• 제목/요약/키워드: Diagnostic kit

검색결과 162건 처리시간 0.029초

혈청 유리형 전립선항원 (free PSA) 측정을 위한 효소면역측정법의 개발 (Development of Enzyme Immuno Assay for Analysis of Free Prostate Specific Antigen in Serum)

  • Kyung-Ok Lee;Kyung-In Kim;Kyu-Pum Lee
    • 대한의생명과학회지
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    • 제3권2호
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    • pp.107-114
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    • 1997
  • 전립선특이 항원 (PSA: Prostate Specific Antigen)은 전립선 세포에서 분비되는 단백질로서 장기 특이성이 높아 전립선암의 조기진단 및 치료의 예후를 판단하기 위해 임상에서 널리 사용되고 있는 혈청 종양표지검사의 대상항원이다. 그러나 혈청 PSA치는 전립선암 뿐 아니라 양성 전립선질환인 전립선비대증 (BPH), 전립선 경색, 전립선 염증 등에서도 상승될 수 있다. 혈청 내의 PSA는 여러 가지 분자형태로 이루어져 있으며 대표적으로 ${alpha}_1$-antichymotrypsin과 결합된 결합형 PSA(PSA-ACT)와 유리형 PSA (free PSA)로 나눌 수 있다. 전립선 암 환자의 혈청에는 결합형 PSA가 95% 이상으로 매우 높고 유리형 PSA의 분포는 매우 작은 반면 전립성비대증에서는 유리형 PSA의 농도가 높아지므로, 결합형 PSA와 동시에 유리형 PSA를 측정하면 종양표지검사로서의 특이성과 예민도를 높일 수 있다는 보고가 있어, 최근 혈청내의 유리형 PSA의 측정의 중요성이 임상적으로 대두되고 있다. 이에 본 연구에서는 sandwich 원리로 유리형 PSA 효소면역 측정법 (EIA) kit를 개발하고 그 유용성을 검토한 결과, 고, 저 농도에서의 일내, 일간 변이 계수 (CV)는 4% 이하였으며 상품화된 free PSA kit와 비교하였을 때 두 방법간의 상관계수는 0.9965으로 매우 양호하였다. 또한 농도가 높은 세 환자의 검체를 희석하여 직선성 검사를 하였을 때 상관계수가 모두 0.995이상으로 나타났다. 또한 microplate 법에서 문제될 수 있는 hook effect는 유리형 PSA농도가 40 ng/mL까지 나타나지 않았으며, 98.9%~104.1%의 회수율을 나타내었다. 따라서 본 연구에서 개발한 면역측정법 kit는 혈청 중의 유리형 PSA를 정확하고 간편하게 분석할 수 있으므로 임상 실험실에서 전립선암이나 전립선질환의 진단 및 치료효과의 판정에 유용하게 사용될 수 있을 것으로 기대된다.

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Comparative analysis of serological tests and fecal detection in the diagnosis of Mycobacterium avium subspecies paratuberculosis infection

  • Park, Hong-Tae;Ha, Seungmin;Park, Hyun-Eui;Shim, Soojin;Hur, Tai Young;Yoo, Han Sang
    • 대한수의학회지
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    • 제60권3호
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    • pp.117-122
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    • 2020
  • Johne's disease (JD) caused by Mycobacterium avium subspecies paratuberculosis (MAP) is a chronic, wasting infectious disease in ruminants that causes enormous economic losses to the dairy and beef cattle industries. The most effective way to eradicate JD is to detect infected individuals as early as possible and remove them from the herd. However, it is difficult to detect infected individuals early with the currently using diagnostic methods. Two serological diagnostic kits commercially used worldwide and a fecal detection test were compared using 298 serum samples and feces of cattle in this study to present an efficient diagnostic method. Although there was a high correlation between the 2 serological diagnostic kits (R2 = 0.7473), kit A showed a higher serological positive rate. However, the correlation between fecal tests and serological diagnosis was very low. MAP was also detected in fecal tests in many serologically negative individuals. In the periodical diagnosis of JD, MAP was detected in the feces of only cows with the higher antibody titer to MAP. These results suggest that for effective eradication of JD, early detection of infected individuals by fecal tests together with the serological tests currently in use and by removal of infected individuals are needed.

TNF-${\alpha}$ Up-regulated the Expression of HuR, a Prognostic Marker for Ovarian Cancer and Hu Syndrome, in BJAB Cells

  • Lee, Kyung-Yeol
    • IMMUNE NETWORK
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    • 제4권3호
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    • pp.184-189
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    • 2004
  • Background: Hu syndrome, a neurological disorder, is characterized by the remote effect of small cell lung cancer on the neural degeneration. The suspicious effectors for this disease are anti-Hu autoantibodies or Hu-related CD8+ T lymphocytes. Interestingly, the same effectors have been suggested to act against tumor growth and this phenomenon may represent natural tumor immunity. For these diagnostic and therapeutic reasons, the demand for antibodies against Hu protein is rapidly growing. Methods: Polyclonal and monoclonal antibodies were generated using recombinant HuR protein. Western blot analyses were performed to check the specificity of generated antibodies using various recombinant proteins and cell lysates. Extracellular stimuli for HuR expression had been searched and HuR-associated proteins were isolated from polysome lysates and then separated in a 2-dimensional gel. Results: Polyclonal and monoclonal antibodies against HuR protein were generated and these antibodies showed HuR specificity. Antibodies were also useful to detect and immunoprecipitate endogenous HuR protein in Jurkat and BJAB. This report also revealed that TNF-${\alpha}$ treatment in BJAB up-regulated HuR expression. Lastly, protein profile in HuR-associated mRNAprotein complexes was mapped by 2-dimensional gel electrophoresis. Conclusion: This study reported that new antibodies against HuR protein were successfully generated. Currently, project to develop a diagnostic kit is in process. Also, this report showed that TNF-${\alpha}$ up-regulated HuR expression in BJAB and protein profile associated with HuR protein was mapped.

ELISA를 이용한 돼지 옴 (Sarcoptes scabiei var. suis) 감염증의 진단 (Diagnosis of swine sarcoptic mange(Sarcoptes scabiei var. suis) by ELISA)

  • 지차호;이삼선;장래훈
    • 대한수의학회지
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    • 제41권4호
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    • pp.565-570
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    • 2001
  • The diagnosis of swine sarcoptic mange (Sarcoptes scabiei var. suis) was investigated by ELISA in order to replace current diagnostic methods such as skin scraping, scratching index, or lesion score of dermatitis. The current methods need many efforts and much times and cost much. They can not handle many samples simultaneously. Therefore, in this research we developed ELISA that can handle many samples at a time. The antigens of swine sarcoptic mite were isolated and examined by 12.5% SDS-PAGE and silver staining. The antigenicity of antigen was confirmed by Western blotting using the swine from the artificailly-infested swine with swine sarcoptic mite. The optical density (OD) values of the artificailly-infested positive sera and the naturally-infested positive sera of sows were measured and read in order to confirm the stability of antigens, the reproducibility and validity (sensitivity and specificity) of the manufactured ELISA of swine sarcoptic antigens. In above results, the developed ELISA would be possible to use the diagnostic tool of sarcoptic mange if OD values of piglets, fattening pigs and sows are interpreted reasonably and classified as mange-free and infested.

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생명정보학 기반 H5N1에 특이적인 진단키트 개발을 위한 epitope 선별 (Selection of epitope for development of H5N1 specific diagnostic kit based on bioinformatics)

  • 이인성;김학용
    • 한국콘텐츠학회:학술대회논문집
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    • 한국콘텐츠학회 2014년도 추계 종합학술대회 논문집
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    • pp.57-58
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    • 2014
  • 인플루엔자 A 바이러스의 아형인 H5N1은 고병원성으로 조류 독감을 일으킨다. H5N1 바이러스는 원래 조류끼리만 감염되는 독감이고, 사람에게는 전염되지 않는다고 알려져 있었으나, 2003년에 베트남과 중국을 시작으로 현재까지 168명의 사망자가 기록되고 있다. 그러나 현재 시판되고 있는 진단키트(Rapid diagnostic kits)들은 H5N1 에 특이적인 것이 아니라 influenza A virus 모두를 진단한다. 따라서 influenza 감염여부는 확인 할 수 있지만, 이것이 H5N1 인지는 확인 할 수가 없다. H5N1은 전염성이 강하기 때문에 빠르게 진단하여 감염조류를 살 처분 하여야 더 많은 경제적 손실을 줄일 수 있다. 따라서 H5N1 에만 특이적인 epitope를 네트워크 기반으로 예측하여 진단제에 응용할 수 있도록 하고자 한다. 각 서열 정보는 Openflu (http://openflu. vital-it.ch/browse.php)에서 얻었다. H5N1은 H1N1에서 유래되었기 때문에 두 subtype의 차이점을 알아보고자 TCOFFEE에서 multiple sequence alignment를 수행한 결과 N-terminal 부분이 상이하였다. 상이한 H5N1의 N-terminal 부분이 H5N1 virus에 감염된 모든 host에서 존재하는지 알아보기 위해 host가 사람인 경우와 조류인 경우를 TCOFFEE에서 alignment 하였다. 그 결과 H5N1의 N-terminal 부분은 사람과 조류에서 보존적이었다. 따라서 H5N1의 N-terminal이 다른 subtype과 유사하지 않고 H5에만 특이적이기 때문에 진단키트 제작을 위한 epitope로 사용할 수 있을 것으로 기대된다.

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Characterization of the Salmonella typhi Outer Membrane Protein C

  • Toobak, Hoda;Rasooli, Iraj;Gargari, Seyed Latif Mousavi;Jahangiri, Abolfazl;Nadoushan, Mohammadreza Jalali;Owlia, Parviz;Astaneh, Shakiba Darvish Alipour
    • 한국미생물·생명공학회지
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    • 제41권1호
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    • pp.128-134
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    • 2013
  • Salmonella enterica serovar typhi, a Gram-negative food-borne pathogen, causes typhoid fever in humans. OmpC is an outer membrane porin of S. typhi expressed throughout the infection period. OmpC is potentially an attractive antigen for multivalent vaccines and diagnostic kit designs. In this study we combined in silico, in vitro and in vivo approaches to analyze various aspects of OmpC's antigenic properties. The conserved region, in addition to secondary and tertiary structures, and linear B cell epitopes, were predicted. A number of results obtained from in silico analyses were validated by experimental studies. OmpC was amplified, cloned and then expressed, with the recombinant protein then being purified. BALB/c mice were immunized by purified denatured OmpC. The titer of antibody was raised. Results of challenges with the pathogen revealed that the immunity is non-protective. Most of the theoretical and experimental results were in consensus. Introduced linear B cell epitopes can be employed for the design of diagnostic kits based on antigen-antibody interactions.

Development of Two Quantitative Real-Time PCR Diagnostic Kits for HPV Isolates from Korea

  • Jeeva, Subbiah;Kim, Nam-Il;Jang, In-Kwon;Choi, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • 제22권10호
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    • pp.1350-1358
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    • 2012
  • Viral pathogens, alongside other pathogens, have major effects on crustacean aquaculture. Hepatopancreatic parvovirus (HPV) is an emerging virus in the shrimp industry and has been detected in shrimp farms worldwide. The HPV genome has greater diversity than other shrimp viruses owing to its wide host range and geographical distribution. Therefore, developing diagnostic tools is essential to detect even small copy numbers from the target region of native HPV isolates. We have developed two easy to use quantitative real-time PCR kits, called Green Star and Dual Star, which contain all of the necessary components for real-time PCR, including HPV primers, using the primers obtained from the sequences of HPV isolates from Korea, and analyzed their specificity, efficiency, and reproducibility. These two kits could detect from 1 to $1{\times}10^9$ copies of cloned HPV DNA. The minimum detection limits obtained from HPV-infected shrimp were $7.74{\times}10^1$ and $9.06{\times}10^1$ copies in the Green Star and Dual Star assay kits, respectively. These kits can be used for rapid, sensitive, and efficient screening for HPV isolates from Korea before the introduction of postlarval stages into culture ponds, thereby decreasing the incidence of early development of the disease.

COVID-19 전후 의료 진단 특허 출원 동향 분석 (Patent Analysis in the Clinical Diagnosis Sector : Before and After COVID-19)

  • 한유진;박선주
    • 대한예방한의학회지
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    • 제26권2호
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    • pp.25-35
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    • 2022
  • Objectives : This study aims to analyze the patents filed in the clinical diagnosis sector where technologies have been actively developed since the advent of the 4th industrial revolution. Methods : The analysis has been conducted in two ways - the period from 2016 to 2021 and the time points before and after COVID-19 - by visualizing based on the word cloud method. Results : Over two thirds of patents has been filed in the A61B sector (71.8%) and cure, sensor, self diagnosis, control, and breakdown have been observed in the period above. During the overall period (2016~2021), 'ultrasound'(7.5%), 'image'(5.1%), 'skin'(4.0%), 'treatment'(3.4%), and 'artificial intelligence(2.5%)' were the frequently patent applications technologies. In addition, 'ultrasound'(6.2%), 'image'(5.5%), 'skin'(4.0%), 'treatment' (3.7%), and 'portable'(1.7%) appeared most frequently before COVID-19 whereas 'ultrasound(5.5%)', 'artificial intelligence(4.2%)', 'diagnostic device'(1.9%), 'dimentia'(1.6%), and 'diagnostic kit'(1.4%) emerged the most after COVID-19. Conclusion : This study is meaningful in that it showed the technological development trend in the digital diagnosis sector and it was found that the Korean medicine field should contribute to this field more actively in the future.

A Rapid Diagnostic Test for Toxoplasmosis using Recombinant Antigenic N-terminal Half of SAG1 Linked with Intrinsically Unstructured Domain of GRA2 Protein

  • Song, Kyoung Ju;Yang, Zhaoshou;Chong, Chom-Kyu;Kim, Jin-Soo;Lee, Kyung Chan;Kim, Tong-Soo;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • 제51권5호
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    • pp.503-510
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    • 2013
  • Toxoplasma gondii is an apicomplexan parasite with a broad host range of most warm-blooded mammals including humans, of which one-thirds of the human population has been infected worldwide which can cause congenital defects, abortion, and neonatal complications. Here, we developed a rapid diagnostic test (RDT) for T. gondii infection. Antigenic N-terminal half of the major surface antigen (SAG1) was linked with intrinsically unstructured domain (IUD) of dense granule protein 2 (GRA2). The recombinant GST-GRA2-SAG1A protein was successfully expressed and purified as 51 kDa of molecular weight. Furthermore, antigenicity and solubility of the rGST-GRA2-SAG1A protein were significantly increased. The overall specificity and sensitivity of GST-GRA2-SAG1A loaded RDT (TgRDT) were estimated as 100% and 97.1% by comparing with ELISA result which uses T. gondii whole cell lysates as the antigen. The TgRDT tested with Uganda people sera for field trial and showed 31.9% of seroprevalence against T. gondii antibody. The TgRDT is proved to be a kit for rapid and easy to use with high accuracy, which would be a suitable serodiagnostic tool for toxoplasmosis.

진단키트 자동 판독을 위한 영상 분석 기법 (Image Analysis Technique for Automatic Recognition of Diagnostic Kit)

  • 정중은;김주연;배혜수;김호준
    • 한국정보처리학회:학술대회논문집
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    • 한국정보처리학회 2013년도 추계학술발표대회
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    • pp.1309-1311
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    • 2013
  • 본 연구에서는 의료 진단키트의 자동판독 시스템에서, 통제되지 않은 조명 환경에서도 정확한 색상 판별을 위한 ROI 영역 추출 기법과 조명 보정 기법을 고찰한다. 3단계로 세분된 ROI 추출 과정은 조명변화에 적응적인 배경영상 정보를 유지하고, 노이즈 제거와 에지 추출 과정을 포함한다. 진단 결과의 정량적 판별에 중요 지표가 되는 색상정보가 조명의 영향의 의해 왜곡되는 것을 보완하기 위하여 표본 추출된 학습데이터로부터 조명 보정 곡선을 생성한다. 20종류의 색상패턴을 대상으로 적용한 실험 결과를 통하여 제안된 이론의 유용성을 고찰한다.