• Title/Summary/Keyword: method detection limit

Search Result 1,354, Processing Time 0.023 seconds

Simultaneous Detection of Four Foodborne Viruses in Food Samples Using a One-Step Multiplex Reverse Transcription PCR

  • Lee, Shin-Young;Kim, Mi-Ju;Kim, Hyun-Joong;Jeong, KwangCheol Casey;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
    • /
    • v.28 no.2
    • /
    • pp.210-217
    • /
    • 2018
  • A one-step multiplex reverse transcription PCR (RT-PCR) method comprising six primer sets (for the detection of norovirus GI and GII, hepatitis A virus, rotavirus, and astrovirus) was developed to simultaneously detect four kinds of pathogenic viruses. The size of the PCR products for norovirus GI and GII, hepatitis A virus (VP3/VP1 and P2A regions), rotavirus, and astrovirus were 330, 164, 244, 198, 629, and 449 bp, respectively. The RT-PCR with the six primer sets showed specificity for the pathogenic viruses. The detection limit of the developed multiplex RT-PCR, as evaluated using serially diluted viral RNAs, was comparable to that of one-step single RT-PCR. Moreover, this multiplex RT-PCR was evaluated using food samples such as water, oysters, lettuce, and vegetable product. These food samples were artificially spiked with the four kinds of viruses in diverse combinations, and the spiked viruses in all food samples were detected successfully.

Species-Specific Duplex PCR for Detecting the Important Fish Pathogens Vibrio anguillarum and Edwardsiella tarda

  • Jo, Geon-A;Kwon, Sae-Bom;Kim, Na-Kyeong;Hossain, Muhammad Tofazzal;Kim, Yu-Ri;Kim, Eun-Young;Kong, In-Soo
    • Fisheries and Aquatic Sciences
    • /
    • v.16 no.4
    • /
    • pp.273-277
    • /
    • 2013
  • Vibriosis caused by Vibrio anguillarum and edwardsiellosis caused by Edwardsiella tarda are septicemic diseases of many commercially important freshwater and marine fishes, and threaten the aquaculture industry in Korea. Early diagnosis and accurate identification of these two bacterial species could help to prevent these diseases and minimize the damage to cultured marine species. This study designed a duplex polymerase chain reaction (PCR) method for the simultaneous detection of two major fish pathogens: V. anguillarum and E. tarda. Each pair of oligonucleotide primers exclusively amplified the target groEL gene of the specific microorganism. Twenty-two Vibrio and ten non-Vibrio enteric species were used to check the specificity of the primers, which were found to be highly specific for the target species, even among closely related species. The detection limit was 400 pg for V. anguillarum and 4 ng for E. tarda when mixed purified DNA was used as the template. This assay showed high specificity and sensitivity in the simultaneous detection of V. anguillarum and E. tarda from artificially inoculated seawater and fish.

Analysis of Anions by Indirect Photometric Detection (I) (간접 분광 검출법에 의한 음이온의 분석(I))

  • Park, Man-Ki;Kim, Bak-Kwang;Park, Jeong-Hill;Kim, Kyoung-Ho;Lee, Mi-Yung;Jung, Jae-Eun
    • YAKHAK HOEJI
    • /
    • v.34 no.3
    • /
    • pp.215-218
    • /
    • 1990
  • An ion chromatographic method based on indirect photometric detection of UV transparent anions was developed. Separation of anion was accomplished on strong anion exchange column (Waters SAX) using UV detector at 254 nm. Among examined UV-active additives (Dns-H, Dns-glu, Dnsser, Dns-val), Dns-glu showed the highest sensitivity. Studies on the effects of the pH and ionic strength of eluent revealed that the increase of pH and ionic strength of the eluent decreased capacity factor. The best eluent for the separation of acetate, fluoride, chloride, nitrate and bromide was $1\;{\times}\;10^{-4}M$ Dns-glu in 5 mM phosphate buffer (pH 6.30). The detection limit of chloride ion was 2.1 ng in this condition.

  • PDF

Analysis of Saikosaponins by HPLC with Photoreduction Fluorescence Detection (광반응 HPLC를 이용한 시호 사포닌의 분석)

  • Shin, Young-Geun;Cho, Kyung-Hee;Kwon, Soo-Jin;Do, Young-Mi;Hwang, Gwi-Seo;Park, Jeong-Hill;Park, Man-Ki
    • YAKHAK HOEJI
    • /
    • v.40 no.1
    • /
    • pp.41-45
    • /
    • 1996
  • A high performance liquid chromatography using photoreduction fluorescence detection was described for the analysis of saikosaponins. Saikosaponins were separated on an $NH_2$ column using acetonitrile and aqueous 2-tert-butylanthraquinone(t-BAQ) as mobile phase. Column effluent was passed through a 40cm PTFE capillary tube coiled around a 10W UV lamp to reduce t-BAQ to a highly fluorescent dihydroxyanthracene derivative which was detected by a fluorescence detector. The optimal concentration of t-BAQ was found to be $6{\times}10^{-5}M$ and the optimal reaction time to be 2 seconds. The detection limit for saikosaponin a and d by this method was found to be about 280ng and 80ng. The dynamic linear range was over two orders and the correlation coefficient of the calibration curve of them was 0.998.

  • PDF

Real-Time Eye Detection and Tracking Under Various Light Conditions (다양한 조명하에서 실시간 눈 검출 및 추적)

  • 박호식;배철수
    • Journal of the Korea Institute of Information and Communication Engineering
    • /
    • v.8 no.2
    • /
    • pp.456-463
    • /
    • 2004
  • Non-intrusive methods based on active remote IR illumination for eye tracking is important for many applications of vision-based man-machine interaction. One problem that has plagued those methods is their sensitivity to lighting condition change. This tends to significantly limit their scope of application. In this paper, we present a new real-time eye detection and tacking methodology that works under variable and realistic lighting conditions. Based on combining the bright-Pupil effect resulted from IR light and the conventional appearance-based object recognition technique, our method can robustly track eyes when the pupils ale not very bright due to significant external illumination interferences. The appearance model is incorporated in both eyes detection and tacking via the use of support vector machine and the mean shift tracking. Additional improvement is achieved from modifying the image acquisition apparatus including the illuminator and the camera.

Urinary Biomarkers for the Noninvasive Detection of Gastric Cancer

  • Li, Dehong;Yan, Li;Lin, Fugui;Yuan, Xiumei;Yang, Xingwen;Yang, Xiaoyan;Wei, Lianhua;Yang, Yang;Lu, Yan
    • Journal of Gastric Cancer
    • /
    • v.22 no.4
    • /
    • pp.306-318
    • /
    • 2022
  • Gastric cancer (GC) is associated with high morbidity and mortality rates. Thus, early diagnosis is important to improve disease prognosis. Endoscopic assessment represents the most reliable imaging method for GC diagnosis; however, it is semi-invasive and costly and heavily depends on the skills of the endoscopist, which limit its clinical applicability. Therefore, the search for new sensitive biomarkers for the early detection of GC using noninvasive sampling collection methods has attracted much attention among scientists. Urine is considered an ideal biofluid, as it is readily accessible, less complex, and relatively stable than plasma and serum. Over the years, substantial progress has been made in screening for potential urinary biomarkers for GC. This review explores the possible applications and limitations of urinary biomarkers in GC detection and diagnosis.

The Improved Method for Precise Determination of Pu Isotope Ratio using MC-ICP-MS (다중검출기유도결합플라즈마질량분석기를 이용한 Pu 동위원소비 정밀 분석법)

  • Yim, Seong-A;Han, Eun-Mi;Chae, Jung-Seok;Yun, Ju-Young
    • Journal of Radiation Protection and Research
    • /
    • v.35 no.3
    • /
    • pp.117-123
    • /
    • 2010
  • Plutonium is by far the most important of the transuranic elements which have been released into the environment due to radio-toxicity and long term radiation effects on humans. And Pu isotope ratio ($^{240}Pu/^{239}Pu$) is of great interest because this ratio is used as a fingerprint for different sources. Mass spectrometry has been used as an useful atom counting technique with several advantages over decay counting techniques for the determination of Pu isotopes. It enables a determination of Pu isotope ratio in the environmental samples with a low detection limit and a short determination time. An ICP-MS is the representative mass spectrometry for Pu determination. In this study, the precision of $^{240}Pu/^{239}Pu$ isotope ratio was improved by using 4 multiple ion counters of MC-ICP-MS. The detection limit of $^{239}Pu$ and $^{240}Pu$ were $0.10\;fg\;ml^{-1}$ ($0.24\;{\mu}Bq\;ml^{-1}$), $0.12\;fg\;ml^{-1}$ ($0.97\;{\mu}Bq\;ml^{-1}$), respectively. The relative standard deviation of $^{240}Pu/^{239}Pu$ isotope ratio was less than 1 % in trace level. The various reference materials (seawater, soil and sediment) were analyzed to verify this method and their analytical results were in good agreement with the certified (or recommended value) value.

Site-directed Immobilization of Antibody onto Solid Surfaces for the Construction of Immunochip

  • Paek, Se-Hwan;Cho, Il-Hoon;Paek, Eui-Hwan;Lee, Haewon;Park, Jeong-Woo
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.9 no.2
    • /
    • pp.112-117
    • /
    • 2004
  • The performance of an immuno-analytical system can be assessed in terms of its analytical sensitivity, i.e., the detection limit of an analyte, which is determined by the amount of analyte molecules bound to the capture antibody that has been immobilized onto a solid surface. To increase the number of the binding complexes, we have investigated a site-directed immobilization of an antibody that has the ability to resolve a current problem associated with a random arrangement of the insolubilized immunoglobulin. The binding molecules were chemically reduced to produce thiol groups that were limited at the hinge region, and then, the reduced products were coupled to biotin. This biotinylated antibody was bound to a streptavidin-coated surface via the streptavidin-biotin reaction. This method can control the orientation of the antibody molecules present on a solid surface and also can significantly reduce the possibility of steric hindrance in the antigen-antibody reactions. In a two-site immunoassay, the introduction of the site-directly immobilized antibody as the capture enhanced the sensitivity of analyte detection approximately 10 times compared to that of the antibody randomly coupled to biotin. Such a novel approach would offer a protocol of antibody immobilization in order for the possibility of constructing a high performance immunochip.

Collagenolytic Activity Of Gingival Crevicular Fluid In Progressive Periodontitis (진행성 치주염에서의 치은열구액내 교원질분해효소 활성)

  • Chung, Hyun-Ju
    • Journal of Periodontal and Implant Science
    • /
    • v.26 no.1
    • /
    • pp.161-175
    • /
    • 1996
  • There were many reports that elevations in the levels of active and latent collagenase in gingival crevicular fluid(GCF) have been correlated positively with periodontal disease activity. To provide a simple diagnostic approach for testing GCF collagenolytic activity, the detection limit of enzyme activity was compared using radiofibril assay(Sodek et.al.1981) and spectrophotometric collagenolytic assay(Nethery et al. 1986). The detection limits of both assay for standard bacterial enzyme were similar and the radiofibril assay showed a little (1/2) lower detection limit for tad pole collagenase. To evaluate the relationship between periodontal tissue destruction and the collagenolytic activity, GCF was collected, and latent and active enzyme activities were measured by a spectrophotometric collagenolytic assay. Twelve subjects showing progressive lesions were selected according to the presence of immediate tissue destruction, frequent abscess formation, and increasing need for tooth extraction, and the absence of underlying systemic disease and previous antibiotic medication history within 6 months. Comparisons were made between sites with either: 1) inflammation with a previous history of progressive loss of periodontal tissue and bone support(2l progressive sites): 2) previous history of bone loss and periodontal destruction but now clinically stable(12 comparably stable sites); or 3) no loss of periodontal tissue and bone support(11 control sites including 5 gingivitis sites and 6 healthy sites). Active collagenase activity was the highest in the progressive sites and decreased in the order of the gingivitis sites, the stable sites, and the healthy sites. The total enzyme activity was $2{\sim}3$ fold higher in the progressive sites and the gingivitis sites, compared to the stable and the healthy sites. The ratio of active to total collagenolytic activity was twice in the progressive sites. Analysis of active collagenase level(5mU) and the ratio of active to total collagenolytic activity(0.8) as a diagnositic test indicates that these measurements have the sensitivity of 0.81 and 0.86, the specificity of 0.70 and 0.65, and the overall agreement of 0.75 and 0.73, respectively. Thus, this method has significant merits as a diagnostic tool to determine wherher the site is in a state of remission or progression.

  • PDF

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
    • /
    • v.38 no.5
    • /
    • pp.347-355
    • /
    • 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.