• Title/Summary/Keyword: Kit

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Fluorogenic and Chromogenic Assay for Rapid Detection of Escherichia coli and Total Coliform Bacteria (효소발색법을 이용한 대장균 및 총대장균군 신속 검사)

  • Lee, Keun-Heon;Kim, Hun-Soo;Kim, Byong-Ryol;Lee, Seung-Hui;In, Chi-Kyung;Park, Kyeong-Ryang
    • Journal of Korean Society on Water Environment
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    • v.25 no.3
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    • pp.363-369
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    • 2009
  • We developed the Eco medium for Escherichia coli and total coliforms, which was modified by Violet Red Bile (VRB) medium, and derived the standard curve of exponential phase at $OD_{410}$ by using type strains such as E. coli ATCC11303, Enterobacter cloacae KCTC2361, Klebsiella pneumoniae KCTC2241, and Citrobacter freundii KCTC2359. Also, we used total 93 samples of spring and stream water to compare the detection ability of total coliforms between the method using Eco medium and such as most probable number (MPN), and plate count methods. As a result, the qualitative analysis of E. coli and total coliforms using Eco medium contained ortho-nitrophenyl-$\beta$-galactoside (ONPG) and 4-methylumbelliferyl-$\beta$-D-glucuronide (MUG) was same as those of Korean standard methods (Colilert kit). And the colony forming unit (CFU) detected in Eco medium was similar to those of result from MPN and plate count methods. Moreover, the agreement, sensitivity, and specificity of the developed kit was more than 97.5% in comparison with Colilert kit for 350 samples. Thus, the Eco medium can be used both qualitative and quantitative analysis of E. coli and total coliforms.

Comparison of the non-invasive diagnostic methods, stool antigen test and PCR assay, for Helicobacter felis detection in dogs

  • Hong, Sunhwa;Lee, Hak-Yong;Kim, Tae-Wan;Kim, Okjin
    • Korean Journal of Veterinary Service
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    • v.38 no.1
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    • pp.37-42
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    • 2015
  • The aim of the present study was to compare the non-invasive methods for the diagnosis of H. felis with HpSA kit-based detection method and H. felis-specific PCR assay with dog's stool samples without sacrifice. Male Beagle dogs (n=6) were infected with H. felis ATCC 49179 ($1.0{\times}10^9CFU/dog$) by intra-gastric inoculation two times at 3-day intervals, and the stool specimens of dogs were collected 1, 3, 5, 7, 14, 21 days after infection to submit to HpSA test and H. felis-specific PCR. As the results, the sensitivity of the HpSA and the PCR analysis was 50.0%, 83.3% respectively. Although HpSA test is less sensitive, it could be used for rapid, cheap and easy screening assay for H. felis infection in dog and cats. We suggest that the H. pylori stool antigen kit, HpSA, is useful and effective for monitoring H. felis infection. If HpSA test would be made with H. felis antibodies in the future, its sensitivity could be increased. Also, PCR assay could be successfully used to detect the H. felis in stools. Applying the H. pylori stool antigen kit and PCR assay may be the recommended non-invasive strategy to identify H. felis in dog and cats.

Effect of Gleevec on Head and Neck Squamous Cell Carcinoma (두경부편평세포암종에서 Gleevec의 효과)

  • Chu Hyung-Ro;Weisman Robert A.
    • Korean Journal of Head & Neck Oncology
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    • v.21 no.2
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    • pp.158-164
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    • 2005
  • Purpose: The serine/threonine kinase Akt was described to inhibit apoptosis in cancer. This study was to examine the effect of Gleevec on head and neck squamous cell carcinoma(HNSCC) through the mechanism of Akt. Experimental Design: Gleevec was introduced into the HNSCC cell lines UMSCC10B, HN12 and HN30 in a range of concentrations. Cell viability was assessed by clonogenic survival analysis. Targets of Gleevec(PDGFR, c-Kit, and c-Abl) were evaluated by Western blot. HNSCC tissue samples were stained for PDGFR, c-Kit and phosphorylated Akt. Akt phosphorylation following Gleevec treatment was assessed using Western blot. Akt siRNA was used to as the positive control. Results: Colony forming efficiency decreased with an increase in concentration of Gleevec. Expressions of PDGFR, c-Kit, and c-Abl were observed in HNSCC cells. Immunohistochemistry confirmed high expression of PDGFR, c-Kit, and p-Akt in human HNSCC tissues. Akt kinase activity was significantly inhibited with increasing concentration of Gleevec in HNSCC cells, and near complete dephosphorylation of Akt was observed at $6{\mu}M$ of Gleevec in the UMSCC10B and HN30 cell lines. Conclusions: Gleevec at clinically comparable concentrations caused a dose dependant decrease in HNSCC survival. The decreased cell survival was related to the inhibition of Akt kinase activity and dephosphorylation of Akt. Akt signaling pathway may be a relevant target for Gleevec in treating HNSCC.

Characterization of Titanium Diboride Composite Bipolar Plate for Polymer Electrolyte Membrane Fuel Cell (전해질 연료전지용 복합분리판의 특성에 미치는 TiB2 첨가효과)

  • Park, Jong-Moon;Sohn, Je-Ha;Park, Yong-Il;Lee, Dong-Gu;Oh, Myung-Hoon
    • Journal of the Korean Society for Heat Treatment
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    • v.27 no.4
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    • pp.169-174
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    • 2014
  • The effect of varying amounts of graphite and $TiB_2$ on the electrical conductivity of composite bipolar plates was systematically studied. In this study, Titanium diboride ($TiB_2$) which has a high electrical conductivity, was selected as a filler and a additive material instead of conventional graphite. For proper distribution of the filler and matrix materials, ball milling using alumina balls was conducted for 1h, and then the hot press method was applied for the preparation of composite samples. The results showed a rapid increase in the electrical conductivity of composite bipolar plates at the critical filler content. However, $TiB_2$ and graphite composite bipolar plates showed similar increases in the electrical conductivity even though $TiB_2$ has a higher electrical conductivity than graphite. In addition, it was also found that a small addition of $TiB_2$ to graphite filler could be very effective for increasing the electrical conductivity and flexural strength of the composite bipolar plate.

DNA Extraction from Protozoan Oocysts/Cysts in Feces for Diagnostic PCR

  • Hawash, Yousry
    • Parasites, Hosts and Diseases
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    • v.52 no.3
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    • pp.263-271
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    • 2014
  • PCR detection of intestinal protozoa is often restrained by a poor DNA recovery or by inhibitors present in feces. The need for an extraction protocol that can overcome these obstacles is therefore clear. $QIAamp^{(R)}$ DNA Stool Mini Kit (Qiagen) was evaluated for its ability to recover DNA from oocysts/cysts directly from feces. Twenty-five Giardia-positive, 15 Cryptosporidium-positive, 15 Entamoeba histolytica-positive, and 45 protozoa-free samples were processed as control by microscopy and immunoassay tests. DNA extracts were amplified using 3 sets of published primers. Following the manufacturer's protocol, the kit showed sensitivity and specificity of 100% towards Giardia and Entamoeba. However, for Cryptosporidium, the sensitivity and specificity were 60% (9/15) and 100%, respectively. A series of optimization experiments involving various steps of the kit's protocol were conducted using Cryptosporidium-positive samples. The best DNA recoveries were gained by raising the lysis temperature to the boiling point for 10 min and the incubation time of the InhibitEX tablet to 5 min. Also, using a pre-cooled ethanol for nucleic acid precipitation and small elution volume ($50-100{\mu}l$) were valuable. The sensitivity of the amended protocol to Cryptosporidium was raised to 100%. Cryptosporidium DNA was successfully amplified by either the first or the second primer set. When applied on parasite-free feces spiked with variable oocysts/cysts counts, ${\approx}2$ oocysts/cysts were theoretically enough for detection by PCR. To conclude, the Qiagen kit with the amended protocol was proved to be suitable for protozoan DNA extraction directly from feces and support PCR diagnosis.

An Introduction of Urbani School Health Kit Developed by World Health Organization (세계보건기구의 Urbani School Health Kit 소개)

  • Nam, Eun-Woo;Chang, Chang-Gok;Park, Soon-Woo;Bonito, Sheila;Kim, Tae-Ho;Shin, Hai-Ri
    • The Journal of Korean Society for School & Community Health Education
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    • v.12 no.1
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    • pp.117-129
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    • 2011
  • Objectives: The purpose of this study was to introduce of the Urbani School Health Kit (USHK) and to investigate the adaptability to Korean situation. Methods: The authors analyzed the contents of USHK developed by the health promotion team at the Western Pacific Regional Office of World Health Organization (WHO WPRO) in collaboration with health promotion experts at University of the Philippine Open University, and by observing health classes using the USHK in Angono elementary school in Manila, Philippines. Results: The following are the characteristics of USHK: 1) The USHK was composed of a teacher's guide and six books targeted to two groups of children: ages 5~6 years and 10~12, and an integrated package containing materials that can be used to support health education and health promotion activities in elementary schools. 2) The USHK could be integrated in the curriculum to reduce the burden of teacher's class preparation time and help teachers conveying clear and accurate health messages in their classes. 3) Several evaluation tools such as pre-test post-test quizzes, self-evaluation tools, observer checklists, and evaluation interview protocols were developed to monitor and evaluate whether USHK was useful, helpful, and appropriate. Conclusions: We found that USHK was a practical tool for supporting health promotion in elementary schools and could be applicable to health promoting schools in Korea if it were modified to address Korean school health problems.

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Detection of residual antibiotics by TLC and EEC-4 plate method in slaughtered pigs (도축돈에서 TLC와 EEC-4 plate법을 이용한 항생물질 잔류조사)

  • 권오성;김순태;김영욱;손재권
    • Korean Journal of Veterinary Service
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    • v.20 no.3
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    • pp.313-321
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    • 1997
  • The antibiotic residues of the urine, the liver, the lung, the kidney and the spleen in slaughtered pigs at Kyongbuk province were detected by TLC(505 kit) and EEC-4 plate method. 1. The positive rate of residual sulfamethazine which was detected by 505 kit in the urine (n=200) was 0.0%. 2. The positive rate of residual sulfamethazine which was detected by EEC-4 plate in the urine (n=126), the liver(n=98), the kidney(n=72), the spleen (n=68) and the lung(n=48) were 63%, 49%, 36%, 34% and 24%, respectively. 3. By EEC-4 plate method, the positive detection rates of the urine were 53.0% in BS(pH 6.0), 29.0% in BS(pH 7.2), 11.5% in BS(pH 8.0) and 13.0% in ML(pH 8.0) medium, that of the liver 41.5% in BS(pH 6.0), 22.0% in BS(pH 7.2), 6.5% in BS(pH 8.0) and 5.0%, in ML (pH 8.0) medium, that of the lung 21.0% in BS(pH 6.0), 9.5%, in BS(pH 7.2) and 8.5% in BS(pH 8.0) medium, and that of the kidney 31.5% in BS(pH 6.0), 14.5% in BS(pH 7.2), 20.0% in BS(pH 8.0) and 3.0% in ML(pH 8.0) medium. In the spleen, only in BS(pH 6.0) medium the positive rate was detected as 33.5 %. 4. The positive rates of samples which shown TLC-positive detected by EEC-4 plate method were 53.9% in no band, 77.8% in one band, 80.9% in two bands, 66.7% in three bands, respectively. In conclusion, the EEC-4 plate method could be applied for the detection of residual antibiotics in samples which shown as out of standard Rf values by TLC-method (SOS kit).

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Neurogenic potentials of human amniotic fluid-derived stem cells according to expression levels of stem cell markers and ingredients of induction medium

  • Lim, Eun Hye;Cho, Jung Ah;Park, Ho;Song, Tae Jong;Kim, Woo Young;Kim, Kye Hyun;Lee, Kyo Won
    • Journal of Genetic Medicine
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    • v.12 no.1
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    • pp.31-37
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    • 2015
  • Purpose: We investigated the neurogenic potentials of amniotic fluid-derived stem cells (AFSCs) according to the expression levels of stem cell markers and ingredients in the neural induction media. Materials and Methods: Four samples of AFSCs with different levels of Oct-4 and c-kit expression were differentiated neurally, using three kinds of induction media containing retinoic acid (RA) and/or a mixture of 3-isobutyl-1-methylxanthine/indomethacin/insulin (neuromix), and examined by immunofluorescence and reverse transcription-polymerase chain reaction (RT-PCR) for their expression of neurospecific markers. Results: The cells in neuromix-containing media displayed small nuclei and long processes that were characteristic of neural cells. RT-PCR analysis revealed that the number of neural markers showing upregulation was greater in cells cultured in the neuromix-containing media than in those cultured in RA-only medium. Neurospecific gene expression was also higher in Oct-4 and c-kit double-positive cells than in c-kit-low or -negative cells. Conclusion: The stem cell marker c-kit (rather than Oct-4) and the ingredient neuromix (rather than RA) exert greater effects on neurogenesis of AFSCs.

Development of Rapid Diagnostic Kit for Identification of Hanwoo (Korean Native Cattle) Brand Meat by Detecting BIO-TAG

  • Baek, Kyung Hoon;Park, Sung Kwon;Lee, Myung Hoon;Kim, Sung Il;Cho, Soo Hyun;Choi, Chang Bon
    • Food Science of Animal Resources
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    • v.34 no.3
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    • pp.339-345
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    • 2014
  • This study was performed to develop a rapid immuno-assay kit, by using a specific antigen to detect Hanwoo brand meat. We selected a synthetic antigen specific to our target antibody, named BIO-TAG (Tyr-D-Ala-Phe), by utilizing a computer-based analysis and literature review. BIO-TAG tagged with adjuvant was subcutaneously injected in sheep and Hanwoo. The serum and meat juice of the immunized or non-immunized animal were then analyzed, to measure the titer of antibody by ELISA and Western blot. The amount of antibodies against the BIO-TAG increased (p<0.05) in serum by vaccination. Furthermore, meat juice from the immunized Hanwoo showed greater (p<0.05) antibody titer, compared with those from non-immunized groups. To optimze the dilution factor, we performed dot-ELISA, with various combination levels of BIO-TAG. Results from dot-ELISA showed that 2 mg/mL BIO-TAG was sufficient to distinguish the immunized meat from non-immunized groups. These results support our hypothesis that simple immunization of Hanwoo generates a sufficient amount of antibodies to be detectable in the meat juice by means of the immune-assay. Therefore, specific Hanwoo brand meat can be more precisely identified by our rapid diagnostic kit. This technology can deter possible fraud of counterfeit meat brands in the Korean domestic market with ease and rapidity; and offers a new tool that guarantees consumers high quality Hanwoo brand beef.

The Development of a Taste Kit for Education and Research into Sensory Characteristics (아동 미각교육을 위한 쌀 Kit 개발 및 이를 활용한 미각 특성조사)

  • Kim, Mi-Hye;Chung, Hae-Kyung
    • Journal of the Korean Society of Food Culture
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    • v.28 no.6
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    • pp.585-593
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    • 2013
  • This study was designed to measure taste sensitivity and the five basic senses by an educational classification instrument. The instrument was a rice kit that could use samples in a dry powder form or oil extract after long-term storage To test for taste, sucrose, salt, citric acid, and quinine sulfate were made at different concentrations and taste sensitivity was measured on a scale from level 1 to level 5. To obtain baseline data, an inspection tool for the five senses was used and randomly applied on 101 schoolchildren in the third and fourth grade in the city of Cheonan in Korea. The inspection tool was composed of 17 questions; 5 questions regarding visual characteristics and three questions each for characteristics regarding taste, hearing, smell, and touch. The average age of the schoolchildren was 9.5 years old and there were 49 third grade students (9 years of age), and 52 fourth grade students (ten years of age). There were slightly more male students than female students, 56 (55.4%) compared to 45 (44.6%), respectively. The average height of female students was higher than that of males, but the average BMI (body mass index) of the male students was slightly higher than that of female students (18.28 compared to 18.09, respectively). Female students were slightly more sensitive to salty tastes than male students (2.8 compared to 2.5, respectively). In the score distribution for each sense, touch sense was the highest at 7.59, sight sense was 7.49, hearing sense was 5.43, smell sense was 5.24, and taste sense was lowest at 3.69. Therefore, schoolchildren first tend to recognize and deem important the touch and sight of food before its taste.