• 제목/요약/키워드: assay

검색결과 13,998건 처리시간 0.031초

합성기질 및 응고시간을 이용한 혈액응고 제 8인자 역가측정법 (Potency Assay of Factor Ⅷ : C Concentrates using the Chromogenic and Clotting Assay)

  • 강혜나;김순남;허숙진;홍성화
    • 약학회지
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    • 제45권5호
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    • pp.476-483
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    • 2001
  • The clotting assay was replaced by the chromogenic substrate assay which is recommended by the European Pharmacopoeia (EP) and the Scientific and Standardization Committee of the International Society on Thrombosis and Haemostasis based on the reliability convenience and simplicity of the chromogenic assay, A correlation study was carried out with a one-stage factor Ⅷ : C clotting assay and the performance of the chromogenic assay was evaluated using two test kits that fulfilled the requirements of EP for factor Ⅷ concentrates test. Although chromogenic assay has partly differences in measurement principle and standardization, this assay has a high correlation with clotting assay in various types of factor Ⅷ concentrates and factor Ⅷ standard. We conclude that the chromogenic assay for factor Ⅷ : C concentrates correlates well with the clotting assay and shows good analytical performance.

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수두바이러스의 정량에 있어서 Solid-overlay Standard Plaque Assay와 Immunofocus Assay의 효용성 비교 연구 (Comparative Study of the Standard Plaque Assay with Solid-overlay and Immunofocus Assay for Varicella-zoster Virus Titration)

  • 이화경;정용석
    • 대한바이러스학회지
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    • 제30권1호
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    • pp.61-70
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    • 2000
  • Standard plaque assay using agarose-overlay has long been used for titration of many infectious virus particle. Plaque assay for the titration of varicella-zoster virus and its live vaccine requires three intermittent agarose overlay to visualize plaques. Overall procedure of the assay takes at least nine days from virus inoculation and microbe contamination including fungi is frequently accompanied during incubation period. We studied whether an immunofocus assay in conjunction with peroxidase-mediated immunohistochemical reaction may replace the standard plaque assay for the virus titration by comparing the two methods. A linear relationship was observed between number of foci and virus dilution. The number of foci in a given dilution of virus appeared a little higher than counted plaques formed in standard plaque assay. Independent titration results obtained from two assay methods for a given dilution of virus demonstrated a strong correlation ($r^2=0.99$). Foci of virus infected cells as revealed by the enzyme reaction could be counted either 4 days post-infection (p.i.) under low magnification (40X) microscopy, or 6 days p.i. by naked eye observation. Larger size of cell cuture plate, virus adsorption at $35^{\circ}C$, and 10% FBS in diluent appeared to be better conditions for the assay. Immunofocus assay will be an effective and dependable titration method for varicella-zoster virus and its live vaccine in place of the standard plaque assay in respect to accuracy, costs, and experimental convenience.

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Hollow Fiber 검색법과 Xenographic Animal Assay를 이용한 생약재의 암세포 저해활성 비교 (Comparative Activity of Medicinal Herbs Between Hollow Fiber Assay and Xenographic Animal Assay)

  • 조좌형;윤원호;이경호
    • 생약학회지
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    • 제34권4호통권135호
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    • pp.288-292
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    • 2003
  • We compared the antitumor activity between hollow fiber assay and xenographic animal assay on thirty herbal plants. It is evaluated that the antitumor activity of above 30% is regarded as 'positive response', and its of below 30% is regarded as 'negative response'. The two herbal plants extracts (Ulmus davidiana Hedyotis diffusa) among thirty herbal plants show to be positive in xenographic animal assay and they were also correctly identified as positive by the hollow fiber assay. The correlation of the hollow fiber assay data with xenographic animal assay would suggest that hollow fiber assay presents a potentially unique tool to develop the herbal medicine for cancer.

랫드에서 Uterotrophic assay 및 Hershberger assay를 이용한 칡의 에스트로겐/항안드로겐 영향 평가 (The Evaluation of Estrogenic/Antiandrogenic Activity of Puerariae Radix in Immature Rats Using Uterotrophic Assay and Hershberger Assay)

  • 곽승준;김순선;이규식;손경희;김희연;강길진;최요우;박철훈;박귀례
    • Toxicological Research
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    • 제18권4호
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    • pp.393-396
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    • 2002
  • This study was carried out to evaluate the ostrogenic/antiandrogenic activity of Puerariae Radix in Sprague-Dawley rats. It has known that diverse phytoestrogen were included in some Puerariae Radix, especially in Pueraria mirifica. The Uterotrophic assay and Hershberger assay were performed to evaluate the ostogenic/antiandrogenic activity of various Puerariae Radix (Pueraria thunbergiana, Pueraria mirifica and Butea superba). In Uterotrophic assay, the extracts of Puerariae Radix were administered subcutaneously to immature female SD rats from 19 to 21 days of age. The wet uterus and vaginal weighs significantly increased in the group only treated with extracts of Pueraria mirifica. But, in Hersh-berger assay, all extracts of Puerariae Radix did not show any effects in the castrated rats. These results suggest that Pueraria mirifica has not undrogenic/antiandrogenic effect but potent estrogenic effect. It is possible that components of Pueraria mirifica may act as endocrine disruptor in human body.

미생물 배양액으로부터 항암제의 예비선별을 위한 cccDNA Breakage 활성검정과 Assay Sulforhodamine B 활성검정의 이용 (Use of cccDNA Breakage Assay and Sulforhodamine B Assay for the Prescreening of Antitumor Agents from Microbial Sources)

  • 이상한;이동선;김종국;홍순덕
    • 생명과학회지
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    • 제8권1호
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    • pp.67-71
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    • 1998
  • 다량의 시료를 빠르게, 적은 격비로, 간다히 검색할수 있는 예비선별법을 찾던중, ccDNA와 sulforhodamine B를 각각 사용한 미생물배양액내의 신규항암후보물질의 탐색을 시도하였다. cccDNA법으로는 3.3%가 positive 반응을 나태내었으며, SRB assat에서는 A549와 SK-OV-3에 활성을 나타내는 4개의 발효여액을 확인한수 있었다.

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Validation of Kinetic Method for the PKA Assay in Plasma-Derived Products

  • Shin, In-Soo;Hong, Choong-Man;Koh, Hyun-Chul;Hong, Seung-Hwa
    • Biomolecules & Therapeutics
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    • 제13권1호
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    • pp.59-63
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    • 2005
  • A kinetic assay was carried out in order to compare the ability of detection for prekallikrein activator(PKA) in plasma-derived products with that of an endpoint assay and a commercial method. Using these methods, 9 human albumin preparations were assayed and compared to each other. The coefficient of variation between the Kinetic assay and the end point assay was found within 6.6% and this result showed that two methods were highly correlative and the end point assay could act as a replacement of the kinetic assay. Another important goal of this study was to investigate the reproducibility among laboratories on the kinetic assay. A collaborative study was performed to validate the kinetic method with intra and inter assays. The coefficient of variation for the intra assay of each laboratory was less than 4% and that for between individuals in the inter assay was 4.1%. These results revealed that the kinetic assay showed good reproducibility. The contents of PKA in plasma-derived products were also determined by the kinetic assay. As a result, it was found that trace amounts of PKA were present in 32 human immunoglobulin preparations, however the average concentration of PKA in 171 albumin preparations was 5.8 IU/mL.

핵의학 체외 검사에서 자동분주기를 이용한 Random Assay 가능성평가 (Comparison of Batch Assay and Random Assay Using Automatic Dispenser in Radioimmunoassay)

  • 문승환;이호영;신선영;민경선;이현주;장수진;강지연;이동수;정준기;이명철
    • Nuclear Medicine and Molecular Imaging
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    • 제43권4호
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    • pp.323-329
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    • 2009
  • 목적: 방사면역측정법 및 면역방사계수법은 매 실험마다 표준용액을 넣고 표준곡선에 대비하여 결과를 산출하는 batch assay가 일반화되어 있다. 비용과 시간에 대한 효율성을 증가 시키기 위해서는 선행 표준 곡선을 이용하여 검사를 진행하는 random assay의 적용이 요구된다. 방사면역측정법 및 면역방사계수법에서 자동분주기를 이용한 random assay적용 가능성을 평가 하기 위해 본 연구를 수행하였다. 대상 및 방법: Triiodothyronine (T3), free thyroxine (fT4), Prostate specific antigen (PSA), Carcinoembryonic antigen (CEA) 4가지 항목을 대상으로 하였으며 각 검사별 20명의 환자 검체를 이용하였다. 각 검사 항목별로 표준곡선을 얻기 위해 3시간 간격으로 4회 측정한 표준용액의 분당계수값(counter per minute, cpm)을 측정하였다. 이를 Friedman test를 이용하여 비교하여 검사시 마다 얻는 표준 곡선간의 차이를 평가하였다. 각 검사별 저, 중, 고농도의 대조 용액을 이용하여 5회 측정하고 이를 Friedman test를 이용하여 비교하여 측정 내 정밀도를 평가하였다. 각 검사별 저, 중, 고농도의 대조용액을 batch assay와 자동분주기를 이용한 random assay로 3시간 간격으로 각각 4회 측정하였다. 측정값을 바탕으로 평균, 표준 편차, 변이계수를 구하고 Wilcoxon test를 이용하여 비교하여 assay 방법에 따른 차이를 비교하였다. 두 assay간에 일치도(agreement)를 평가하기 위해 두 assay의 측정값 사이에 급내상관계수(Intraclass correlation coefficient, ICC)와 상관계수를 계산하였다. 결과: 모든 검사항목에서 표준 용액의 분당계수값은 측정 간에 통계적으로 유의한 차이가 없었다. T3, fT4, PSA, CEA 검사항목마다 측정한 저, 중, 고농도별 측정 내 변이계수는 T3(3.6%, 2.6%, 0.9%), fT4(4.5%, 1.1%. 1.3%), PSA(3.9%, 3.1%, 1.2%), CEA(5.0%, 4.8%, 7.6%)였다. 검사 항목별 측정방법에 따른 대조 용액을 이용한 표준 곡선의 측정 간 정밀도 평가에서 두 검사 방법간에 유의한 차이가 없었다. 각 검사항목별로 20명의 환자 검체에서 측정한 측정 간 변이계수(inter-assay precision)의 평균과 표준편차 값은 batch assay로 시행했을 때 각각 3.2$\pm$1.7%, 3.9$\pm$2.1%. 7.1$\pm$6.2%, 11.2$\pm$7.2%였고 random assay로 측정하였을 때는 2.7$\pm$1.7%, 4.8$\pm$3.1%, 3.6$\pm$4.8%, 7.4$\pm$6.2였다. Batch assay와 random assay는 서로 높은 일치도를 보였다. 측정한 급내상관계수(Intraclass correlation coefficient)는 T3, fT4, PSA, CEA 각각 0.9968, 0.9973. 0.9996, 0.9901였다. Batch assay로 측정한 값과 자등분주기를 이용한 random assay로 측정한 값 사이에 상관 계수(R$^2$)는 T3, fT4, PSA, CEA 각각 0.9924, 0.9974, 0.9994, 0.9989(p<0.005)로 모두 강한 상관관계를 보였다. 결론: T3, fT4, PSA, CEA 4가지 항목의 방사면역측정 법 (Radioimmunoassay)에서 기존의 batch assay와 자동분주기를 이용한 random assay간에 일치도와 상관성이 높았다. 결론적으로 자동분주기를 이용한 random assay를 방사면역측정법 및 면역방사계수법에 적절히 이용할 수 있을 것으로 생각된다.

Sulforhodamine B Assay to Determine Cytotoxicity of Vibrio vulnificus Against Human Intestinal Cells

  • Lee, Byung-Cheol;Choi, Sang-Ho;Kim, Tae-Sung
    • Journal of Microbiology and Biotechnology
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    • 제14권2호
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    • pp.350-355
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    • 2004
  • Sulforhodamine B (SRB) assay is a rapid, sensitive, and inexpensive method for measuring cell proliferation and chemosensitivity. However, the lactate dehydrogenase (LDH) release assay is generally used to measure cytototoxicity of infectious microorganisms against host cells. In this study, we investigated the possibility of applying the SRB assay to determine cytotoxicity for infectious microorganisms, and compared the results with those obtained by the LDH release assay. We used Vibrio vulnificus as a model of infectious microorganisms. The SRB assay showed that V vulnificus strongly induced cytotoxic activity against human intestinal cells, Caco-2 and INT-407 cells. The degree of cytotoxicity closely correlated with infection time and number ratios of V. vulnificus to intestinal cells (MOI, multiplicity of infection). Furthermore, cytotoxicity values obtained by SRB assay correlated well with results obtained by the LDH release assay, and both assays gave a linear response with respect to MOI Heat-inactivation of V. vulnificus for 35 min at $60^{\circ}C$ did not induce cytotoxic activity, indicating that viability of V. vulnificus is crucial for cytotoxic activity against intestinal cells. Although both assays are suitable as cytotoxicity endpoints, the SRB assay is recommended for measuring cytotoxicity of infectious microorganisms against host cells because of its significantly lower cost and more stable endpoint than the LDH release assay.

Quantitative Detection of Residual E. coli Host Cell DNA by Real-Time PCR

  • Lee, Dong-Hyuck;Bae, Jung-Eun;Lee, Jung-Hee;Shin, Jeong-Sup;Kim, In-Seop
    • Journal of Microbiology and Biotechnology
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    • 제20권10호
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    • pp.1463-1470
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    • 2010
  • E. coli has long been widely used as a host system for the manufacture of recombinant proteins intended for human therapeutic use. When considering the impurities to be eliminated during the downstream process, residual host cell DNA is a major safety concern. The presence of residual E. coli host cell DNA in the final products is typically determined using a conventional slot blot hybridization assay or total DNA Threshold assay. However, both the former and latter methods are time consuming, expensive, and relatively insensitive. This study thus attempted to develop a more sensitive real-time PCR assay for the specific detection of residual E. coli DNA. This novel method was then compared with the slot blot hybridization assay and total DNA Threshold assay in order to determine its effectiveness and overall capabilities. The novel approach involved the selection of a specific primer pair for amplification of the E. coli 16S rRNA gene in an effort to improve sensitivity, whereas the E. coli host cell DNA quantification took place through the use of SYBR Green I. The detection limit of the real-time PCR assay, under these optimized conditions, was calculated to be 0.042 pg genomic DNA, which was much higher than those of both the slot blot hybridization assay and total DNA Threshold assay, where the detection limits were 2.42 and 3.73 pg genomic DNA, respectively. Hence, the real-time PCR assay can be said to be more reproducible, more accurate, and more precise than either the slot blot hybridization assay or total DNA Threshold assay. The real-time PCR assay may thus be a promising new tool for the quantitative detection and clearance validation of residual E. coli host cell DNA during the manufacturingprocess for recombinant therapeutics.

Head-to-Head Comparison between Xpert MTB/RIF Assay and Real-Time Polymerase Chain Reaction Assay Using Bronchial Washing Specimens for Tuberculosis Diagnosis

  • Son, Eunjeong;Jang, Jinook;Kim, Taehwa;Jang, Jin Ho;Chung, Jae Heun;Seol, Hee Yun;Yeo, Hye Ju;Yoon, Seong Hoon;Lee, Seung Eun;Cho, Woo Hyun;Kim, Yun Seong;Jeon, Doosoo
    • Tuberculosis and Respiratory Diseases
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    • 제85권1호
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    • pp.89-95
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    • 2022
  • Background: With the introduction of Xpert MTB/RIF assay (Xpert), its incorporation into tuberculosis (TB) diagnostic algorithm has become an important issue. The aim of this study was to evaluate the performance of the Xpert assay in comparison with a commercial polymerase chain reaction (PCR) assay. Methods: Medical records of patients having results of both Xpert and AdvanSure TB/NTM real-time PCR (AdvanSure) assays using the same bronchial washing specimens were retrospectively reviewed. Results: Of the 1,297 patients included in this study, 205 (15.8%) were diagnosed with pulmonary TB. Using mycobacterial culture as the reference method, sensitivity of the Xpert assay using smear-positive specimens was 97.5%, which was comparable to that of the AdvanSure assay (96.3%, p=0.193). However, the sensitivity of the Xpert assay using smear-negative specimens was 70.6%, which was significantly higher than that of the AdvanSure assay (52.9%, p=0.018). Usng phenotypic drug susceptibility testing as the reference method, sensitivity and specificity for detecting rifampicin resistance were 100% and 99.1%, respectively. Moreover, a median turnaround time of the Xpert assay was 1 day, which was significantly shorter than 3 days of the AdvanSure assay (p<0.001). Conclusion: In comparison with the AdvanSure assay, the Xpert assay had a higher sensitivity using smear-negative specimens, a shorter turnaround time, and could reliably predict rifampin resistance. Therefore, the Xpert assay might be preferentially recommended over TB-PCR in Korean TB diagnostic algorithm.