• 제목/요약/키워드: method detection limit

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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
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    • 제28권2호
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    • pp.210-217
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    • 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
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    • 제16권4호
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    • pp.273-277
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    • 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.

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

  • 박만기;김박광;박정일;김경호;이미영;정재은
    • 약학회지
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    • 제34권3호
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    • pp.215-218
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    • 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.

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

  • 신영근;조경희;권수진;도영미;황귀서;박정일;박만기
    • 약학회지
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    • 제40권1호
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    • pp.41-45
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    • 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.

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

  • 박호식;배철수
    • 한국정보통신학회논문지
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    • 제8권2호
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    • pp.456-463
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    • 2004
  • 본 논문에서는 다양한 조명하에서 실시간으로 눈을 검출하고 추적하는 새로운 방법을 제안하고자 한다. 기존의 능동적 적외선을 이용한 눈 검출 및 추적 방법은 외부의 조명에 매우 민감하게 반응하는 문제점을 가지고 있으므로, 본 논문에서는 적외선 조명을 이용한 밝은 동공 효과와 전형적인 외형을 기반으로 한 사물 인식 기술을 결합하여 외부 조명의 간섭으로 밝은 동공 효과가 나타나지 않는 경우에도 견실하게 눈을 검출하고 추적 할 수 있는 방법을 제안한다. 눈 검출과 추적을 위해 SVM과 평균 이동 추적방법을 사용하였고, 적외선 조명과 카메라를 포함한 영상 획득 장치를 구성하여 제안된 방법이 효율적으로 다양한 조명하에서 눈 검출과 추적을 할 수 있음을 보여 주었다.

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
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    • 제22권4호
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    • pp.306-318
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    • 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.

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

  • 임성아;한은미;채정석;윤주용
    • Journal of Radiation Protection and Research
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    • 제35권3호
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    • pp.117-123
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    • 2010
  • Pu 동위원소는 기원에 따라 각기 다른 $^{240}Pu/^{239}Pu$ 동위원소비를 나타내는 특성이 있다. 질량분석법은 극미량 장반감기핵종 분석에 매우 유용한 방법으로서 검출기 특성상 낮은 검출한계를 갖고 있으며, 소량의 시료를 이용하여 짧은 시간에 측정을 할 수 있다는 장점을 갖고 있으므로 Pu을 비롯한 장반감기 핵종 분석에 유용하다. 다중검출기유도결합플라즈마질량분석기(Multicollector ICP-MS)는 다중검출시스템을 이용하여 분석하고자 하는 여러 핵종의 동시분석이 가능하며, 특별히 본 연구에서는 극미량 핵종의 검출이 가능한 multiple ion counter (MIC)를 사용하여 Pu 동위원소 분석에 적용하였다. 본 연구방법의 검출한계(detection limit)는 최적화된 조건에서 $^{239}Pu$$^{240}Pu$ 각각 $0.10\;fg\;ml^{-1}$ ($0.24\;{\mu}Bq\;ml^{-1}$), $0.12\;fg\;ml^{-1}$ ($0.97\;{\mu}Bq\;ml^{-1}$)이었으며, 240Pu 기준으로 약 200 cps 정도의 매우 낮은 농도수준에서 1%이하의 $^{240}Pu/^{239}Pu$ 동위원소비 계측편차를 나타내었다. 본 연구방법은 표준용액 및 다양한 매트릭스의 표준물질 분석에 적용함으로써 검증 확인하였다.

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
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    • 제9권2호
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    • pp.112-117
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    • 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.

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

  • 정현주
    • Journal of Periodontal and Implant Science
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    • 제26권1호
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    • pp.161-175
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    • 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.

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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
    • 한국식품위생안전성학회지
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    • 제38권5호
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    • pp.347-355
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    • 2023
  • 본 연구에서는 시가독소 생성 대장균(STEC)을 검출하기 위해 식품공전의 polymerase chain reaction (PCR)검사법과 loop-mediated isothermal amplification (LAMP)를 비교하였다. PCR 및 LAMP의 검출 한계(LOD) 및 정량화 한계(LOQ), 민감도, 특이성 및 효율성을 평가하기 위해 다양한 식품에 STEC를 접종하였다. LOD는 PCR의 경우 104 CFU/mL 이하, LAMP의 경우 103 CFU/mL 이하로 측정되었다. LOQ 값은 PCR과 LAMP 간에 차이가 없었다. 그러나 4가지 식품군에서 민감도는 양념육이 최대 11.1%, 간소고기가 최소 8.1% 차이가 났다. LAMP는 네 가지 음식 유형 모두에 대해 높은 민감도와 100% 특이도를 보였다. 따라서 LAMP는 식품 유형에 따라 검출률이 비슷하고 특이도와 민감도가 식품공전 PCR보다 우수하기 때문에 STEC에 대한 신뢰할 수 있는 분자 검출 방법이다.