• Title/Summary/Keyword: rapid detection method

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Detection of Pyrazinamide Resistance in Mycobacterium Tuberculosis by Sequencing of pncA Gene (pncA 유전자의 염기 서열 결정에 의한 결핵균의 Pyrazinamide 내성 진단)

  • Hwang, Jee-Yoon;Kwak, Kyung-Rok;Park, Hye-Kyung;Lee, Ji-Seok;Park, Sam-Seok;Kim, Yun-Seong;Lee, Jung-Yoo;Chang, Chul-Hun;Lee, Min-Ki;Park, Soon-Kew
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.1
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    • pp.94-105
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    • 2001
  • Background : Examining the biological susceptibility of Mycobacterium tuberculosis to pyrazinamide (PZA) in vitro is very difficult as PZA is inactive under normal culture conditions. The biological susceptibility test, an enzyme assay for Pzase activity, and a genetic test for pncA gene mutations, were performed in order to predict PZA resistance. Methods : 28 cultured clinical isolates of Mycobacterium tuberculosis were tested. The biological susceptibility was performed by the absolute concentration method using Lowenstein-Jensen media. The PZase activity was tested by means of Wayne's method. A 710-bp region includes the entire open reading frame of pncA was amplified and sequenced. Results : All six strains with positive PZase activity exhibited no pncA mutations with one strain showing a false resistance in the biological susceptibility test. Among the 22 strains with no PZase activity, 21 exhibited showed pncA mutations. In the biological susceptibility test, 20 strains were resistant, and one was susceptible, and the other flied to test. The mutation types varied with ten missense, one silent and one nonsense mutation 1 slipped-strand mispairing, and 6 frameshift mutations. Three strains had an adenine to guanine mutation at position -11 upstream of the start codon. Conclusion : The mutation at the pncA promotor region is frequent at -11 upstream position. Automatic sequencing of pncA is a useful tool for rapid and accurate detection of PZA resistant M. tuberculosis, and for demonstrating the epidemiological relatedness of the PZA resistant M. tuberculosis strains.

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Establishment of a Method for the Analysis of Diarrhetic Shellfish Poisoning by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS를 이용한 설사성패류독소의 분석조건 확립)

  • Lee, Ka-Jeong;Suzuki, Toshiyuki;Kim, Poong-Ho;Oh, Eun-Gyoung;Song, Ki-Cheol;Kim, Ji-Hoe
    • Korean Journal of Food Science and Technology
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    • v.41 no.4
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    • pp.458-463
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    • 2009
  • To establish and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the rapid and accurate quantitation of diarrhetic shellfish poisoning (DSP) toxins, we compared the results from different mobile phases and columns used for their analysis and collision energies for MS/MS experiments. Clear peaks of okadaic acid (OA) and dinophysistoxin-1 (DTX1) were obtained by using a mobile phase comprising aqueous acetonitrile containing 2 mM ammonium formate and 50 mM formic acid. The collision energies were optimized to facilitate the most sensitive detection for each toxin, namely, OA, DTX1, pectenotoxin-2 (PTX2), or yessotoxin (YTX). Further, the maximum ion response was obtained at a collision energy of 48 V for OA and DTX1. We compared the analytical performance of $C_8$ and $C_{18}$ columns. A wide range of toxins namely, OA, DTX1, PTX2, and YTX, except DTX3, were detected by both the columns. Although DTX3 was only detected by the $C_8$ column, we found that the $C_{18}$ column was also suitable for the quantitation of OA and DTX1 the toxins responsible for inducing diarrhea. The limit of detection of OA and DTX1 by the established LC-MS/MS conditions was 1 ng/g, and the limit of quantitation of the toxins under the same conditions was 3 ng/g. The process efficiencies were 91-118% for oysters (Crassostrea gigas) and 96-117% for mussels (Mytilus galloprovincialis) further, we observed no significant effect of matrix during the ionization process in LC-MS/MS. The comparison between mouse bioassay (MBA) and LC-MS/MS yielded varying results because low concentrations of OA and DTX1 were detected by LC-MS/MS in some shellfish samples, which provided positive results on MBA for DSP. The analysis time required by MBA for DSP analysis can be reduced by LC-MS/MS.

Determination of Total Aflatoxins in Foods by Parallelism of ELISA and LC/MS/MS (ELISA-LC/MS/MS 병행에 의한 식품 중 aflatoxins 분석)

  • Kim, Kyeong-Yeol;Nam, Min-Ji;Nam, Bo-Ram;Ryu, Hee-Jung;Song, Jeong-Eon;Shim, Won-Bo;Lee, Soo-Hyung;Chung, Duck-Hwa
    • Journal of Environmental Health Sciences
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    • v.36 no.1
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    • pp.52-60
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    • 2010
  • High performance liquid chromatography (HPLC) and liquid chromatography mass spectrometry (LC/MS) have been widely used to quantify aflatoxins in food, but these methods are expensive, time-consuming, unsuitable for analysis of the routine screening of large sample numbers and require derivatization and high level techniques to perform. The objective of this study is to detect aflatoxins in a large number of foods by a high efficient analytical system of combined enzyme linked immunosorbent assay (ELISA) for screening and LC/MS/MS for confirmation. The samples spiked individually with aflatoxin $B_1$ (0.5 and 1.0 ng/g) and total aflatoxins (10 ng/g) were analyzed by ELISA and LC/MS/ MS, and the recoveries for ELISA and LC/MS/MS were 71.8~119.2% and 70.8~135.3%, respectively. A total of 378 samples (grains, nuts, soybean and fermented soybean foods, pepper and fermented pepper foods) were purchased from the six major cities in Korea and analyzed by ELISA-LC/MS/MS system. Twenty two (5.8%; peanut: 11, pistachio: 2, walnut: 6, almond: 1, pepper powder: 1, pepper paste: 1) out of 378 samples were screened as aflatoxin B1 positive by ELISA, but, 4 (1.1%; peanut: 2, pistachio:1, pepper powder: 1) out of the 22 samples screened were confirmed as aflatoxins positive at levels of 1.02~52.79 ng/g by LC/MS/MS. ELISA-LC/MS/MS system provides a more rapid, accurate and cost-effective method for the detection of aflatoxins in large number of samples.

Effect of Coexisting Ions on Electrokinetic Injection in Capillary Electrophoresis Analysis (모세관 전기영동 분석에서 계면 동전기 주입에 미치는 공존 이온의 영향)

  • Lee, Kwang-Woo;Jeon, Ji-Young;Lee, Kwang-Pill
    • Analytical Science and Technology
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    • v.9 no.1
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    • pp.35-42
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    • 1996
  • A rapid analytical method based on capillary electrophoresis is described for the determination of trace anions in high-purity chemicals which is used to prevent corrosion demage in nuclear power plants. Separations are carried out at 20kV using trimethylsilane-coated fused-silica capillary ($70cm{\times}50$ or $75{\mu}m$ i.d.) with the electrolyte of 5mM Chromate(pH=8). Detection was achieved using on-column indirect photometry at 254nm. The simultaneous analysis of inorganic anions, chloride, nitrate, sulfate, azide and phosphate was performed using methods of hydrodynamic(>1ppm) and / or electrokinetic(<1ppm) injection. The results of studies on the coexisting anions on analyte ions shows that peak responses of analyte in hydrodynamic injection is constant without effect of coexisting anions, but those of analysis in electrokinetic injection is strongly dependant upon the kind of coexisting anions and its ionic mobility. The analyte enrichment rate, hence peak response, is positive relationship with the resistance of the sample solution. Thus, appropriate measures, such as standard addition or internal standard technique, must be used to account for differences in conductance of standard and sample solutions.

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Use of 16S-23S rRNA Intergenic Spacer Region for Rapid Detection of Vibrio fluvialis (16S-23S rRNA Intergenic Spacer Region을 이용한 Vibrio fluvialis의 검출)

  • 강현실;허문수;이제희
    • Korean Journal of Environmental Biology
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    • v.21 no.1
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    • pp.77-85
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    • 2003
  • We have examined the 16S-23S rRNA intergenic spacer regions (ISR) of Vibrio fluvialis. ISRs were PCR amplified, cloned into a plasmid vector and then sequenced. As results of ISR nucleotide sequence analysis, total of 6 clones were isolated depending on the size. The clones were different in both the number and the composition of the tRNA genes, and were designated ISR-A, ISR-E, ISR-El, ISR-lA, ISR-EKV, ISR-EKAV. ISR-A contains $tRNA^{Ala}$; ISR-lA, $tRNA^{Ile}$-$tRNA^{Ala}$; ISR-EKV, $tRNA^{GIu}$-$tRNA^{Lys}$-$tRNA^{Val}$;ISE-EKAV, $tRNA^{GIu}$-$tRNA^{Lys}$-$tRNA^{Ala}$-$tRNA^{Val}$; ISR -E and E1, $tRNA^{GIu}$ clusters. ISR-EKV was shown to be a minor type out of the six ISR types and showed a very limited homology between ISR-EKV from V, fluvialis and ISRa from other Vibrio species. Therefore ISR-EKV sequence was used to design species-specific primers to detect V, fiuvialis from other Vibrio species by PCR reaction. The specificity of the primers was examined using genomic DNA of other Vibrios as templates for PCR reaction. The result showed that PCR can be a useful method to detect V. fluvialis among Vibrio species in a single PCR reaction.

Sequence and Phylogenetic Analysis of Respiratory Syncytial Virus Isolated from Korea (국내에서 유행한 Respiratory Syncytial 바이러스의 염기서열 및 계통분석)

  • Kwon, Soon-Young;Choi, Young-Ju;Kim, So-Youn;Song, Ki-Joon;Lee, Yong-Ju;Choi, Jong-Ouck;Seong, In-Wha
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.9-22
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    • 1996
  • Respiratory Syncytial virus (RSV) is an important cause of acute lower respiratory tract infections in human, with infants and young children being particularly susceptible. In the temperate zones, sharp annual outbreaks of RSV occur during the colder months, in both the northern and the southern hemisphere. RSV is unusual in that it can repeatedly reinfect individuals throughout life and infect babies in the presence of maternal antibody. RSV isolates can be divided into two subgroups, A and B, on the basis of their reactions with monoclonal antibodies, and the two subgroups are also distinct at the nucleotide sequence level. The specific diagnosis of RSV infection was best made by isolation of virus in tissue culture, identification of viral antigen, or by specific serologic procedures. Recently, rapid detection of RSV and analysis of RSV strain variation became possible by development of methods of reverse transcription and polymerase chain reaction amplification. In this study, to determine the genetic diversity of RSV found in Korea, 173 bp and 164 bp spanning selected regions of the RSV F and SH genes were enzymatically amplified and sequenced, respectively. Eight for F gene and three for SH gene were detected in 66 nasopharyngeal swap samples tested. Two major antigenic subgroups, A and B were confirmed from Korean samples (seven for subgroup A and one for subgroup B). At the nucleotide level of the F gene region, Korean subgroup A strains showed 95-99% homologies compared to the prototype A2 strain of subgroup A and 93-100% homologies among Korean subgroup A themselves. For the SH gene region, Korean subgroup A strain showed 97.5% homology compared to the prototype A2 strain of subgroup A, and Korean subgroup B strain showed 97% homology compared to the prototype 18537 strain of subgroup B. Most of base changes were transition and occured in codon position 3, which resulted in amino acid conservation. Using the maximum parsimony method, phylogenetic analysis indicated that Korean RSV strains formed a group with other RSV strains isolated from the United States, Canada, the Great Britain and Australia.

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A study on the Private Investigator usage for Cyber Crime (사이버범죄의 효과적인 대응을 위한 민간조사제도의 도입방안)

  • Shin, Hyun-Joo
    • Korean Security Journal
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    • no.46
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    • pp.63-86
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    • 2016
  • The object of this study is to propose a study on the Private Investigator usage for Cyber Crime. The latest trend of cyber crime is being evolve in sophisticated and complex way over the global, like internet fraud, cyber gambling, hacking and etc. Hence national investigative authority mobilize high specialized skills and method of criminal investigation by each nation. But it is hard to respond in rapid and effective way because of propoor, distribution of group and insufficient of related legal system. Already in other countries, not considerable amount of services are given to private investigators in detection and tracking part which is inefficient by nation. So it has significantly meaningful to compensate the defect and study about private investigator usage as companion of cooperation policing for effectively respond to cyber-crime. The way to effectively deal with the cyber-crime is reevaluate meaning of partnership policing and need of private investigator usage. Also it is to analyze the main issue about introduction of a system and suggest the effective way of introduction. First, legislation of private investigator usage which is based upon partnership policing should be made up. Moreover, to establish the range of private investigator's business and enhance the reliability, it is to propose introduction of leading professional global certificate and license system with sufficient education and test. We are expecting introduction of private investigator usage can improve efficiency of investigation and promote effective countermeasures of cyber-crime.

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Development of Silm Type ELCB For Airport Distribution Panel through Increased short Circuit Capacity (단락용량 증대를 통한 슬림형 공항 분전반용 누전 차단기 개발)

  • Joo, Nam-Kyu;Lee, Jong-Myong;Kim, Nam-Ho
    • Journal of Advanced Navigation Technology
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    • v.16 no.2
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    • pp.360-366
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    • 2012
  • In the power distribution panel installed in airport or industrial facilities, MCCB has been used for main switch and ELCB for branch switch to perform human body and leakage-inducing fire protection as well as overcurrent and short circuit protection. Especially for the airport panel, increase in accident protection is needed for stable power supply due to rapid modernization with fast-growing users, higer capacity and diversification of equipment, the increase of power capacity and the breaker made slim is a main issue for now because the issue for installation space is standing out by making panel with two-row arrangement connection method, etc. due to a many use of branch ELCBs. In this thesis, we designed arc extinguishing mechanism, considered movement direction change of contact in mechanism design. Also, we designed the breaker to work stably in case of miniaturization of leakage detection circuit and reverse connection. We conducted short circuit test to verify its function and developed the breaker that can be improved protection against accidental current with slim size operating leakage function when reverse connection to help solve the problem in using space that is increasing in the airport distribution panel.

The Characteristics analysis of a Flux-lock Type Fault Current Limiter according to the Winding Directions for Power Grid (전력계통 적용을 위한 결선방향에 따른 자속구속형 한류기의 특성 분석)

  • Lee, Mi-Yong;Park, Jeong-Min
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.11
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    • pp.5879-5884
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    • 2013
  • With the rapid industrialization and economical development, the electricity demands of the industrial facilities and densely populated large cities are continuing to increase in Korea. The increase in the power consumption requires the extension of power facilities, but it is difficult to secure spaces for equipment installation in the limited space of urban areas. In addition, the 154 kV or 345 kV transmission systems in Korea has a short transmission distance, and are connected to one another in network structures that ensure the high reliability and stability of power supply. This structure reduces the impedance during the fault in power system, and increases the magnitude of in the short circuit fault current. The superconducting fault current limiter (SFCL) was devised to effectively address these existing problems. The SFCL is a new-concept eco-friendly protective device that ensures fast operation and recovery time for the fault current and does not need additional fault detection devices. Therefore, many studies are being conducted around the world. In this paper, based on the wiring method the initial fault current characteristics, current limiting characteristics, according to the incident angle and the change in inductance current limiting characteristics were analyzed in a multifaceted methods.

Meat Species Identification using Loop-mediated Isothermal Amplification Assay Targeting Species-specific Mitochondrial DNA

  • Cho, Ae-Ri;Dong, Hee-Jin;Cho, Seongbeom
    • Food Science of Animal Resources
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    • v.34 no.6
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    • pp.799-807
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    • 2014
  • Meat source fraud and adulteration scandals have led to consumer demands for accurate meat identification methods. Nucleotide amplification assays have been proposed as an alternative method to protein-based assays for meat identification. In this study, we designed Loop-mediated isothermal amplification (LAMP) assays targeting species-specific mitochondrial DNA to identify and discriminate eight meat species; cattle, pig, horse, goat, sheep, chicken, duck, and turkey. The LAMP primer sets were designed and the target genes were discriminated according to their unique annealing temperature generated by annealing curve analysis. Their unique annealing temperatures were found to be $85.56{\pm}0.07^{\circ}C$ for cattle, $84.96{\pm}0.08^{\circ}C$ for pig, and $85.99{\pm}0.05^{\circ}C$ for horse in the BSE-LAMP set (Bos taurus, Sus scrofa domesticus and Equus caballus); $84.91{\pm}0.11^{\circ}C$ for goat and $83.90{\pm}0.11^{\circ}C$ for sheep in the CO-LAMP set (Capra hircus and Ovis aries); and $86.31{\pm}0.23^{\circ}C$ for chicken, $88.66{\pm}0.12^{\circ}C$ for duck, and $84.49{\pm}0.08^{\circ}C$ for turkey in the GAM-LAMP set (Gallus gallus, Anas platyrhynchos and Meleagris gallopavo). No cross-reactivity was observed in each set. The limits of detection (LODs) of the LAMP assays in raw and cooked meat were determined from $10pg/{\mu}L$ to $100fg/{\mu}L$ levels, and LODs in raw and cooked meat admixtures were determined from 0.01% to 0.0001% levels. The assays were performed within 30 min and showed greater sensitivity than that of the PCR assays. These novel LAMP assays provide a simple, rapid, accurate, and sensitive technology for discrimination of eight meat species.