• Title/Summary/Keyword: detection kit

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Effect of Spinal Cord Removal before or after Splitting and Washing on CNST Decontamination of Beef Carcasses

  • Lim, D.G.;Kim, D.H.;Lee, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.11
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    • pp.1770-1776
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    • 2007
  • Beef carcasses were examined to explore the effects of spinal cord removal and washing on central nervous system tissue (CNST) decontamination of the surface during the slaughtering process. A total of 15 carcasses were split by sawing centrally down the vertebral column and left sides of split carcasses were used for analysis. Samples were collected by swabbing the surface from 4 defined parts on the interior and 4 on the exterior of carcasses from the abattoir and analyzed using an ELISA-based test. The results showed that automatic and manual spray washing decreased CNST contamination, especially on the interior ventral parts of carcass surfaces (p<0.01), but did not decrease CNST on the interior dorsal parts. Increasing washing time to 60 s did not affect the reduction of CNST contamination. Samples following spinal cord removal prior to splitting showed lower calculated levels of "risk material" than the stated limit of detection (0.1%) of the ELISA kit on interior and exterior carcass parts (p<0.01). Therefore, spinal cord removal prior to splitting could be a very effective way to minimize CNST contamination of beef carcasses.

Forensic STR Analysis of Mixed Chimerism after Allogeneic Bone Marrow Transplantation

  • Eom, Yong-Bin
    • Biomedical Science Letters
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    • v.16 no.3
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    • pp.193-196
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    • 2010
  • Multiplex PCR-based short tandem repeat (STR) analysis is considered as a good tool for monitoring bone marrow engraftment after sex-mismatched allogeneic transplantation and provides a sensitive and accurate assessment of the contribution of both donor and/or recipient cells in post-transplantation specimens. Forensic STR analysis and quantitative real time PCR are used to determine the proportion of donor versus recipient each contained within the total DNA. The STR markers were co-amplified in a single reaction by using commercial $PowerPlex^{(R)}$ 16 system and $AmpFISTR^{(R)}$ $Identifiler^{(R)}$ / $Yfiler^{(R)}$ PCR amplification kits. Separation of the PCR products and fluorescence detection were performed by ABI $PRIS^{(R)}$ 3100 Genetic Analyzer with capillary electrophoresis. The $GeneMapper^{TM}$ ID software were used for size calling and analysis of STR profiles. Extracted DNA was quantified by the $Quantifiler^{TM}$ Human DNA / Y Human Male DNA Quantification Kit The intent of this study was to analyze the ratio of donor versus recipient cells in the post-transplant peripheral blood, spleen, lung and kidney specimens. Specimens were taken from the traffic accident male victim who had been engrafted from bone marrow female donor. Blood and spleen specimens displayed female donor DNA profile. Kidney specimen showed male recipient DNA profile. Interestingly, lung tissue showed mixed profiles. The findings of this study indicate that the forensic STR analysis using fluorescence labeling PCR combined with capillary electrophoresis is quick and reliable enough to assess the ratio of donor versus recipient cells and to monitor the mixed chimeric patterns.

Early Detection of Epiphytic Anthracnose Inoculum on Phyllosphere of Diospyros kaki var. domestica

  • Lee, Jung-Han;Han, Ki-Soo;Lee, Sun-Cheol;Shim, Chang-Ki;Bae, Dong-Won;Kim, Dong-Kil;Kim, Hee-Kyu
    • The Plant Pathology Journal
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    • v.20 no.4
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    • pp.247-251
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    • 2004
  • We developed a polyclonal antibody (PAh) based- ELISA system to accurately and rapidly monitor inocula on plant surface before onset of anthracnose. Titer of mouse antisera against conidia of Colletotrichum gloeosporioides was determined by using indirect ELISA. It was high enough to be detectable up to ${\times}$ 12,800 dilutions. Absorbance readings exceeded (1.5even at a 10$^{-5}$ dilution. Sensitivity of PAb was precise enough to detect spore concentration as low as 50 conidia/well by indirect ELISA. PAb1 and PAb2 proved to be very sensitive and highly specific to the target pathogen, C. gloeosporioides, apparently discriminating other unrelated pathogens, or epiphytes. Absorbance values for original isolate exceeded 1.0, but no reaction was detected with other isolates, except three other anthracnose fungi: C. gloeosporioides (pepper strain), Glomerella cingulata (apple strain) and C. lagenarium. Our data suggest that PAb1 and PAb2 bind with the protein epitope that partially contains residues of amino acid, arginine, and Iysine. This kit fulfills the require-ments for detecting inoculums before infection and during onset of anthracnose on sweet persimmon.

Application of Epstein-Barr Virus Cell Lines (CCL85 EB-3) in Performing the EBER mRNA In Situ Hybridization as a Positive Control (Epstein-Barr 바이러스 인사이투 보결합 시행시 양성대조표지로서의 버키트 림프종 세포주 (CCL85 EB-3)의 응용)

  • Kim, Sung-Sook;Han, Woon-Sup;Suh, Joo-Young;Huh, Joo-Ryung
    • The Korean Journal of Cytopathology
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    • v.7 no.1
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    • pp.38-43
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    • 1996
  • Epstein-Barr virus(EBV) is associated with a wide spectrum of benign and malignant disorders including leukoplakia, Hodgkln's lymphoma, central nervous system lymphoma, peripheral T cell lymphoma and nasopharyngeal undifferentiated carcinoma. There are several distinctive aspects of biology of the virus that are important in investigation of virus in clinical specimens. The abundant expression of the EBER mRNA transcripts makes possible the sensitive detection of latent expression in EBV-associated tumors. Although there has been a dramatic increased interest in the direct characterization of EBV in clinical specimens, there have been few studios about the effective and reliable positive controls in performing in situ hybridization technique for EBV, especially on paraffin-em bedded tissue. We applied Burkitts lymphoma ceil line as positive control in EBV in situ hydridization using Oncor Kit. The cell block of Burkitt lymphoma cell line(CCL85 EB-3) showed strong and specific positivity for EBER in situ in nuclei of EBV infected cells.

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Curcumin Induces Caspase Mediated Apoptosis in JURKAT Cells by Disrupting the Redox Balance

  • Gopal, Priya Kalyan;Paul, Mausumi;Paul, Santanu
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.93-100
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    • 2014
  • Background: Curcumin has has been reported to exert anti-inflammatory, anti-oxidation and anti-angiogenic activity in various types of cancer. It has also been shown to induce apoptosis in leukemia cells. We aimed to unravel the role of the redox pathway in Curcumin mediated apoptosis with a panel of human leukemic cells. Materials and Methods: In this study in vitro cytotoxicity of Curcumin was measured by MTT assay and apoptotic effects were assessed by annexin V/PI, DAPI staining, cell cycle analysis, measurement of caspase activity and PARP cleavage. Effects of Curcumin on intracellular redox balance were assessed using fluorescent probes like $H_2DCFDA$, JC1 and an ApoGSH Glutathione Detection Kit respectively. Results: Curcumin showed differential anti-proliferative and apoptotic effects on different human leukemic cell lines in contrast to minimal effects on normal cells. Curcumin induced apoptosis was associated with the generation of intracellular ROS, loss of mitochondrial membrane potential, intracellular GSH depletion, caspase activation. Conclusions: As Curcumin induces programmed cell death specifically in leukemic cells it holds a great promise as a future therapeutic agent in the treatment of leukemia.

Pulsed γ-ray emission from magnetar 1E 2259+586

  • Wu, Jason Hung Kit;Hui, Chung Yue;Huang, Regina Hsiu Hui;Kong, Albert Kwok Hing;Cheng, Kwong Sang;Takata, Jumpei;Tam, Pak Hin Thomas;Wu, Eric Man Ho;Liu, Joe
    • Journal of Astronomy and Space Sciences
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    • v.30 no.2
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    • pp.83-85
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    • 2013
  • Anomalous X-ray pulsars (AXPs) are thought to be magnetars which are young isolated neutron stars with extremely strong magnetic fields of > $10^{14}$ Gauss. Their tremendous magnetic fields inferred from the spin parameters provide a huge energy reservoir to power the observed X-ray emission. High-energy emission above 0.3 MeV has never been detected despite intensive search. Here, we present the possible Fermi Large Area Telescope (LAT) detection of ${\gamma}$-ray pulsations above 200 MeV from the AXP, 1E 2259+586, which puts the current theoretical models of ${\gamma}$-ray emission mechanisms of magnetars into challenge. We speculate that the high-energy ${\gamma}$-rays originate from the outer magnetosphere of the magnetar.

Diagnosis of porcine reproductive and respiratory syndrome (PRRS) and its serological survey using the reverse transcription and polymerase chain reaction (RT-PCR) and ELISA (RT-PCR과 ELISA를 이용한 PRRS 진단 및 항체가 조사)

  • Chu Keum-Suk;Han Keu-Sam;Han Jae-Cheol;Song Hee-Jong
    • Korean Journal of Veterinary Service
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    • v.27 no.3
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    • pp.273-280
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    • 2004
  • The studies were performed for the PRRS antigen and antibody detection from breeding farms, artificial insemination(AI) center and growing farms in Jeonbuk province. 1. Specific PRRS primers were successfully amplified ORF6 617bp and ORF7 448 bp on agarose gel. 2. RT-PCR method has been establish by commercial kit and the thermal cycler program consisted of 30 cycles: $95^{\circ}C$ for 30 sec, $45^{\circ}C$ for 30 sec, and $72^{\circ}C$ for 45 sec. 3. The results of PRRS antibody test by ELISA method in AI centers were $6.6\%,\;53.3\%$ and breeding farms $65\%,\;65\%\;and\;38.7\%$, respectively. The serological positive of the antibody in gilt higher than sow. 4. The sero-positive of the PRRS antibody showed average $21\%$ in domestic farms, $56.2\%$ in breeding farms, and $29.9\%$ in AI center.

Survey on the Distrributions of Swine Toxoplasma Antibodies by Latex Agglutination Test in Gyeongnam Central Area (경남 중부지역에서의 Latex응집반응을 이용한 돼지 톡스플라즈마병 항체분포 조사)

  • 이병훈;황보훈;변유성;이순선;김차용;서명득
    • Korean Journal of Veterinary Service
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    • v.15 no.2
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    • pp.174-183
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    • 1992
  • This study was conducted to determine the serum antibodies against toxoplasma in swine from breeding-pig farm, pig farm and abattoir by latex agglutination(LA) test. LA test was carried out with commercial Toxo-MT kit (Eiken chemical co.). The results obtained were summerized as follows : 1. The cut-off titer of positive and negative reactions by Toxo-MT antigen used in this experiment was determined as the serum dilution of 1 ; 32. 2. positive rates of toxoplasma antibodies in 823 swine sera were 17.0%(140 cases) by LA test. 3. The toxoplasma antibody detection rates against 194 swine sera in breeding-pig farm, 273 swine sera in pig farm and 356 swine in abattoir were 46.9%(91 cases), 8.4%(23 cases) and 7.3% (26 cases) , respectively. 4. In LA test serum antibody titers in 823 test sera were shown as 51 cases (36.4%) in 1 : 32, 40(28.6%) in 1;64, 17(12.1%) in 1:128, 14(10.0%) in 1:256, 10(7.1%) in 1:512, 5(3.6%) in 1:1,024, and 3(2.1%) in 1 : 2,048. 5. Positive rates of toxoplasma antibodies in swine sera from each breeding-pig farm were 20.0∼61.9%.

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Development of PCR-microplate Hybridization Assay for Detection of Mycobacterium tuberculosis

  • Lee, In-Soo;Cho, Een-Jin;Cho, Sang-Nae;Kim, Tae-Ue;Lee, Hye-Young
    • Biomedical Science Letters
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    • v.15 no.4
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    • pp.295-300
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    • 2009
  • Tuberculosis caused by Mycobacterium tuberculosis (MTB) still remains to be the most dreadful infectious disease affecting almost every country. In the present study, we developed a simple and rapid but accurate and sensitive assay method for detecting MTB using microplate hybridization assay. For this, a selective region of the rpoB gene was used to design PCR primers, and MTB and Mycobacterium genus-specific probe molecules. The specificity of the assay was confirmed using fifteen different mycobacterial reference strains and twelve different non-mycobacterial reference strains, and the sensitivity was determined to be 100 fg using genomic DNA of MTB reference strain, H37Rv. Subsequently, a total of 62 sputum samples with diverse smear scores and culture positive results were used to evaluate the kit performance. In brief, the specificity and the sensitivity of the assay were 100% and 98.4%, respectively.

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UV-HPLC Determination of Carbowyl Group Using 2-Bromoacetyltriphenylene as a Pre-labeling Reagent - The isolative determination of prostaglandin $E_2$ and $F_2{\alpha}$ by HPLC (2-Bromoacetyltriphenylene 유도체화제를 이용한 카르복실기 함유성분의 분석법 (I) - 프로스타글란딘 $E_2$$F_2{\alpha}$ 혼합물의 HPLC에 의한 분리정량)

  • 이왕규;정해수;김박광
    • YAKHAK HOEJI
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    • v.30 no.6
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    • pp.311-316
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    • 1986
  • A new UV labeling reagent was developed and used in HPLC for the determination of prostaglandin $E_2$ which have weak UV light-absorbing property. This reagent, 2-bromoacetyltriphenylene, was synthesized by the bromination of 2-acetyltriphenylene which was obtained from triphenylene by Friedel-Crafts reaction. The wave length maximum (${\lambda}_{max}^{CH_3CN}$ of this reagent was 268nm. Prostaglandin E$_2$ was extracted from prostaglandin E$_2$-$\beta$-cyclodextrin using a Sep-pak $C_{18}$ cartridge. The prostaglandin E$_2$ was labeled with 2-bromoacetyl-triphenylene in aectonitrite using 18-crown-6-ether as catalyst. Derivatized prostaglandins were separated on a reversed-phase column (Radial-pak) $\mu$-Bondapak $C_{18}$ using acetonitrile: water=60:40 as mobile phase. The effluent was monitored by UV detector at 254nm filter kit. Linearity of calibration curve was obtained between 30ng and 140ng, and the lower limit of detection was 5ng.

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