• 제목/요약/키워드: KIT-6

검색결과 804건 처리시간 0.024초

인삼포장에서 뿌리섞음병원균의 진단을 위한 RT-PCR KIT의 개발 (Development of RT-PCR Kit for Diagnosis of Pathogenic Agent of Ginseng Root Rot in the Ginseng Field)

  • 도은수
    • 한국자원식물학회지
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    • 제16권1호
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    • pp.40-48
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    • 2003
  • C. destructans는 인삼에서 가장 문제가 되고 있는 뿌리섞음병을 유발하는 매우 중요한 미생물이다. 현재까지 정상적인 인삼포장이나 폐포지에서도 이 병원균의 농도를 조사할 만한 방법이 없어 이를 쉽게 조사함으로서 인삼 예정지 관린시 도움을 줄 수 있는 새로운 방법이 절실이 요구되고 있다. 본 연구에서는 nested PCR이란 분자생물학적 방법을 이용하여 효과적으로 매우 낮은 농도의 C. destructans을 검출할 수 있는 방법을 개발하였다. 2개의 universal ITS primers(ITS5F와 ITS4R)을 사 용 하 여 Cylindrocarpon spp.의 rDNA로부터 ITS영역을 증폭하였다. 이어 C. destructans의 specific primer(Nest 1 과 Nest 2)을 사용하여 최적의 PCR조건으로 재증폭시켜 밴드를 확인하였다. 또한 이런 2번의 과정을 4개의 primer를 동시에 사용함으로서 한번에 확인할 수 있는 방법을 개발하였으며 이에 따른 PCR조건도 확립하였다. 따라서 본 방법에 의해서 인삼포장의 토양에서 채취된 매우 낮은 농도의 wild type C. destructans spore로부터 성공적으로 positive band을 확인함으로써 추후 인삼포장의 선정 및 4년생에서 6년까지(홍삼포) 재배기간등의 예측에 활용 될 것으로 생각된다.

Magnolol과 Honokiol이 항균, 교원질 분해효소, 세포독성 및 Cytokine생산에 미치는 영향 (THE EFFECTS OF HONOKIOL AND MAGNOLOL ON THE ANTIMICROBIAL, BACTERIAL COLLAGENASE ACTIVITY, CYTOTOXICITY AND CYTOKINE PRODUCTION)

  • 장범석;손성희;정종평;배기환
    • Journal of Periodontal and Implant Science
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    • 제23권1호
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    • pp.145-158
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    • 1993
  • The oral microbiota such as P. gingivalis, P. intermedia and A. actinomycetemcomitans play a primary role in the initiation and progression of the periodontal disease. The purpose of this study was to evaluate the antimicrobial effects and inhibitory effects of honokiol and magnolol on the bacterial collagenase activity, cytotoxicity and cytokine production of periodontopathic microorganisms. The antimicrobial activities of honokiol and magnolol was evaluted with minimum inhibition concentration. Honokiol was more active than magnolol, but less than chlorhexidine on antimicrobial activity. The inhibitory effects of magnolol and honokiol on the collagenolytic activity and cytotoxicity were evaluated using a Collagenokit CLN-100 and rapid colorimetric assay (MTT method) for cellular growth and survival of gingival fibroblast and periodontalligament cell and $[^3H]-thymidine$ incorporation for the gingival epithelial cell. The inhibitory effects on the collagenolytic activity was the highest in chlorhexidine, and the lowest in magnolol. Magnolol had the lowest cytotoxic effect and chlorhexidine had the highest. The inhibitory effects on cytokine production was evaluated using $interleukin-1{\beta}$ ELISA kit (Cistron Biotech.), IL-6, $TNF-{\alpha}$ ELISA kit (Genzyme) and inhibitory effects were higher than bacterial LPS and there is no difference among the honokiol, magnolol and chlorhexidine. From these results, the antimicrobial and antienzymatic activities of honokiol and magnolol were seemed to inhibit bacterial growth and enzyme activities with lesser cytotoxic activities. Therefore, it was suggested that honokiol and magnolol are very effective antimicrobial agents on periodontal pathogens.

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마 에탄올추출물의 피부 collagen 합성 촉진 및 MMPs 활성 억제효과 (Ethanol Extract of Dioscorea batatas Stimulates Procollagen Production and Reduces UVB-induced MMPs Activity in Skin)

  • 김대성;전병국;임난영;문연자;이영은;우원홍
    • 동의생리병리학회지
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    • 제27권2호
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    • pp.183-188
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    • 2013
  • Ultraviolet (UV) B irradiation induces the production of matrix metalloproteinases (MMPs), which are responsible for the degradation or synthesis inhibition of collagenous extracellular matrix in connective tissues, causing skin photoaging. In this study, we examined the inhibitory effect of MMP-1 expression of yam extract in tumor necrosis factor-alpha (TNF-${\alpha}$)-stimulated human dermal fibroblast neonatal (HDFn) cell and preventive effect of UVB-induced damage in hairless mice skin. The synthesis of procollagen and the release of MMP-1 in HDFn cells were measured by EIA kit and MMP-1 assay kit, respectively. UVB radiation was applied to the backs of the mice three times a week for 8 weeks. Mice were randomly divided into three groups, and were topical application with the Dioscorea batatas (DB, 6%) or vehicle. Reduction of TNF-${\alpha}$-induced procollagen synthesis was increased by DB (50 ug/ml), which was higher than positive control group (TGF-${\beta}$). Also, pre-treatment of HDFn cells with DB inhibited TNF-${\alpha}$-induced release of MMP-1. In vivo study, we found that preventive effect of DB against UV-induced epidermal thickness. DB suppressed the expression of MMP-3 and MMP-13 induced by UVB irradiation. Our results show that DB have preventive effect of UV-induced skin damage in hairless mice.

가축 소화기 병원성 세균을 저해하는 유산균의 분리 및 동정 (Isolation and Identification of Lactic Acid Bacteria Inhibiting Gastro-intestinal Pathogenic Bacteria of Domestic Animal.)

  • 이재연;황교열;김현수;김근;성수일
    • 한국미생물·생명공학회지
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    • 제30권2호
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    • pp.129-134
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    • 2002
  • 토종닭 소장으로부터 가축 소화기 병원성 유해세균의 분리를 위하여 무작위 선별법과 중층-도말법을 사용하여 총130주의 유산균을 1차 분리하였다. 1차 분리한 130주를 paper disc법을 사용하여 최종적으로 살모넬라 균, 포도상구균, 대장균에 대하여 저해능이 우수한 7주를 분리하였다. 분리 균주의 동정을 위하여 1차로 API CHL kit를 사용하였고, 동정의 정확성을 높이기 위하여 2차로 16S rRNA sequencing 방법을 사용하였다. 그 결과 BD14 균주는 Pediococcus pentosaceus로 동정되었으며 나머지 6주는 Lactobacillus sp.로 동정되어 모두 안전성 있는 유산균임을 확인하였다. 여러 병원균에 가장 항균력이 큰 유산균은 L. pentosus K34이었으며, 이 유산균은 염산과 담즙산에 대한 내성에서도 가장 우수한 균주로 나타났다. 10종의 항생제에 대한 감수성을 조사한 결과 ciprofloxacin에 대하여 모든 균주가 내성을 보였고, BL 균주를 제외하고 모든 균주가 colistin, streptomycin에 대하여 내성을 보였다. 또한 BD14, BD16, K34 균주가 gentamicin에 대한 내성을 보였다.

Screening of Lactobacilli Derived from Chicken Feces and Partial Characterization of Lactobacillus acidophilus A12 as Animal Probiotics

  • Lee, Na-Kyoung;Yun, Cheol-Won;Kim, Seung-Wook;Chang, Hyo-Ihl;Kang, Chang-Won;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • 제18권2호
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    • pp.338-342
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    • 2008
  • This study was performed to screen and select Lactobacillus strains from chicken feces for probiotic use in animals. Of these strains, strain AU had the highest immunostimulatory effect. Therefore, strain A12 was characterized as a potential probiotic. Strain A12 was tentatively identified as Lactobacillus acidophilus A12, using the API 50 CHL kit based on a 99.9% homology. L. acidophilus A12 was highly resistant to artificial gastric juice (pH 2.5) and bile acid (oxgall). Based on results from the API ZYM kit, leucine arylamidase, crystine arylamidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase, ${\alpha}$-galactosidase, ${\beta}$-galactosidase, ${\alpha}$-glucosidase, ${\beta}$-glucosidase, and N-acetyl-${\beta}$-glucosamidase were produced by strain A12. L. acidophilus A12 showed resistance to several antibiotics (nisin, gentamicin, and erythromycin). The amount of interleukin $(IL)-1{\alpha}$ in $20{\times}$ concentrated supernatant from L. acidophilus A12 was approximately 156pg/ml. With regard to antioxidant activity, L. acidophilus A12 supernatant showed 60.6% DPPH radical scavenging activity. These results demonstrate the potential use of L. acidophilus A12 as health-promoting probiotics.

Comparison of Ogawa Media, BACTEC MGIT 960 System and TB/NTM Real-Time PCR for Detecting Mycobacterium Species

  • Bang, Hae-In;Choi, Tae-Youn;Shin, Jeong-Won
    • Tuberculosis and Respiratory Diseases
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    • 제71권4호
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    • pp.249-253
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    • 2011
  • Background: Mycobacterial infection is a problem throughout the world along with the increase of immunocompromised patients. For this reason, there have been many methods for faster and more accurate diagnosis. In this study, we evaluated several laboratory methods for mycobacterial infection. Methods: From January to December 2009, 635 specimens were cultured with mycobacteria growth indicator tube (MGIT) and Ogawa media. Polymerase chain reaction (PCR) was performed with the AdvanSure tuberculosis (TB)/non-tuberculosis mycobacterium (NTM) real-time PCR Kit (LG Life Sciences, Seoul, Korea). The 69 samples showing positive culture results were identified with the AdvanSure Mycobacteria Genotyping Chip Kit (LG Life Science, Seoul, Korea). Results: Sixty-nine (10.9%) out of 635 samples showed positive results for mycobacterial culture. Among the 635 samples, 64 were positive in MGIT, but only 42 were positive in Ogawa media. Of the 635 samples, 607 (95.6%) showed the same results between MGIT and Ogawa and the results of 579 (95.4%) were also consistent with the TB/NTM real-time PCR results. However, in the case of NTM, only one (1/24, 4.2%) was positive in PCR. In the Mycobacteria genotyping chip analysis, the most frequently identified NTM species in descending order were M. avium, M. intracellulare, M. chelonae and M. abscessus. Conclusion: Culturing with a combination of MGIT and Ogawa is recommended to increase the recovery rate of mycobacteria. Although PCR missed a reasonable number of NTM, it is faster and usually gives results that concur with those from the culture. The appropriate combination of diagnostic methods with clinical correlation are necessary.

Quality Evaluation of Five Commercial Enzyme Linked Immunosorbent Assay Kits for Detecting Aflatoxin B1 in Feedstuffs

  • Sun, Dan-Dan;Gu, Xu;Li, Jun-Guo;Yao, Ting;Dong, Ying-Chao
    • Asian-Australasian Journal of Animal Sciences
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    • 제28권5호
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    • pp.691-696
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    • 2015
  • The objective of this study was to evaluate the quality of five commercial enzyme linked immunosorbent assay (ELISA) kits (A, B, C, D, and E) from different suppliers for detecting aflatoxin $B_1$ ($AFB_1$). $AFB_1$-free corn samples supplemented with different levels of $AFB_1$ (5, 10, and $20{\mu}g/kg$) were used as positive controls and 6 replicates of each control sample were tested to evaluate the accuracy and precision of these kits. In addition, we also evaluated the performance of these ELISA kits for $AFB_1$ in 30 feed samples, including corn, distillers dried grains with soluble, wheat samples, soybean meal, and poultry feed, which were verified by high performance liquid chromatography. Results showed that the coefficients of variation ranged from 1.18% to 16.22% in intra-plate and 2.85% to 18.04% in inter-plate for the determination of $AFB_1$. The half maximal inhibitory concentration for five kits ranged from 3.72 to $7.22{\mu}g/kg$. The quantitation limits of $AFB_1$ were all under the legal limit in China but somewhat inconsistent with kit instructions. Although the recovery rate of four of the five kits were either less than 90% or more than 110%, all these values were acceptable in practice. Two kits had high false positive rates (C and E). In conclusion, our results revealed that the qualities of five tested ELISA kits were significantly different.

Pioneering Annual Colorectal Cancer Screening and Treatment Targeting Low Income Communities in Malaysia (2010-2015)

  • Tze, Christina Ng Van;Fitzgerald, Henry;Qureshi, Akhtar;Tan, Huck Joo;Low, May Lee
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권7호
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    • pp.3179-3183
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    • 2016
  • The aim of this study was to assess the rate of uptake of a customised annual Colorectal Cancer Awareness, Screening and Treatment Project (CCASTP) using faecal immunohistochemical test (FIT) kits in low income communities in Malaysia. The immediate objectives were (1) to evaluate the level of adherence of CRC screening among low-income groups, (2) to assess the knowledge and awareness of the screened population and (3) to assess the accuracy of FIT kits. A total of 1,581 FIT kits were distributed between years 2010 to 2015 to healthy asymptomatic participants of the annual CCASTP organized by Empowered - the Cancer Advocacy Society of Malaysia. Data for socio-demographic characteristics, critical health and lifestyle information of the registered subjects were collected. Findings for use of the FIT kits were collected when they were returned for stool analyses. Those testingd positive were invited to undergo a colonoscopy examination. A total of 1,436 (90.8%) of the subjects retuned the FIT-kits, showing high compliance. Among the 129 subjects with positive FIT results, 92 (71.3%) underwent colonoscopy. Six cases (6.5%) of CRC were found. Based on the data collected, the level of awareness of stool examination and knowledge about CRC was poor amongst the participants. Gender, age group, ethnicity and risk factors (i.e. smoking, lack of exercise and low consumption of fresh fruits) were associated with positive FIT-kit results. In conclusion, CRC screening can be performed in the community with a single FIT-kit. Although CRC knowledge and awareness is poor in low-income communities, the average return rate of the FIT kits and rate of colonoscopy examination were 91.2% and 70.3%, respectively.

Prevelance of Common YMDD Motif Mutations in Long Term Treated Chronic HBV Infections in a Turkish Population

  • Alagozlu, Hakan;Ozdemir, Ozturk;Koksal, Binnur;Yilmaz, Abdulkerim;Coskun, Mahmut
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권9호
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    • pp.5489-5494
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    • 2013
  • In the current study we aimed to show the common YMDD motif mutations in viral polymerase gene in chronic hepatitis B patients during lamivudine and adefovir therapy. Forty-one serum samples obtained from chronic hepatitis B patients (24 male, 17 female; age range: 34-68 years) were included in the study. HBV-DNA was extracted from the peripheral blood of the patients using an extraction kit (Invisorb, Instant Spin DNA/RNA Virus Mini Kit, Germany). A line probe assay and direct sequencing analyses (INNO-LIPA HBV DR v2; INNOGENETICS N.V, Ghent, Belgium) were applied to determine target mutations of the viral polymerase gene in positive HBV-DNA samples. A total of 41 mutations located in 21 different codons were detected in the current results. In 17 (41.5%) patients various point mutations were detected leading to lamivudin, adefovir and/or combined drug resistance. Wild polymerase gene profiles were detected in 24 (58.5%) HBV positive patients of the current cohort. Eight of the 17 samples (19.5%) having rtM204V/I/A missense transition and/or transversion point mutations and resistance to lamivudin. Six of the the mutated samples (14.6%) having rtL180M missense transversion mutation and resistance to combined adefovir and lamivudin. Three of the mutated samples (7.5%) having rtG215H by the double base substituation and resistance to adefovir. Three of the mutated samples (7.5%) having codon rtL181W due to the missense transversion point mutations and showed resistance to combined adefovir and lamivudin. Unreported novel point mutations were detected in the different codons of polymerase gene region in the current HBV positive cohort fromTurkish population. The current results provide evidence that rtL180M and rtM204V/I/A mutations of HBV-DNA may be associated with a poor antiviral response and HBV chronicity during conventional therapy in Turkish patients.

치아인접면 삭제 및 연마에 따른 법랑질 표면의 변화에 관한 주사전자현미경적 연구 (A SCANNING ELECTRON MICROSCOPIC STUDY ON CHANGES OF ENAMEL SURFACE BY INTERPROXIMAL STRIPPING AND POLISHING)

  • 김영순;김광원
    • 대한치과교정학회지
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    • 제22권1호
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    • pp.123-131
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    • 1992
  • This study was done to evaluate the changes of enamel surface by interproximal stripping and recovery of it by polishing. The number of 34 1st premolars which had extracted for orthodontic treatment were selected as samples. Interproximal stripping was performed by hand with metal strip and strip placer (Dentaurum Co., Germany) and low speed handpiece with diamond disk (Superdiaflex, Germany). Polishing was performed by hand with plastic strip (3M Col) and low speed handpiece with whip-mix, DCPA (Dicalcium Phosphate, Anhydrous, $CaHPO_4$) powder and Sof-lex (3M Co. U.S.A.) polishing kit. Each groups were examined under the scanning electron microscope (JEOL Co., JSM-840A, Japan) and the following results were obtained: 1. The stripped group performed by metal strip and diamond disk altogether showed deep furrow on the enamel surface as wide as about $10{{\mu}m}$. 2. There could be seen more irregular scratched line in the group stripped by metal strip than that by diamond disk. 3. The polished group performed by plastic strip and DCPA powder showed slight smoothening of the edge of stripped furrow on the enamel surface without relation to the stripping method. 4. The polished group performed by Sof-lex progressive polishing kit could not avoid the formation of the furrows on the enamel surface according to the particle size without relation to the stripping method. 5. The polished group performed by the superfine polishing wheel, the final stage of Sof-lex polishing method showed shallow scratched line as wide as within about $2{{\mu}m}$ on the enamel surface without relation to the stripping method. 6. The interproximal stripped enamel surface could not recover its original surface texture by any kind of polishing methods.

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