• 제목/요약/키워드: real-time polymerase chain reaction

검색결과 828건 처리시간 0.035초

Quantitative Real-Time PCR Assay for Detection of Paenibacillus polymyxa Using Membrane-Fusion Protein-Based Primers

  • Cho, Min Seok;Park, Dong Suk;Lee, Jung Won;Chi, Hee Youn;Sohn, Soo-In;Jeon, Bong-Kyun;Ma, Jong-Beom
    • Journal of Microbiology and Biotechnology
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    • 제22권11호
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    • pp.1575-1579
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    • 2012
  • Paenibacillus polymyxa is known to be a plant-growth-promoting rhizobacterium. The present study describes a quantitative polymerase chain reaction (qPCR) assay for the specific detection and quantitation of P. polymyxa using a primer pair based on the sequence of a membrane-fusion protein for the amplification of a 268 bp DNA fragment. This study reports that the qPCR-based method is applicable for the rapid and sensitive detection of P. polymyxa and can be used as an alternative method for agricultural soil monitoring.

Microarray Analysis of Gene Expression by Ginseng Water Extracts in a Mouse Adrenal Cortex after Immobilization Stress

  • Kim, Young-Ock;Lee, Sang-Won
    • Journal of Ginseng Research
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    • 제35권1호
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    • pp.111-123
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    • 2011
  • To investigate the effects of repeated immobilization-stress challenge on the the hypothalamic-pituitary-adrenal axis, the genomic transcriptome in the adrenal cortex of immobilization-stressed mouse was analyzed by using a cDNA microarray. Mice were subjected to immobilization stress for 2 h per day for 5 consecutive d. With a 4.0-fold cutoff of arbitrary criteria, the expression levels of 168 out of 41,174 genes were significantly modulated in the adrenal cortex by stress when comparing the control and experimental groups. These genes were related to apoptosis, cell cycle, immune response, inflammatory responses, and signal transduction, and thus may be used as potential targets for the development of therapeutics for chronic stress or depression. Six significant genes among these were selected for real time polymerase chain reaction analysis to confirm the change of their expression levels. The gene for phospho 1 was also further investigated because its expression showed the greatest fold-change.

이종 조직에서 사람 유전자 검출을 위한 디지털 중합효소연쇄반응의 적용 (Application of Digital Polymerase Chain Reaction for Human Gene Detection in Heterologous Tissues)

  • 김진희
    • 한국콘텐츠학회:학술대회논문집
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    • 한국콘텐츠학회 2018년도 춘계 종합학술대회 논문집
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    • pp.561-562
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    • 2018
  • 디지털 중합효소연쇄반응(Digital PCR)은 3세대 PCR로 명명하며, 1세대인 일반 PCR과 2세대인 정량 PCR(Real-time PCR)의 단점을 보완하여 개발된 방법이다. Digital PCR System은 소량의 PCR 반응을 10만개 이상의 반응통(wall)에 적재하는 방식의 나노유체칩에서 쪼개어 증폭시킨 후, target DNA를 계수한다. Target DNA의 증폭 여부에 따라 positive(1)와 negative(0)로 digital signal처럼 받아들여 계수하고, 포아송 분포를 통해 target DNA의 copy를 계산해 최종적으로 샘플 microlitr당 Copy수로 결과 값을 확인할 수 있다. 본 연구에서는 종(種)이 다른 동물의 조직이 서로 섞여있을 때 사람의 조직을 탐색하는 방법으로 유전자 증폭을 할 경우, digital PCR의 유효성에 대해 증명하였다.

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Molecular detection of bat coronaviruses in three bat species in Indonesia

  • Dharmayanti, Ni Luh Putu Indi;Nurjanah, Diana;Nuradji, Harimurti;Maryanto, Ibnu;Exploitasia, Indra;Indriani, Risa
    • Journal of Veterinary Science
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    • 제22권6호
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    • pp.70.1-70.12
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    • 2021
  • Bats are an important reservoir of several zoonotic diseases. However, the circulation of bat coronaviruses (BatCoV) in live animal markets in Indonesia has not been reported. Genetic characterization of BatCoV was performed by sequencing partial RdRp genes. Real-time polymerase chain reaction based on nucleocapsid protein (N) gene and Enzyme-linked immunosorbent assay against the N protein were conducted to detect the presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral RNA and antibody, respectively. We identified the presence of BatCoV on Cynopterus brachyotis, Macroglossus minimus, and Rousettus amplexicaudatus. The results showed that the BatCoV included in this study are from an unclassified coronavirus group. Notably, SARS-CoV-2 viral RNA and antibodies were not detected in the sampled bats.

Gene expression profiling after ochratoxin A treatment in small intestinal epithelial cells from pigs

  • Jung Woong, Yoon;Sang In, Lee
    • Journal of Animal Science and Technology
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    • 제64권5호
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    • pp.842-853
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    • 2022
  • Ochratoxin A (OTA) is a well-known mycotoxin that causes disease through the ingestion of contaminated food or feed, for example, in the porcine industry. The intestinal epithelium acts as the first barrier against food contamination. We conducted a study on the exposure of the porcine intestinal epithelium to OTA. We used the intestinal porcine epithelial cell line IPEC-J2 as an in vitro model to evaluate the altered molecular mechanisms following OTA exposure. Gene expression profiling revealed that OTA upregulated 782 genes and downregulated 896, totalling 1678 differentially expressed genes. Furthermore, immunofluorescence, quantitative real-time polymerase chain reaction, and western blotting confirmed that OTA damages the tight junction protein ZO-1. Moreover, OTA activated the expression of inflammatory genes (IL-6, IL-8, IL-10, NF-kB, TLR4, and TNF-α). In summary, this study confirmed that OTA alters various molecular mechanisms and has several adverse effects on IPEC-J2 cells.

Successful Management of Eosinophilic Bronchopneumopathy in a Dog

  • Kim, Sol;Seo, Kyoungwon;Song, Kunho
    • 한국임상수의학회지
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    • 제38권6호
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    • pp.269-273
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    • 2021
  • A dog was presented with a cough, dyspnea, nasal discharge, gagging, and exercise intolerance. The dog showed leukocytosis, peripheral eosinophilia, and an increase C-reactive protein. The radiographic findings noted bronchointerstitial infiltration, intrathoracic lymphadenopathy, and soft tissue opacity mass. Computed tomography findings showed thickening of the bronchus and bronchiole. Also, peri-bronchial consolidation and generalized intrathoracic lymphadenopathy was present. On blind bronchoalveolar lavage and pulmonary cytology, there were significantly increased eosinophils. Canine pulmonary respiratory pathogens from a real-time polymerase chain reaction analysis was negative. Consequently, the dog was diagnosed with eosinophilic bronchopneumopathy. Clinical signs improved significantly within a few days after treatment with an oral corticosteroid.

Anti-Inflammatory Effect of Carex scabrifolia Steud. Extract in RAW264.7 Cells

  • Joong Hyun Shim
    • 한국미생물·생명공학회지
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    • 제50권3호
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    • pp.354-360
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    • 2022
  • This research was designed to evaluate the possible anti-inflammatory effects of Carex scabrifolia Steud. extract using RAW264.7 cells. The assessments of these effects were based on cell viability assay, mRNA expression levels of interleukin-1 alpha (IL-1α), interleukin-1 beta (IL-1β), IL-6, tumor necrosis factor alpha (TNFα), and levels of nitric oxide (NO)/prostaglandin E2 (PGE2) production. Quantitative real-time polymerase chain reaction showed that treatment with C. scabrifolia Steud. extract decreased the mRNA levels of iNOS, COX2, IL-1α, IL-1β, IL-6, and TNFα. Furthermore, from the production levels of PGE2/NO, it can be inferred that C. scabrifolia Steud. extract exhibited anti-inflammatory properties. These results suggest that C. scabrifolia Steud. extract contains anti-inflammatory compound(s), and consequently, that it may have applications as a potent cosmeceutical material.

공기 중 박테리아 포집을 위한 습식 사이클론의 CFD 해석을 이용한 설계 및 성능 평가 (Design and Performance Evaluation using Computational Fluid Dynamics (CFD) Analysis of Wetcyclones for the Collection of Airborne Bacteria)

  • 고현식;박정우;정지우;황정호
    • 한국입자에어로졸학회지
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    • 제19권3호
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    • pp.77-87
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    • 2023
  • We present the development of a wetcyclone sampler designed for the sampling of airborne bacteria. The wetcyclone design involves a combination of two traditional cyclone shapes and computational fluid dynamics (CFD) analysis to validate its effectiveness in terms of pressure drop and collection efficiency. The wetcyclone exhibits a collection efficiency of over 90% for bacteria, specifically targeting Staphylococcus aureus. Additionally, the wetcyclone enables continuous bioaerosol sampling using a liquid medium (deionized water), demonstrating a concentration ratio exceeding >105 and a stable microbial recovery rate of 81.9%. The application of real-time quantitative polymerase chain reaction (qPCR) and the colony counting method ensures precise measurement of the concentration ratio and microbial recovery rate.

2019 Novel Coronavirus (COVID-19) Pneumonia: Serial Computed Tomography Findings

  • Jiangping Wei;Huaxiang Xu;Jingliang Xiong;Qinglin Shen;Bing Fan;Chenglong Ye;Wentao Dong;Fangfang Hu
    • Korean Journal of Radiology
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    • 제21권4호
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    • pp.501-504
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    • 2020
  • From December 2019, Coronavirus disease 2019 (COVID-19) pneumonia (formerly known as the 2019 novel Coronavirus [2019-nCoV]) broke out in Wuhan, China. In this study, we present serial CT findings in a 40-year-old female patient with COVID-19 pneumonia who presented with the symptoms of fever, chest tightness, and fatigue. She was diagnosed with COVID-19 infection confirmed by real-time reverse-transcriptase-polymerase chain reaction. CT showed rapidly progressing peripheral consolidations and ground-glass opacities in both lungs. After treatment, the lesions were shown to be almost absorbed leaving the fibrous lesions.

Identification and Analysis of the Chloroplast rpoC1 Gene Differentially Expressed in Wild Ginseng

  • Lee, Kwang-Ho;Kwon, Ki-Rok;Kang, Won-Mo;Jeon, Eun-Mi;Jang, Jun-Hyeog
    • 대한약침학회지
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    • 제15권2호
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    • pp.20-23
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    • 2012
  • Panax ginseng is a well-known herbal medicine in traditional Asian medicine, and wild ginseng is widely accepted to be more active than cultivated ginseng in chemoprevention. However, little has actually been reported on the difference between wild ginseng and cultivated ginseng. Thus, to identify and analyze those differences, we used suppressive subtraction hybridization (SSH) sequences with microarrays, realtime polymerase chain reaction (PCR), and reverse transcription PCRs (RT-PCRs). One of the clones isolated in this research was the chloroplast rpoC1 gene, a ${\beta}$subunit of RNA polymerase. Real-time RT-PCR results showed that the expression of the rpoC1 gene was significantly upregulated in wild ginseng as compared to cultivated ginseng, so, we conclude that the rpoC1 gene may be one of the important markers of wild ginseng.