• Title/Summary/Keyword: Limit of detection(LOD)

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Comparative Evaluation of Loop-Mediated Isothermal Amplification (LAMP) and Conventional PCR for Detection of Shiga-Toxin-Producing Escherichia coli (STEC) in Various Food Products

  • Hyejin Jang;Yong Sun Cho
    • Journal of Food Hygiene and Safety
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    • v.38 no.5
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    • pp.347-355
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    • 2023
  • In this study, polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) were compared in terms of their ability to detect shiga-toxin-producing Escherichia coli (STEC). Various foods were artificially inoculated with STEC to evaluate the limit of detection (LOD), limit of quantification (LOQ), sensitivity, specificity, and efficiency of PCR and LAMP. The LODs were ≤104 and ≤103 CFU/mL for PCR and LAMP, respectively. The LOQs did not differ between PCR and LAMP. However, of the four considered food types, the sensitivities differed by a maximum of 11.1% for seasoned meat and by a minimum of 8.1% for ground beef. LAMP had higher sensitivity than that of PCR and 100% specificity for all four food types. Therefore, LAMP is a reliable molecular method for detecting STEC as comparable to PCR assay, and its specificity and sensitivity are superior to those of PCR, depending on the food type.

Development and Characterization of Finger-type PIN Photodiode for Fluorescence Detection of RNA (RNA 형광 검출을 위한 Finger형 PIN 광다이오드의 제작 및 평가)

  • Kim, Ju-Hwan;Oh, Myung-Hwan;Ju, Byeong-Kwon
    • Journal of Sensor Science and Technology
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    • v.13 no.2
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    • pp.85-89
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    • 2004
  • This paper represents the development of high sensitivity photo-sensor for the fluorescence detection in the integrated biological analysis system. The finger-type PIN photodiodes were fabricated as the photo-sensor, and had a high sensitivity ($I_{light}/I_{dark}$ = 8720). The interference filter consisted of $TiO_{2}$ and $SiO_{2}$ was directly deposited on the photodiodes. Deposited filter with 95.5% reflection under 532 nm and 98% transmission over 580 nm exceedingly decreased the magnitude of background signal in the detection. The PDMS micro-fluidic channels are bonded on the photodiode by $O_{2}$ plasma treatment. The detection current was proportional to two primary parameters (light intensity, concentration), and the on-chip detection system could detect fluorescence signals down to 100 nM concentration (LOD = Limit of detection of rhodamine).

Simultaneous Analysis of 17 Organophosphorous Pesticides in Blood by Automated Head Space-SPME GC/MS (HS-SPME-GC/MS에 의한 혈액중 17종 유기인계 농약의 동시분석법)

  • Rhee, Jong-Sook;Jung, Jin-Mi;Lee, Han-Sun;Yeom, Hye-Sun;Lee, Sang-Ki;Park, Yoo-Sin;Chung, Hee-Sun
    • YAKHAK HOEJI
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    • v.54 no.6
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    • pp.429-440
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    • 2010
  • HS-SPME-GC/MS was studied and optimized for the determination of 17 orgarnophosphorous pesiticides (OPPs: chlorpyrifos, chlorpyrifos-methyl, demeton-s-methyl, diazinon, dimethoate, EPN, fenitrothion, fenthion, malathion, methidathion, monocrotophos, parathion, phenthoate, phosphamidon, sulfotep, terbufos, triazophos) in blood. Optimum SPME parameters were selected: choice of SPME fiber (85 ${\mu}m$ polyacrylate), pH effect (0.5 N HCl), salt effect ($Na_2SO_4$, 0.2 g; 20%), headspace incubation temperature ($80^{\circ}C$), headspace incubation time (1 min), headspace adsorption time (30 min) and GC desorption time (2 min). These parameters were optimized using HS-SPME autosampler coupled with gas chromatography-mass spectrometry (GC-MS). Method validation was carried out in terms of linearity, limit of detection (LOD), limit of quantitation (LOQ) and recovery in blood. The assay was linear over 0.5~5.0 mg/l ($r^2$=0.955~1.000). Limit of detection (LOD) and limit of quantitation (LOQ) in blood were determined 0.03~0.3 mg/l (S/N=3) and 0.1~1.1 mg/l (S/N=10), respectively. Relative recovery with 0.5, 1 and 5 mg/l (in blood) were 90.8%, 98.5% and 94.1%, respectively. This method will be applied to the determination of the orgarnophosphorous pesticides in postmortem blood. The proposed protocol can be an attractive alternative to be used in routine toxicological analysis.

A Modified Quantum Dot-Based Dot Blot Assay for Rapid Detection of Fish Pathogen Vibrio anguillarum

  • Zhang, Yang;Xiao, Jingfan;Wang, Qiyao;Zhang, Yuanxing
    • Journal of Microbiology and Biotechnology
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    • v.26 no.8
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    • pp.1457-1463
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    • 2016
  • Vibrio anguillarum, a devastating pathogen causing vibriosis among marine fish, is prevailing in worldwide fishery industries and accounts for grievous economic losses. Therefore, a rapid on-site detection and diagnostic technique for this pathogen is in urgent need. In this study, two mouse monoclonal antibodies (MAbs) against V. anguillarum, 6B3-C5 and 8G3-B5, were generated by using hybridoma technology and their isotypes were characterized. MAb 6B3-C5 was chosen as the detector antibody and conjugated with quantum dots. Based on MAb 6B3-C5 labeled with quantum dots, a modified dot blot assay was developed for the on-site determination of V. anguillarum. It was found that the method had no cross-reactivity with other than V. anguillarum bacteria. The detection limit (LOD) for V. anguillarum was 1 × 103 CFU/ml in cultured bacterial suspension samples, which was a 100-fold higher sensitivity than the reported colloidal gold immunochromatographic test strip. When V. anguillarum was mixed with turbot tissue homogenates, the LOD was 1 × 103 CFU/ml, suggesting that tissue homogenates did not influence the detection capabilities. Preenrichment with the tissue homogenates for 12 h could raise the LOD up to 1 × 102 CFU/ml, confirming the reliability of the method.

Liquid electrochemical sensors using carbon nanotube film (Carbon Nanotube Film을 이용한 액체 전기화학 센서)

  • Noh, Jaeha;An, Sangsu;Lee, Changhan;Lee, Sangtae;Lee, Moonjin;Seo, Dongmin;Chang, Jiho
    • Journal of Sensor Science and Technology
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    • v.30 no.4
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    • pp.255-260
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    • 2021
  • We studied electrochemical sensors using printed carbon nanotube (CNT) film on a polyethylene terephthalate (PET) substrate. Multiwalled CNT films were printed on a PET substrate to study its feasibility as hazardous and noxious substances (HNS) detection sensor. The printed CNT film (PCF) with a 50 ㎛ thickness exhibited a specific resistance of 230 ohm. To determine the optimum sensor structure, a resistance-type PCF sensor (R-type PCF sensor) and a conductive-type PCF sensor (C-type PCF sensor) were fabricated and compared using diluted NH3 droplets with various concentrations. The response magnitude, response time, sensitivity, linearity, and limit of detection (LOD) were compared, and it was concluded that the C-type PCF sensor exhibited superior performance. By applying a C-Type PCF sensor, we confirmed the detection performance of 12 types of floating HNS and the response of the sensor with selectivity according to the degree of polarity.

Validation and Uncertainty Evaluation of an Optimized Analytical Method Using HPLC Applied to Canthaxanthin, a Food Colorant (식품 색소 Canthaxanthin의 HPLC 최적 분석법 확인 및 타당성과 측정불확도 평가)

  • Suh, Hee-Jae;Kim, Kyung-Su;Hong, Mi-Na;Lee, Chan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.3
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    • pp.342-351
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    • 2016
  • This study was carried out to develop an optimized analytical method using high-performance liquid chromatography (HPLC) applied to canthaxanthin, which is not yet designated as a food colorant in Korea, as well as to perform validation and uncertainty evaluation of this method. Official methods of AOAC, UK, and Japan with HPLC-UV detection were evaluated for the analysis of canthaxanthin by comparison of linearity, resolution, selectivity, limit of detection (LOD), limit of quantitation (LOQ), accuracy, precision, recovery, inter-laboratory tests, and uncertainty measurement. The calibration curves showed high linearity with an $R_2$ value of over 0.999 for canthaxanthin standard solutions in all three official methods. The official method of Japan exhibited the best results in terms of resolution and selectivity, including the lowest LOD and LOQ. The average coefficients of variation were calculated as less than five of three institutes with a precision value less than 1, accuracy near 100%, and recovery ratio between $100{\pm}10%$. The expanded uncertainty for canthaxanthin was estimated to be $39.5{\pm}5.29mg/kg$ (95% confidence level, k=2), and the uncertainty of measurement was 13.4%. In this study, official methods of canthaxanthin were compared and the validities verified. The results will be further applied to establish an authorized analytical method for canthaxanthin in Korea.

Evaluation of commercial immunochromatography test kits for diagnosing canine parvovirus

  • Lee-Sang Hyeon;Dong-Kun Yang;Eun-Ju Kim;Yu-Ri Park;Hye Jeong Lee;Bang-Hun Hyun
    • Korean Journal of Veterinary Research
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    • v.63 no.2
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    • pp.19.1-19.6
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    • 2023
  • Rapid immunochromatography test (RICT) kits are commonly used for the diagnosis of canine parvovirus (CPV) because of their rapid turnaround time, simplicity, and ease of use. However, the potential for cross-reactivity and low sensitivity can yield false-positive or false-negative results. There are 4 genotypes of CPV. Therefore, evaluating the performance and reliability of RICT kits for CPV detection is essential to ensure accurate diagnosis for appropriate treatment. In this study, we evaluated the performance of commercial RICT kits in the diagnosis of all CPV genotypes. The cross-reactivity of 6 commercial RICT kits was evaluated using 8 dog-related viruses and 4 bacterial strains. The limit of detection (LOD) was measured for the 4 genotypes of CPV and feline panleukopenia virus. The tested kits showed no cross-reactivity with the 8 dog-related viruses or 4 bacteria. Most RICT kits showed strong positive results for CPV-2 variants (CPV-2a, CPV-2b, and CPV-2c). However, the 2 kits produced negative results for CPV-2 or CPV-2b at a titer of 105 FAID50/mL, which may result in inaccurate diagnoses. Therefore, some kits need to improve their LOD by increasing their binding efficiency to detect all CPV genotypes.

Comparison of Sampling and Analytical Methods for Determining Airborne Hexavalent Chromium -Limit of Detection, Accuracy and Precision of Analytical Procedures (공기중 6가 크롬 측정 방법 비교 -검출한계, 정확도 및 정밀도-)

  • 신용철;이병규;이지태
    • Journal of Korean Society for Atmospheric Environment
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    • v.18 no.1
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    • pp.39-49
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    • 2002
  • In this study, limits of detection (LOD), accuracy and precision of four sampling/ analytical methods were evaluated and compared for the determination of airborne hexavalent chromium, Cr (VI). The methods include : (1) a combination of the National Institute for Occupational Safety and Health (NIOSH) Method 7600/U. S. Environmental Protection Agency (EPA) Method 218.6 (NIOSH/EPA Method) proposed by Shin and Paik, 2) two impinger methods using 2% NaOH/3% Na$_2$CO$_3$. (3) same as (2) but with 0.02 N NaHCO$_3$absorbing solution, and (4) the Occupational Safety and Health (OSHA) Method ID-215. An ion chromatograph/visible absorbance detector was used for the analysis of Cr (VI) in sample solution. Limit of detection (LOD) , analytical accuracy, and precision were also tested using Cr (VI) spike samples. Recoveries (as index of accuracy) and coefficient of variation (CV) (as a index of precision) were determined. Two-way ANOVA and Turkey's test were performed to test the significance in differences among recoveries and CVs of the methods. In all the methods, the peaks of Cr (VI) were separated sharply on chromatograms and exhibited a strong linearity with Cr (VI) concentrations in solution. The correlation coefficients of calibration curves typically ranged from 0.9997 to 0.9999, and the analytical LODs from 0.025 to 0.1$\mu\textrm{g}$/sample. All the method had good sensitivities and linearities between Cr (VI) levels and peak areas. The accuracies (% mean recoveries) of the methods ranged from 80.1 to 104.2%, while the precisions (pooled coefficient of variation) ranged from 3.16 to 4.43%. The impinger methods showed higher recoveries ( > 95%) than those of the PVC filter methods (the OSHA Method and the NIOSH/EPA Method). It was assumed that Cr (VI) on PVC filter was exposed to air and reduced to trivalent chromium, Cr (III), whereas it was stabilized in alkali solution contained in impinger. Thus, a special treatment of Cr (VI) samples collected on PVC filters may be required.

In-situ measurement of Ce concentration in high-temperature molten salts using acoustic-assisted laser-induced breakdown spectroscopy with gas protective layer

  • Yunu Lee;Seokjoo Yoon;Nayoung Kim;Dokyu Kang;Hyeongbin Kim;Wonseok Yang;Milos Burger;Igor Jovanovic;Sungyeol Choi
    • Nuclear Engineering and Technology
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    • v.54 no.12
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    • pp.4431-4440
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    • 2022
  • An advanced nuclear reactor based on molten salts including a molten salt reactor and pyroprocessing needs a sensitive monitoring system suitable for operation in harsh environments with limited access. Multi-element detection is challenging with the conventional technologies that are compatible with the in-situ operation; hence laser-induced breakdown spectroscopy (LIBS) has been investigated as a potential alternative. However, limited precision is a chronic problem with LIBS. We increased the precision of LIBS under high temperature by protecting optics using a gas protective layer and correcting for shotto-shot variance and lens-to-sample distance using a laser-induced acoustic signal. This study investigates cerium as a surrogate for uranium and corrosion products for simulating corrosive environments in LiCl-KCl. While the un-corrected limit of detection (LOD) range is 425-513 ppm, the acoustic-corrected LOD range is 360-397 ppm. The typical cerium concentrations in pyroprocessing are about two orders of magnitude higher than the LOD found in this study. A LIBS monitoring system that adopts these methods could have a significant impact on the ability to monitor and provide early detection of the transient behavior of salt composition in advanced molten salt-based nuclear reactors.

Validation and measurement uncertainty of HPLC-UV method for quercetin quantification in various foods

  • Seo, Eunbin;Lim, Suji;Yun, Choong-In;Shin, Jae-Wook;Kim, Young-Jun
    • Korean Journal of Food Science and Technology
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    • v.53 no.6
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    • pp.682-687
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    • 2021
  • The purpose of this study was to validate a high-performance liquid chromatography (HPLC) method for the quantitative analysis of quercetin in various foods. The method was based on HPLC-UV (360 nm). The method was validated using candy, beverage, and sausage which were tested for specificity, linearity, limit of detection (LOD), limit of quantification (LOQ), precision, and accuracy, and the measurement uncertainty was assessed. Matrix-matched calibration was also applied. The calibration curves (0.5-50 mg/L) showed good linearity (r2≥0.9998). LOD and LOQ ranged from 0.15 to 0.31 mg/kg and from 0.44 to 0.93 mg/kg, respectively. The average accuracy and precision at 0.5, 2.5, and 10 mg/kg ranged from 84.3 to 102.0% and 0.7 to 3.0 relative standard deviation (RSD%), respectively. This study confirmed the applicability of the proposed method by applying it to commercial products, such as teas and beverages. Thus, the proposed analytical method is suitable for quantifying quercetin in various foods.