified through comparison to data recorded using a traditional wired monitoring system.

Design of radiation detection circuit for gamma column scanning (자동 감마 증류탑 검사 장치를 위한 방사선 계측장치 설계)

  • Kim, Jong-Beom;Jeong, Seong-Hui
    • Proceedings of the KIEE Conference
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    • 2003.11c
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    • pp.612-615
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    • 2003
  • In this paper, a design of radiation detector for gamma column scanner is introduced. Distillation column is important unit in Petro-chemical industries, and its on-line diagnose is very important. To get density profile measured by the radiation transmitted through column is well method for on-line diagnose as gamma scanning. For this purpose radiation detection circuit, radiation source and mechanical system for moving source and detector are required. Conventional radiation detection circuit for this application is sensitive to electric noise because of interface between the radiation circuit and the controller for mechanical system. The radiation detection system introduced here is using loop coil instead of slip ring to remove contact noise. Radiation detection system designed here for gamma scanning consist of BGO detector, high voltage circuit, PHA circuit and FSK modem. The BGO detector is used as radiation sensor, high voltage circuit and peak height analysis circuit is essential to process the signal generated from BGO detector. Micro controller convert measured data into ASCII data. FSK modem transmit ASCII data. Transmitted ASCH data is picked up in antenna coil and processed for combined function with mechanical system. This method gives good result by isolating the controlling circuit of mechanical system from radiation detecting circuit which is sensitive to noise.

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Simple and Rapid Detection of Potato leafroll virus by Reverse Transcription Loop-mediated Isothermal Amplification

  • Ju, Ho-Jong
    • The Plant Pathology Journal
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    • v.27 no.4
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    • pp.385-389
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    • 2011
  • A new reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for the Potato leafroll virus (PLRV) was developed and compared with conventional reverse transcription polymerase chain reaction (RT-PCR) to address its advantages over RTPCR. RT-LAMP primers were designed from the open reading frame 3 (ORF3) sequence of PLRV. The RT-LAMP reactions were conducted without or with a set of loop primers. By real-time monitoring using Turbimeter, the RT-LAMP (with loop primers) detects PLRV in less than 30 min, compared to 120 min of RT-PCR. By adding fluorescent reagent during the reaction, final products of the RT-LAMP were fluorescently visualized under UV light or could be differentiated by naked-eye inspection under normal light. The RT-LAMP was extremely sensitive, about 2000-fold more sensitive than RT-PCR. This study presents great potential of the RT-LAMP for diagnosis and PLRV epidemiology because RT-LAMP method is speedy, sensitive, inexpensive, and convenient.

Microscopic Detection of Urinary Tract Infection in Nepalese Patients

  • Dhakal, Bijaya-Kumar;Pokhrel, Bharat-Mani;Joohong Ahnn
    • Journal of Microbiology
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    • v.40 no.4
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    • pp.267-273
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    • 2002
  • Urinary tract infection (UTI) is one of the most common domiciliary and nosocomial bacterial infections prevalent in both males and females. UTI is diagnosed on the basis of clinical symptoms, microscopy and culture of urine. In order to evaluate the efficacy of microscopic detection for presumptive diagnosis of UTI we analyzed urine samples of Nepalese patients. We have conducted Gram staining and counting of pus cells, red blood cells (RBC) and epithelial cells. We observed that RBC and epithelial cell counts were not sensitive enough to be used for presumptive diagnosis of UTI. However, pus cell counts as well as Gram stain are sensitive and significant enough to presume UTI. When the Gram stain result was compared with the culture result, it was statistically significant. From this, we suggest that Gram stain of centrifuged urine is a very sensitive screening method to detect bacteriuria. In addition, we found that E. coli was the most predominant microorganism causing UTI and nitrofurantoin was the most effective antibiotic against the isolated urinary pathogens.

Ion-Sensitive Field Effect Transistor-Based Multienzyme Sensor for Alternative Detection of Mercury ions, Cyanide, and Pesticide

  • Vyacheslav, Volotovskky;Kim, Nam-Soo
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.373-377
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    • 2003
  • Various groups of industrial and agricultural pollutants (heavy metal ions, cyanides, and pesticides) can be detected by enzymes. Since heavy metal ions inhibit urease, cyanides inhibit peroxidase, organophosphorus and carbamate pesticides inhibit butyrylcholinesterase, these enzymes were co-immobilized into a bovine serum albumin gel on the surface of an ion-sensitive field effect transistor to create a bioprobe that is sensitive to the compounds mentioned above. The sensitivity of the present sensor towards KCN corresponded to $1\;\mu\textrm{M}$ with 1 min of incubation time. The detection limits for Hg(II) ions and the pesticide carbofuran were 0.1 and $0.5\;\mu\textrm{M}$, respectively, when a 10 min sensor incubation time in contaminated samples was chosen. The total time for determining the concentrations of all species mentioned did not exceed 20 min.

Sensitive, Accurate PCR Assays for Detecting Harmful Dinoflagellate Cochlodinium polykrikoides Using a Specific Oligonucleotide Primer Set

  • Kim Chang-Hoon;Park Gi-Hong;Kim Keun-Yong
    • Fisheries and Aquatic Sciences
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    • v.7 no.3
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    • pp.122-129
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    • 2004
  • Harmful Cochlodinium polykrikoides is a notorious harmful algal bloom (HAB) species that is causing mass mortality of farmed fish along the Korean coast with increasing frequency. We analyzed the sequence of the large subunit (LSD) rDNA D1-D3 region of C. polykrikoides and conducted phylogenetic analyses using Bayesian inference of phylogeny and the maximum likelihood method. The molecular phylogeny showed that C. polykrikoides had the genetic relationship to Amphidinium and Gymnodinium species supported only by the relatively high posterior probabilities of Bayesian inference. Based on the LSU rDNA sequence data of diverse dinoflagellate taxa, we designed the C. polykrikoides-specific PCR primer set, CPOLY01 and CPOLY02 and developed PCR detection assays for its sensitive, accurate HAB monitoring. CPOLY01 and CPOLY02 specifically amplified C. polykrikoides and did not cross-react with any dinoflagellates tested in this study or environmental water samples. The effective annealing temperature $(T_{p})$ of CPOLY01 and CPOLY02 was $67^{\circ}C$. At this temperature, the conventional and nested PCR assays were sensitive over a wide range of C. polykrikoides cell numbers with detection limits of 0.05 and 0.0001 cells/reaction, respectively.

Comparision of TTC-II and disk assay method for the detection of antibiotic residues in milk (우유내 잔류물질 검사방법인 TTC-II법 및 Disk assay법의 비교시험)

  • 이정아;이은미;이성해;도재철;박영구
    • Korean Journal of Veterinary Service
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    • v.22 no.1
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    • pp.93-101
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    • 1999
  • This test process on screening method for the detection of residual antibiotics in milk is simple, economic, sensitive to residual antibiotics and was given approval international organs. Thus, this study was carried out the comparison of Disk assay method and TTC-II method for sensitivity and minimum detectable range of antibiotics in raw milk. The results of this study was summarized as follows ; 1. The number of samples requested for treatment of mastitis was 198 samples. Comparison or analytical results among the methods of TTC-II, disk assay and Delve sp was that TTC-II 37 samples(18.6% ), Disk assay 125samples(63.1%), Delve SP 130 samples(65.7% ) reacted positively. Conformity rate of Delve SP and Disk assay was 70%. 2. Detectable limits of disk assay method in some antibiotics were more sensitive than those of official method(0.05-0.0025ppm in the $\beta$-lactams, 1ppm in two aminoglycoside, 0.2 ppm in one tetracycline, similar in one macrolide) 3. For sensitivity of residual sulfonamides TTC-II was much more sensitive than disk assay. Detectable limits of sulfamethazine and sulfadimethoxine were 30 to 50ppm levels. 4. The best medium preservation period is 1-2 days. 5. Concentration of brome cresol purple related to resistance for B stearothermophilus culture was 24ppm/ml. These results show that disk assay method for screening detection of antibiotics residuces in milk is worthy of use.

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