• Title/Summary/Keyword: Molecular Detection

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Detection of Hydrocarbons to Determine Post-Irradiation of Dry Soup Base Ingredients in Instant Noodle (라면 건조 스프 재료의 방사선 조사 검지를 위한 Hydrocarbon류 검출)

  • Hwang, Keum-Taek;Park, Jun-Young
    • Korean Journal of Food Science and Technology
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    • v.30 no.5
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    • pp.1018-1023
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    • 1998
  • Hydrocarbons in dry soup base and its ingredients in instant noodle were analyzed to determine whether the analysis of hydrocarbons is a potential method to detect post-irradiation of the soup base. Soup base ingredients were irradiated individually, irradiated before mixing, or mixed after irradiation. Lipids were extracted with hexane and hydrocarbons were separated from the lipids through Florisil column. The hydrocarbons were analyzed with GC. Hydrocarbons C17:2, C16:3, C17:1, and C16:2 were detected in palm oil, red pepper powder, and sesame seeds irradiated at 10 kGy, but not in unirradiated ones. C17:2, C16:3, C17:1, and C16:2 were not detected in the soup base mixture of unirradiated ingredients. The four hydrocarbons were detected in the soup base mixture using irradiated palm oil or sesame seeds. In the mixture using irradiated red pepper powder, C17:2 and C16:3 were detected. When the soup mixture was irradiated after mixing unirradiated ingredients, C17:2, C17:1, and C16:2 were detected in the sample irradiated at 1 kGy, and C17:2, C16:3, C17:1, and C16:2 were in large amounts at 5 and 10 kGy.

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One-Stage Polymerase Chain Reaction for the Comprehensive Detection of Type D Retrovirus Provial DNA (Type D Retrovirus 감염의 포괄적 검색을 위한 One-Stage 중합효소 연쇄반응법의 개발)

  • Jeong, Yong-Seok
    • The Journal of Korean Society of Virology
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    • v.27 no.1
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    • pp.19-27
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    • 1997
  • To develop the polymerase chain reaction (PCR) for the detection of type D simian retrovirus (SRV) infection, an oligonucleotide primer pair was designed to hybridize to the sequences within env gene of SRV subtype 1 (SRV-1). The 3' proximal env sequences annealing to the primers had been rather conserved among three different subtypes of SRV, SRV-1, SRV-2, and SRV-3 (Mason-Pfizer Monkey Virus: MPMV). The PCR using the primer pair targeting an env region successfully detected and amplified all three subtypes of SRV with excellent specificity after single round of reaction. The tests with peripheral blood mononuclear cells infected either with simian immunodeficiency virus or simian T-Iymphotropic virus type 1, major immunosuppressive viral agents together with SRV in simian, verified the specificity of the PCR by excluding any cross reactivity. Semiquantitative titration PCR, amplifying serially diluted plasmid DNA of each subtype, was performed to evaluate sensitivity limits of the reaction. Based on molecular weight of each cloned SRV genome, the PCR should be able to detect one SRV-infected cell per more than $5-7{\times}10^4$ uninfected cells after simple ethidium bromide staining of resulting products. The PCR must be very efficient screening system with its quickness, certainty, and sensitivity for SRV-infected animals used in human AIDS research model. Second round amplification of the reaction products from the first PCR, or Southern hybridization by radiolabeled probes shall render to compete its efficacy to ELISA which has been the most sensitive technique to screen SRV infection but with frequent ambiguity problem.

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Department of DNA Chromatographic System for On-Site Detection of Food-Contaminating Bacteria (식중독균 현장탐지를 위한 DNA 크로마토그래피 분석시스템의 개발)

  • 김석하;정우성;백세환
    • KSBB Journal
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    • v.18 no.3
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    • pp.190-196
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    • 2003
  • An analytical system detecting DNA particularly utilizing a concept of membrane strip chromatography initially applied to home-version tests for, such as, pregnancy and ovulation has been developed. We have chosen S. typhimurium as model analyte among food-contaminating microorganisms that occurred in high frequencies, and invA gene, as a detection target, specific to Salmonella species. This gene was able to be amplified by PCR under optimal conditions employing newly designed primers in our laboratory. The PCR product was specifically measured via hybridization between the analyte and a DNA probe, which was a totally different feature from the conventional gel electrophoresis detecting the products based only on the molecular size. It is notable thar the DNA probe sequence was specially designed such that no separation of excess primers present after PCR was required. This was immobilized on a nitrocellulose (NC) membrane via streptavidin-biotin linkage minimizing a steric effect when the hybridization with the amplified DNA took place. The analyrical system detected the microorganism in a concentration of minimum $10^3$ cfu/mL (i.e., 10 cells per system), estimated from the standard curve, 20 to 40 minutes after adding the sample. This sneitivity was approximately 10 times higher than that of gel electrophoresis as an analytical tool conventionally used. Furthermore, the assay was able to be run at room temperature, which would ofter an extra advantage to users.

Purification and Characterization of an Antimicrobial Substance from Bacillus subtilis HH28 Antagonistic to Bacillus cereus (Bacillus cereus를 억제하는 Bacillus subtilis HH28의 항균물질 정제와 특성규명)

  • Cha, Hyun A;Chung, Dawn;Hong, Sung Wook;Chung, Kun Sub
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.393-401
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    • 2014
  • A bacterium producing antimicrobial substance was isolated from cheonggukjang. The bacterium was identified as a strain of Bacillus subtilis by 16S rDNA sequencing and designated as Bacillus subtilis HH28. The antimicrobial substance produced from Bacillus subtilis HH28 was purified by 0-80% ammonium sulfate precipitation, DEAE-sepharose FF column chromatography, and Sephacryl S-200 HR gel chromatography. The molecular weight of the purified antimicrobial substance was estimated to be approximately 3,500 Da using Tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis and direct detection analysis. Antimicrobial substance from B. subtilis HH28 not only inhibited B. cereus, but also Listeria monocytogenes and Vibrio parahaemolyticus. The purified antimicrobial substance was stable at $40-80^{\circ}C$, and between pH 2 and 8. Antimicrobial activity of the purified substance was completely destroyed by treatment of protease, proteinase K, and pronase E, indicating that it is proteinaceous.

Immuno-chromatographic Analysis for HPV-16 and 18 E7 Proteins as a Biomarker of Cervical Cancer Caused by Human Papillomavirus

  • Kim, Joo-Ho;Cho, Il-Hoon;Seo, Sung-Min;Kim, Ji-Sook;Oh, Kyu-Ha;Kang, Heun-Soo;Kim, In-Gyu;Paek, Se-Hwan
    • Bulletin of the Korean Chemical Society
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    • v.30 no.12
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    • pp.2999-3005
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    • 2009
  • Among the more than 120 different types of human papillomavirus (HPV), types 16 and 18 have been known to be high risk agents that cause cervical cancer. We examined, in an immuno-chromatographic analysis, the potential of using the early gene product, E7 protein, as a diagnostic marker of cervical cancer caused by HPV. We developed monoclonal antibodies specific to HPV-16 and 18 E7 proteins that were produced from bacterial cells using gene recombinant technology. For each E7 protein, the optimal antibody pair was selected using the immuno-chromatographic sandwichtype binding system based on the lateral flow through membrane pores. Under these conditions, this rapid testing assay had a detection capability as low as 2 ng/mL of E7 protein. Furthermore, since viral analysis required the host cell to be lysed using chemicals such as detergents, it was possible that the E7 protein was structurally damaged during this process, which would result in a decrease in detection sensitivity. Therefore, we examined the detrimental effects caused by different detergents on the E7 protein using HeLa cells as the host. In these experiments, we found that the damage caused by the detergent, nonylphenylpolyethylene glycol (NP-40), was minimal relative to Triton X-100 commonly used for the cell lysis. Temperature also affected the stability of the E7 protein, and we found that the E7 protein was stabilized at 4$^{\circ}C$ for about 2 h, which was 4 times longer than at room temperature. Finally, a HPV-infected cervical cancer cell line, which was used as a real sample model, was treated using the optimized conditions and the presence of E7 proteins were analyzed by immuno-chromatography. The results of this experiment demonstrated that this rapid test could specifically detect HPV-infected samples.

Study of a Brain Tumor and Blood Vessel Detection System Using Multiple Fluorescence Imaging by a Surgical Microscope (수술현미경에서의 다중형광영상을 이용한 뇌종양과 혈관영상 검출 시스템 연구)

  • Lee, Hyun Min;Kim, Hong Rae;Yoon, Woong Bae;Kim, Young Jae;Kim, Kwang Gi;Kim, Seok Ki;Yoo, Heon;Lee, Seung Hoon;Shin, Min Sun;Kwon, Ki Chul
    • Korean Journal of Optics and Photonics
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    • v.26 no.1
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    • pp.23-29
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    • 2015
  • In this paper, we propose a microscope system for detecting both a tumor and blood vessels in brain tumor surgery as fluorescence images by using multiple light sources and a beam-splitter module. The proposed method displays fluorescent images of the tumor and blood vessels on the same display device and also provides accurate information about them to the operator. To acquire a fluorescence image, we utilized 5-ALA (5-aminolevulinic acid) for the tumor and ICG (Indocyanine green) for blood vessels, and we used a beam-splitter module combined with a microscope for simultaneous detection of both. The beam-splitter module showed the best performance at 600 nm for 5-ALA and above 800 nm for ICG. The beam-splitter is flexible to enable diverse objective setups and designed to mount a filter easily, so beam-splitter and filter can be changed as needed, and other fluorescent dyes besides 5-ALA and ICG are available. The fluorescent images of the tumor and the blood vessels can be displayed on the same monitor through the beam-splitter module with a CCD camera. For ICG, a CCD that can detect the near-infrared region is needed. This system provides the acquired fluorescent image to an operator in real time, matching it to the original image through a similarity transform.

Usefulness of Automated PCR Test for the Detection of Mycobacterium Tuberculosis in Fresh Biopsy Tissues (신선조직 검체에서 결핵균 검출을 위한 자동화 중합효소연쇄반응 검사의 유용성)

  • Choi, Woo Soon;Shin, So Young;Kim, Jong Ok;Kim, Myung Sook;Lee, Hye Kyung
    • Tuberculosis and Respiratory Diseases
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    • v.61 no.1
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    • pp.54-59
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    • 2006
  • Background: Although there have been several studies regarding the clinical value of an automated TB-PCR study using sputum, bronchial washing, and other body fluid samples for the detection of pulmonary tuberculosis, there are only a few reports on the use of fresh tissue samples. Materials and methods: The acid-fast bacilli stain(AFB), tuberculosis culture, automated TB-PCR study, and histopathology examination were performed in 42 fresh tissue samples. Results: Among the 42 cases, 18 cases were diagnosed with tuberculosis based on the clinical findings. Sixteen of the 18 cases were TB-PCR positive and of these 16 cases, only 2 cases were positive in the AFB stain or culture study. However, all 18 cases showed the histopathology findings of chronic granulomatous inflammation that was compatible with tuberculosis. Based on the clinical findings, the sensitivity, specificity, positive predictability, and negative predictability of the automated TB-PCR study were 88.9%, 100%, 100%, and 92.3% respectively. Conclusion: An automated TB-PCR assay is an important diagnostic tool for diagnosing tuberculosis in fresh tissue samples.

Optimum Extraction Methods of Volatile Compounds in Beef Extract Powder (쇠고기 엑기스 분말 휘발성 성분의 최적 추출방법에 관한 연구)

  • Kim Hun;Cho Woo-Jin;Jeong Eun-Jeong;Ahn Jun-Suck;Lim Chi-Won;Yoo Young-Jae;Kim Kwang-Ho;Cha Yong-Jun
    • The Korean Journal of Food And Nutrition
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    • v.17 no.4
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    • pp.412-419
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    • 2004
  • In odor to select optimum extraction methods of volatile compounds in beef extract powder(BEP) as basic data for the development of a new detection method of irradiated BEP, four extraction methods, such as solid phase microextraction with polar fiber(S-PD) and non-polar fiber(S-CD), purge and trap(P&T) and liquid liquid continuous extraction(LLCE) methods, were tested with gas chromatography/mass spectrometry method. A total of 106 volatile compounds including 22 hydrocarbons, 7 aldehydes, 6 ketones, 13 alcohols, 6 sulfur-containing compounds, 19 nitrogen-containing compounds, 6 aromatic compounds, 17 terpenes, 8 furans and 2 miscellaneous compounds were detected in BEP by four detection methods. The most compounds(62 compounds) were detected by S-PD method, followed by P&T(43), LLCE(38) and S-CD method(30). Among these methods, S-PD and P&T methods showed a complementary interrelationship to detect volatile compounds as S-PD method showed high detectabiltiy to all compound groups except hydrocarbons and ketones, which had high volatility and low molecular weight(less than RI 1200), but P&T method showed the contrary pattern to that of S-PD method. Moreover, the most of volatile compounds detected by S-CD and LLCE methods were also detectable by S-PD or/and P&T methods. Therefore, the simultaneous application of S-PD and P&T methods were selected as the optimum volatile extraction methods of BEP.

Detection of Post-Irradiation of Dry Soup Base Ingredients in Instant Noodle by Thermoluminescence Technique (라면 건조 수프 재료의 방사선 조사 검지를 위한 Thermoluminescence 활용)

  • Hwang, Keum-Taek;Byun, Myung-Woo;Wagner, Ute;Dehne, Lutz I.
    • Korean Journal of Food Science and Technology
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    • v.30 no.4
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    • pp.759-766
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    • 1998
  • It was determined whether thermoluminescence (TL) technique is a potential method to detect post-irradiation of dry soup base mix for instant noodle and its ingredients. The ingredients or the mixtures were irradiated, from which minerals were isolated using sodium polytungstate solution. Their TL signals (1st glow TL intensity/2nd glow TL intensity reirradiated at 48.6 Gy) were measured. The TL signals in temperature ranges of $229{\sim}295^{\circ}C$ and $229{\sim}361^{\circ}C$ showed larger differences between unirradiated and irradiated samples compared to other ranges. The average TL signals for unirradiated garlic powder, ginger powder, black pepper powder, onion powder, red pepper powder, and sesame seeds were below 0.2, while those for onion powder, red pepper powder, and sesame seeds irradiated at 10 kGy were over 20 in the two temperature ranges. The average TL signals for unirradiated soup mixture were 0.08 and 0.1, respectively, in the two temperature ranges, while those for the mixtures containing 10 kGy-irradiated onion powder, red pepper powder, and sesame seeds individually or in combination were over 7. The average TL signals for the mixtures irradiated 1, 5, and 10 kGy were over 10.

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Molecular epidemiologic trends of norovirus and rotavirus infection and relation with climate factors: Cheonan, Korea, 2010-2019 (노로바이러스 및 로타바이러스 감염의 역학 및 기후요인과의 관계: 천안시, 2010-2019)

  • Oh, Eun Ju;Kim, Jang Mook;Kim, Jae Kyung
    • Journal of Digital Convergence
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    • v.18 no.12
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    • pp.425-434
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    • 2020
  • Background: Viral infection outbreaks are emerging public health concerns. They often exhibit seasonal patterns that could be predicted by the application of big data and bioinformatic analyses. Purpose: The purpose of this study was to identify trends in diarrhea-causing viruses such as rotavirus (Gr.A), norovirus G-I, and norovirus G-II in Cheonan, Korea. The identified related factors of diarrhea-causing viruses may be used to predict their trend and prevent their infections. Method: A retrospective analysis of 4,009 fecal samples from June 2010 to December 2019 was carried out at Dankook University Hospital in Cheonan. Reverse transcription-PCR (RT-PCR) was employed to identify virus strains. Information about seasonal patterns of infection was extracted and compared with local weather data. Results: Out of the 4,009 fecal samples tested using multiplex RT-PCR (mRT-PCR), 985 were positive for infection with Gr.A, G-I, and G-II. Out of these 985 cases, 95.3% (n = 939) were under 10 years of age. Gr.A, G-I, and G-II showed high infection rates in patients under 10 years of age. Student's t-test showed a significant correlation between the detection rate of Gr.A and the relative humidity. The detection rate of G-II significantly correlated with wind-chill temperature. Conclusion: Climate factors differentially modulate rotavirus and norovirus infection patterns. These observations provide novel insights into the seasonal impact on the pathogenesis of Gr.A, G-I, and G-II.