• Title/Summary/Keyword: rapid detection method

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Detection of Apple Scar Skin Viroid by Reverse Transcription Recombinase Polymerase Amplification Assay

  • Kim, Na-Kyeong;Lee, Hyo-Jeong;Ryu, Tae-Ho;Cho, In-Sook;Ju, Ho-Jong;Jeong, Rae-Dong
    • Research in Plant Disease
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    • v.27 no.2
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    • pp.79-83
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    • 2021
  • The aim of the present study was to develop a sensitive and specific detection method for the rapid detection of apple scar skin viroid (ASSVd) in apple leaves. The resulting reverse transcription recombinase polymerase amplification (RT-RPA) assay can be completed in 10 min at 42℃, is 10 times more sensitive than conventional reverse transcription polymerase chain reaction, and can specifically amplify ASSVd without any cross-reactivity with other common apple viruses, including apple stem grooving virus, apple stem pitting virus, and apple chlorotic leaf spot virus. The reliability of the RT-RPA assay was assessed, and the findings suggested that it can be successfully utilized to detect ASSVd in field-collected samples. The RT-RPA assay developed in the present study provides a potentially valuable means for improving the detection of ASSVd in viroid-free certification programs, especially in resource-limited conditions.

Use of RAPD-PCR(Random Amplified Polymorphic DNA-Polymerase Chain Reaction) Method for a Detection of Pathogenic Listeria monocytogenes (RAPD-PCR(Random Amplified Polymorphic DNA - Polymerase Chain Reaction) 방법을 이용한 Listeria monocytogenes의 검색)

  • Park Bum-Joon;Sihn Eon-Hwan
    • The Korean Journal of Food And Nutrition
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    • v.17 no.3
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    • pp.254-259
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    • 2004
  • Rapid detection of foodbome pathogens is becoming increasingly important. The requirement for faster, more reliable tests has lead to the development of a wide range of rapid methods. Among these methods, the use of systems based on nucleic acid based detection has been increasing since they offer advantages of reduction in test time and more reliable detection or identification. Random Amplification Polymorphic DNA(RAPD) method has been used to fingerprint foodbome microorganisms; Listeria monocytogenes. In this study, 10-mer primer OPG-13(5'-CTCTCCGCCA-3') was used to generate RAPD-PCR for detection of pathogenic L. monocytogenes of Listeria spp. Among 20 primers tested, OPG-13 showed on acceptable result for the differentiation of a pathogenic Listeria from non-pathogenic microorganisms. Pathogenic Listeria, L. monocytogenes(ATCC 15313, 19111, 19112, 19113) showed two bands for 700 bp and 1,500 bp while non-pathogenic bacteria, L. ivanovii, L. grayi, L. murrayi, L. innocua, L. welshimeri, and L. seeligeri had only one band sizing from 2,000 to 2,300 bp. This RAPD method proved to be a valuable to gain important information on sources of pathogenic bacteria in food industry.

The Rapid Detection of Pathogens in Organically Grown Vegetables Using PCR-DGGE (PCR-DGGE를 이용한 유기농 채소의 유해 미생물 신속 검지)

  • Kwon, Oh Yeoun;Son, Seok Min
    • Food Engineering Progress
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    • v.15 no.4
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    • pp.370-375
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    • 2011
  • In this study the polymerase chain reaction (PCR) combined with denaturing gradient gel electrophoresis (DGGE) was evaluated as a method permitting the rapid detection of pathogens in fresh originally grown vegetables. A universal primer (341GCf/534r) was selected for its ability to amplify the V3 region of 16S-rRNA genes in their target pathogens (Salmonella typhimurium, Pseudomonas fluorescens, Bacillus cereus, Listeria monoytogenes, Staphyloocus aureus, E. coli). The 194 bp fragments in PCR were successfully duplicated as expected. The amplified fragments of the same size from six different pathogens also showed good separation upon DGGE. The detection limit of PCR-DGGE for six pathogens in fresh-cut lettuces were over $10^{5}$ CFU/g when sampled by stomaching. However, when the sampling method was changed from stomaching to shaking, the detection limit of six pathogens in organic vegetables was shown to increase by over $10^{1}$ CFU/g, but only those of B. cereus were over $10^{3}$ CFU/g. Therefore, PCR-DGGE was shown to be a reliable method for the detection of pathogens in fresh-cut vegetables.

Rapid Detection of Pathogens Associated with Dental Caries and Periodontitis by PCR Using a Modified DNA Extraction Method (PCR을 이용한 치아우식증 및 치주염 연관 병원체의 빠른 검출)

  • Kim, Jaehwan;Kim, Miah;Lee, Daewoo;Baik, Byeongju;Yang, Yeonmi;Kim, Jaegon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.41 no.4
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    • pp.292-297
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    • 2014
  • DNA extraction is a prerequisite for the identification of pathogens in clinical samples. Commercial DNA extraction kits generally involve time-consuming and laborious multi-step procedures. In the present study, our modified DNA isolation method for saliva samples allows for the quick detection of pathogens associated with dental caries or periodontitis by PCR within 1 h. To release DNA from the bacteria, 1 min of boiling was adequate, and the resulting isolated DNA can be used many times and is suitable for long term storage of at least 13 months at $4^{\circ}C$, and even longer at $-20^{\circ}C$. In conclusion, our modified DNA extraction method is simple, rapid, and cost-effective, and suitable for preparing DNA from clinical samples for PCR for the rapid detection of oral pathogens from saliva.

Performance Evaluation of a Rapid Three Dimensional Diffusion MRI

  • Numano, Tomokazu;Homma, Kazuhiro;Nishimura, Katsuyuki
    • Proceedings of the Korean Society of Medical Physics Conference
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    • 2002.09a
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    • pp.356-358
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    • 2002
  • MRI, particularly diffusion weighted imaging (DWI), plays vital roles in detection of the acute brain infarction$\^$1-4/ and others metabolic changes of biological tissues. In general, every molecule in biological tissues may diffuse and move randomly in three-dimensional space. However, in clinical diagnosis, only 2D-DWI is used. The authors have developed a new method for rapid three-dimensional DWI (3D-DWI). In this method, by refocusing of the magnetized spin with the applied gradient field, direction of which is opposite to phase encoding field. Magnetized spin of $^1$H is kept under the SSFP (steady state free precession)$\^$5-6/. Under SSFP, in addition of FID, spin echo and stimulated echo are also generated, so the acquired signal is increased. The signal intensity is increased depending on flip angle (FA) of magnetized spin. This phenomenon is confirmed by human brain and phantom studies. The performance of this method is quantitatively analyzed by using both of conventional spin echo DWI and 3D-DWI. From experimental results, three dimensional diffusion weighted images are obtained correctly for liquid phantoms (water, acetone and oil), diffusion coefficient is enhanced in each image. Therefore, this method will provide useful information for clinical diagnosis.

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Determination of Cholesterol in Milk and Dairy Products by High-Performance Liquid Chromatography

  • Oh, H.I.;Shin, T.S.;Chang, E.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.10
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    • pp.1465-1469
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    • 2001
  • A sensitive high-performance liquid chromatographic method was developed to determine the content of cholesterol in milk and dairy products. To optimize separation of cholesterol, mobile phases including acetonitrile:2-propanol (8:1, v/v), acetonitrile:methanol (3:1, v/v), and acetonitrile:methanoI:2-propanol (7:3: I, v/v/v) were compared. Acetonitrile/methanol/2-propanol was superior to the other mobile phase systems for separating cholesterol. Liquid-liquid extraction (LLE) of cholesterol was simplified using a non-polar solvent, hexane, to remove interfering compounds, and had an excellent recovery $(100{\pm}1.0%)$ of cholesterol. A solid phase extraction (SPE) method using Sep-pak $C_{18}$ was developed and compared with LLE. The SPE method was rapid and highly reproducible. Both extraction methods were useful when used in combination with saponification of esterified cholesterol to facilitate total cholesterol determination. The detection limit of cholesterol was $0.01{\mu}g$. The newly developed HPLC method was rapid, simple, and accurate, and has advantages over the many methods commonly used.

Effective Error Detection Method for Video using Fragile Watermark (연성 워터마크를 이용한 비디오의 효율적인 에러 검출 방법)

  • Hwang, Young-Hooi;Jeon, Byeung-Woo
    • Journal of the Institute of Electronics Engineers of Korea SP
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    • v.39 no.6
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    • pp.602-611
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    • 2002
  • Recent advances in technology have resulted in rapid growth in mobile communication. With this explosive growth, reliable transmission and error resilient technique become increasingly necessary to offer high quality multimedia service. The success of error resilient techniques at decoder sensitively depends on error detection performance. Therefore, this paper proposes a computationally very simple and efficient error detection technique using fragile watermark for real-time video communication. To balance between image quality degradation and error detection efficiency, fragile watermark is embedded only in least significant bits of selected transform coefficients. The proposed method is workable without additional bit in video bitstream and can be implemented very efficiently. This method will be useful in video communication in error prone environment such as wireless channel.

Highly Sensitive Biological Analysis Using Optical Microfluidic Sensor

  • Lee, Sang-Yeop;Chen, Ling-Xin;Choo, Jae-Bum;Lee, Eun-Kyu;Lee, Sang-Hoon
    • Journal of the Optical Society of Korea
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    • v.10 no.3
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    • pp.130-142
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    • 2006
  • Lab-on-a-chip technology is attracting great interest because the miniaturization of reaction systems offers practical advantages over classical bench-top chemical systems. Rapid mixing of the fluids flowing through a microchannel is very important for various applications of microfluidic systems. In addition, highly sensitive on-chip detection techniques are essential for the in situ monitoring of chemical reactions because the detection volume in a channel is extremely small. Recently, a confocal surface enhanced Raman spectroscopic (SERS) technique, for the highly sensitive biological analysis in a microfluidic sensor, has been developed in our research group. Here, a highly precise quantitative measurement can be obtained if continuous flow and homogeneous mixing condition between analytes and silver nano-colloids are maintained. Recently, we also reported a new analytical method of DNA hybridization involving a PDMS microfluidic sensor using fluorescence energy transfer (FRET). This method overcomes many of the drawbacks of microarray chips, such as long hybridization times and inconvenient immobilization procedures. In this paper, our recent applications of the confocal Raman/fluorescence microscopic technology to a highly sensitive lab-on-a-chip detection will be reviewed.

Feasibility Study for Detection of Turnip yellow mosaic virus (TYMV) Infection of Chinese Cabbage Plants Using Raman Spectroscopy

  • Kim, Saetbyeol;Lee, Sanguk;Chi, Hee-Youn;Kim, Mi-Kyeong;Kim, Jeong-Soo;Lee, Su-Heon;Chung, Hoeil
    • The Plant Pathology Journal
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    • v.29 no.1
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    • pp.105-109
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    • 2013
  • Raman spectroscopy provides many advantages compared to other common analytical techniques due to its ability of rapid and accurate identification of unknown specimens as well as simple sample preparation. Here, we described potential of Raman spectroscopic technique as an efficient and high throughput method to detect plants infected by economically important viruses. To enhance the detection sensitivity of Raman measurement, surface enhanced Raman scattering (SERS) was employed. Spectra of extracts from healthy and Turnip yellow mosaic virus (TYMV) infected Chinese cabbage leaves were collected by mixing with gold (Au) nanoparticles. Our result showed that TYMV infected plants could be discriminated from non-infected healthy plants, suggesting the current method described here would be an alternative potential tool to screen virus-infection of plants in fields although it needs more studies to generalize the technique.

A Detection Kit for Aeromonas hydrophila Using Antibody Sensitized Latex

  • Shin, En-Joo;Lee, Soon-Deuk;Lee, Kyung-Won;Lee, Yeon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.10 no.5
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    • pp.595-598
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    • 2000
  • Aeromonas hydrophila is a pathogen to fish as well as human. It is a food-borne disease, and causes severe mortality in fish, and sometimes severe septicemia in human. In this study, a rapid detection method using latex agglutination has been developed for A. hydrophila. Polyclonal antibodies were raised against membrane and whole cells of three isolates from rainbow trout. Among these, latex particles coated with antibodies raised against whole cells of isolate No. 2 showed the best sensitivity. With latex particles coated with this antibody, we could detect $5{\times}10^4$ CFU of A. hydrophila in 5 min. The cross-reactivity with bacteria constituting the normal intestinal microflora and other pathogens for rainbow trout was insignificant. This latex agglutination assay method produced positive reaction with all clinical isolates of A. hydrophila which were identified by species-specific PCR for 16S rRNA in A. hydrophila.

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