• Title/Summary/Keyword: Reaction monitoring

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Rapid Identification of Lactobacillus plantarium in Kimchi Using Polymerase Chain Reaction

  • Kim, Tae-Woon;Min, Sung-Gi;Choi, Dong-Hun;Jo, Jae-Sun;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.881-884
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    • 2000
  • A polymerase chain reaction (PCR) was performed to rapidly identify Lactobacillus plantarum from type strains and kimchi samples. The PCR experiments were carried out using specific oligonucleotide primer sets based on the 16S rRNA gene sequences of L. plantarum. The expected DNA amplificate of 419 bp was obtained when either purified DNA or whole cells of L. plantarum strains reacted with LP primers, yet not with any of the other strains. The PCR product was confirmed by DNA sequencing. Accordingly, since the PCR method used is simple, specific, and rapid, it will be useful for monitoring and evaluation L. plantarum in the mixed microbial population found in kimchi.

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Imaging the Enzymatic Reaction of Urease Using Liquid Crystal-Based pH Sensor

  • Hu, Qiong-Zheng;Jang, Chang-Hyun
    • Bulletin of the Korean Chemical Society
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    • v.32 no.12
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    • pp.4377-4381
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    • 2011
  • In this study, real-time and label-free methods for monitoring the enzymatic reaction of urease, which releases ammonia through the hydrolysis of urea in an aqueous solution, were developed using a liquid crystal (LC)-based pH sensor. Nematic liquid crystal 4-cyano-4'-pentylbiphenyl (5CB), doped with 4'-pentyl-biphenyl-4-carboxylic acid (PBA), exhibited a shift in optical appearance from bright to dark when it was in contact with ammonia generated from the enzymatic reaction between urease and urea. This optical change was attributed to the anchoring transitions of LCs caused by hydrophobic interactions between the tails of deprotonted PBA ($PBA^-$) molecules and the LCs at the aqueous/LC interface. This novel technique holds great promise for the sensitive detection of urease along with its substrates and inhibitors.

Detection of Coliform and Escherichia coli in Spring Water by Polymerase Chain Reaction (PCR법을 이용한 옹달샘물의 대장균군 및 대장균 검출)

  • 류승희;박석기
    • Journal of Environmental Health Sciences
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    • v.28 no.2
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    • pp.193-202
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    • 2002
  • The polymerase chain reaction(PCR) of target lacZ and uidA genes were used to detect total coliform and Escherichia coli for determining water quality, respectively. Of 109 spring waters, coliform were detected from 38 spring waters by lacZ PCR method but 21 spring waters by culture method accepted by the Ministry of Environment for water quality monitoring. The lacz PCR method gave the results statistically equivalent to those of the culture method(kappa=0.62, McNemar=17.00). The uidA PCR method gave the same results to those of the culture method. The sensitivity and specificity of coliform and E. coli by PCR method were 100% and 80.7%, respectively. Therefore, PCR can be used for the rapid identification of Escherichia coli and coliform in potable water using uidA and lacZ.

CRDS Study of Tropospheric Ozone Production Kinetics : Isoprene Oxidation by Hydroxyl Radical

  • Park, Ji-Ho
    • Journal of Environmental Health Sciences
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    • v.35 no.6
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    • pp.532-537
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    • 2009
  • The tropospheric ozone production mechanism for the gas phase additive oxidation reaction of hydroxyl radical (OH) with isoprene (2-methyl-1,3-butadiene) has been studied using cavity ring-down spectroscopy (CRDS) at total pressure of 50 Torr and 298 K. The applicability of CRDS was confirmed by monitoring the shorter (~4%) ringdown time in the presence of hydroxyl radical than the ring-down time without the photolysis of hydrogen peroxide. The reaction rate constant, $(9.8{\pm}0.1){\times}10^{-11}molecule^{-1}cm^3s^{-1}$, for the addition of OH to isoprene is in good agreement with previous studies. In the presence of $O_2$ and NO, hydroxyl radical cycling has been monitored and the simulation using the recommended elementary reaction rate constants as the basis to OH cycling curve gives reasonable fit to the data.

Analysis of Vitellogenin Gene Expression in Synechogobius hastus (Gobiidae) (풀망둑 난황전구단백질 유전자발현 추적기법)

  • 계명찬
    • Korean Journal of Environmental Biology
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    • v.22 no.1
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    • pp.206-212
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    • 2004
  • In an effort to develop the tools for monitoring the contamination of xenoestrogen in the aquatic environment of Korea, reverse transcription-polymerase chain reaction (RT-PCR) analysis of vitellogenin (VTG) mRNA expression were optimized in Synechogobius hastus. Based on the partial VTG cDNA sequence VTG mRNA level in livers from male fishes was analyzed by RT-PCR. As an internal control beta actin mRNA was amplified. 3 ${\mu}g$ of total RNA was reverse transcribed in 20 $\mu$l reaction using murine leukemia virus 〔MuLV〕 reverse transcriptase. Subsequent PCR using the 1 ${\mu}g$ of cDNA resulted in linear increase in PCR product of VTG in female liver cDNA from 10 to 30 cycles of amplification. On the contrary, in male, PCR product first detected at 28 cycles of amplification and linearly increased during 38 cycles of amplification, suggesting that male S. hastus expresses minute amount of VTG mRNA which is $2^{-18}$ equivalent of female. In conclusion, the optimized protocol of VTG mRNA expression in the liver of male S. hastus will be promising the environmental monitoring the xenoestrogen contamination in the western coast and estuaries in Korea.

Rapid Screening of Phospholipid Biomarker Candidates from Prostate Cancer Urine Samples by Multiple Reaction Monitoring of UPLC-ESI-MS/MS and Statistical Approaches

  • Lim, Sangsoo;Bang, Dae Young;Rha, Koon Ho;Moon, Myeong Hee
    • Bulletin of the Korean Chemical Society
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    • v.35 no.4
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    • pp.1133-1138
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    • 2014
  • Ultrahigh performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI- MS/MS) provides a high-speed method to screen a large number of samples for small molecules with specific properties. In this study, UPLC-ESI-MS/MS with multiple reaction monitoring (MRM) was employed to screen urinary phospholipid (PL) content for biomarkers of prostate cancer. From lists of urinary PLs structurally identified using nanoflow LC-ESI-MS/MS, 52 PL species were selected for quantitative analysis in urine samples between 22 cancer-free urologic patients as controls and 45 prostate cancer patients. Statistical treatment of data by receiver operating characteristic (ROC) analysis yielded 14 PL species that differed significantly in relative concentrations (area under curve (AUC) > 0.8) between the two groups. Among PLs present at higher levels in prostate cancer urine, phosphatidylcholines (PCs) and phosphatidylinositols (PIs) constituted the major head group PLs (3 PCs and 7 PIs). For technical reasons, PL species of low abundance may be underrepresented in data from UPLC-ESI-MS/MS performed in MRM mode. However, the proposed method enables the rapid screening of large numbers of plasma or urine samples in the search for biomarkers of human disease.

Adverse Drug Reaction Surveillance System in Korea (우리나라 약물유해반응 감시체계)

  • Choi, Nam-Kyong;Park, Byung-Joo
    • Journal of Preventive Medicine and Public Health
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    • v.40 no.4
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    • pp.278-284
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    • 2007
  • Despite extensive researches and pre-market clinical trials, only limited information on the adverse drug reactions (ADRs) of a drug can be collected at the time of market approval from regulatory agency. ADRs constitute a major public health problem. Post-marketing surveillance of drugs is important to detect signals for ADR. In Korea, one of the main methods for monitoring the safety of marketed drugs is spontaneous reporting system of suspected ADRs. Re-examination and re-evaluation system are in force for monitoring safety of new market approval drugs and currently under marketing drugs, respectively. Recently, regional pharmacovigilance centers were designated from Korean Food and Drug Administration for facilitating ADR surveillance. Over recent years, with the development of information technology, there has been an increased interest in establishing data mining system for detecting signals from Health Insurance Review Agency database. The purpose of this paper is to review the current status of Korean ADR surveillance system and suggest the possible solutions for developing active pharmacovigilance system in Korea.

Development and Evaluation of a SYBR Green-Based, Real-time Polymerase Chain Reaction for Rapid and Specific Detection of Human Coxsackievirus B5

  • Cho, Kyu Bong
    • Biomedical Science Letters
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    • v.26 no.4
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    • pp.302-309
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    • 2020
  • Human Coxsackievirus B5 (HuCoxV-B5) infection has been associated with various diseases such as myocarditis, aseptic meningitis, hand-foot-and mouth-disease, and insulin-dependent diabetes. HuCoxV-B5 is a virus transmitted through the fecal-oral route and is detected in clinics, aquatic environments, food, shellfish, etc. and is one of the more important viruses in public health because of its incidence rate reported worldwide. In this study, a combination of SYBR Green-based real-time PCR primers for molecular diagnosis including monitoring of HuCoxV-B5 was selected and the optimal reaction conditions were established. Compared with the previously reported TaqMan probe-based real-time PCR method, assessments including a sample applicability test were performed. Results showed that the real-time PCR method developed in this study was suitable for a molecular diagnostic technique for detecting HuCoxV-B5. This study is expected to contribute to efforts in responding to safety accidents in public health because the proposed method facilitates rapid diagnosis of clinical patients. It can also be used as a specific monitoring tool of HuCoxV-B5 in non-clinical areas such as aquatic environments among others.

Genetic information analysis for the development of an event-specific PCR marker for herbicide tolerance LM crops

  • Do Yu, Kang;Myung Ho, Lim;Soo In, Sohn;Hyun Jung, Kang;Tae Sung, Park
    • Korean Journal of Agricultural Science
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    • v.48 no.4
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    • pp.1051-1065
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    • 2021
  • Recent times have seen sustained increases in genetically modified (GM) crops not only for cultivation but also for the utility of food and feed worldwide. Domestically, commercial planting and the accidental or unintentional release of living modified (LM) crops into the environment are not approved. Many detection methods had been devised in an effort to realize effective management of the safety of agricultural genetic resources. In order to develop event-specific polymerase chain reaction (PCR) markers for LM crops, we analyzed the genetic information of LM crops. Genetic components introduced into crops are of key importance to provide a basis for the development of detection methods for LM crops. To this end, a total of 18 varieties from four major LM crop species (maize, canola, cotton, and soybeans) were subjected to an analysis. The genetic components included introduced genes, promoters, terminators and selection markers. Thus, if proper monitoring techniques and single or multiplex PCR strategies that rely on selection markers can be established, such an accomplishment can be regarded as a feasible solution for the safe management of staple crop resources.