• Title/Summary/Keyword: PCR-ELISA

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MALDI TOF MS for the identification of Salmonella spp. from swine (돼지유래 Salmonella속 균의 동정을 위한 MALDI TOF MS 활용)

  • Sohn, Jun Hyung;Jeon, Woo Jin;Lee, Young Mi;Kim, Seon Soo
    • Korean Journal of Veterinary Service
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    • v.39 no.4
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    • pp.247-251
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    • 2016
  • Salmonella is one of the most common bacteria that causes heavy losses in swine industry and major causative pathogen of food poisoning in public health. Various methods for the identification of Salmonella such as Gram staining, agglutination test, enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR) have been used. Several studies have demonstrated that Matrix Assisted Laser Desorption Ionization Time of Flight (MALDI TOF) Mass Spectrometry (MS) identification is an efficient and inexpensive method for the rapid and routine identification of isolated bacteria. In this study, MALDI TOF MS could provide rapid, accurate identification of Salmonella spp. from swine compared with end point PCR and real time PCR.

Development of PCR Primers to Detect Pseudomonas savastanoi pv. phaseolicola from the Bean Seeds (강낭콩 종자에서 Pseudomonas savastanoi pv. phaseolicola의 검출을 위한 PCR 프라이머의 개발)

  • Cho, Jung-Hee;Jeong, Min-Jung;Song, Min-Ji;Yim, Kyu-Ock;Lee, Hyok-In;Kim, Jung-Hee;Baeg, Ji-Hyun;Cha, Jae-Soon
    • Research in Plant Disease
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    • v.16 no.2
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    • pp.129-135
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    • 2010
  • PCR primers were developed to detect Pseudomonas savastanoi pv. phaseolicola, a causal agent of halo blight that occurs in all species of common bean (Phaseolus vulgaris L.), from the bean seeds. A primer set, Psp-JHF and Psp-JH-R, specifically amplified 513 bp fragment from Pseudomonas savastanoi pv. phaseolicola only. A nested primer set, psp-JH-F-ne and psp-JH-R-ne, designed from the $1^{st}$ PCR amplicon, amplified 169 bp fragment. The primer sets did not amplify any non-specific DNA from the seed extracts of Fabaceae including 4 beans, 2 soybeans, and 2 peas. The detection sensitivity of the nested PCR method developed in this study was much higher than that of ELISA and selective medium. PCR assays developed in this study should be useful to detect Pseudomonas savastanoi pv. phasolicola from the bean seeds.

PCR에 의한 HIV의 진단

  • Kang, Chun
    • The Microorganisms and Industry
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    • v.18 no.2
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    • pp.26-30
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    • 1992
  • HIV는 ELISA나 WB에 의해 항체가 검출되기 수개월 혹은 수 년전에도 proviral DNA 상태로 감염된 세포의 chromosome내에 존재하는 것이 주지의 사실이다. 그동안 Southern blot, in situ hybridization등에 의해 이 proviral DNA를 검출하려는 연구가 진행되어 왔으나 lymphocyte $10^{4}$-$10^{6}$개 중 1개가 감염되어 있으며 lymphocyte chromosomal DNA에 비해 viral DNA의 양이 미량이므로 검출하기에는 민감도가 낮은 문제점이 있다. 본 고에서는 근래 개발되어 널리 사용되고 있는 polymerase chain reaction(PCR)을 이용한 HIV의 진단에 관해 살펴보고자 한다.

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Effect of Ferulic Acid Isolated from Cnidium Officinale on the Synthesis of Hyaluronic Acid (천궁으로부터 분리된 ferulic acid의 히알루론산 생성에 미치는 효과)

  • Song, Hye Jin;Jin, Mu Hyun;Lee, Sang Hwa
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.39 no.4
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    • pp.281-288
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    • 2013
  • Hyaluronic acid (HA) is one of the major extracellular matrix components in skin. The HA content is reported to decline with age, which may contribute to decrease in skin moisture, wrinkle formation and the decrease in elasticity of the skin. Among the family of HA synthase genes (HAS-1, 2, 3) identified so far, HAS-2 plays crucial roles in the regulation of HA synthesis in human skin fibroblasts. In this study, we elucidated the effects of ferulic acid isolated from Cnidium officinale on HA production. Semi-quantitative RT-PCR and quantitative real-time PCR showed that ferulic acid increased mRNA level of HAS-2 gene and ELISA assay also revealed that ferulic acid increased HA production in human skin fibroblasts. Our study suggests that ferulic acid might prevent age-dependent skin deteriorations such as wrinkles, dryness and elasticity decrease, all of which could be ascribed to the reduction of the HA content in human skin.

Evaluation of Invasive and Noninvasive Methods for the Diagnosis of Helicobacter Pylori Infection

  • Cosgun, Yasemin;Yildirim, Abdullah;Yucel, Mihriban;Karakoc, Ayse Esra;Koca, Gokhan;Gonultas, Alpaslan;Gursoy, Gul;Ustun, Huseyin;Korkmaz, Meliha
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.12
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    • pp.5265-5272
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    • 2016
  • Objective: The present study was conducted to evaluate invasive and noninvasive diagnostic methods for detection of Helicobacter pylori (H. pylori) in patients admitted with dyspeptic complaints and to compare sensitivities and specificities. Method: Sets of four gastric biopsy specimens were obtained from a total of 126 patients included in the study. The presence of H. pylori was determined by invasive tests including culture, rapid urease test, polymerase chain reaction (PCR) and histopathology. Among noninvasive tests, urea breath test, serological tests and enzyme-linked immunosorbent assay (ELISA) were performed. Results: H. pylori was isolated in 79 (62.7%) gastric biopsy cultures, whereas positivity was concluded for 105 (83.3%) patients by rapid urease test, for 106 (84.1%) by PCR, for 110 (87.3%) by histopathology, for 119 (94.4%) by urea breath test, and for 107 (84.9%) by ELISA. In the present study, the culture findings and histopathological examination findings were accepted as gold standard. According to the gold standard, urea breath test had the highest sensitivity (96.5%) and the lowest specificity (30%), whereas culture and histopathology had the highest specificities (100%). Conclusion: The use of PCR invasively with gastric biopsy samples yielded parallel results with the gold standard. PCR can be recommended for routine use in the diagnosis of H. pylori.

Identification of Introduced Gene and Its Expression and Gene Stability Assessment for Event Selection of Genetically Modified Plant toward Approval: Cucumber Mosaic Virus Resistant Hot Pepper (상업용 유전자 변형작물 이벤트 선발을 위한 도입유전자 확인, 발현 및 세대간 안정성 평가 : 오이모자이크바이러스 저항성 GM 고추)

  • Kang, Seung-Won;Han, Bal-Kum;Lee, Tae-Ho;Kim, Eun-Ji;Lee, Gung-Pyo
    • Horticultural Science & Technology
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    • v.30 no.2
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    • pp.192-200
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    • 2012
  • For the development of genetically modified plants, it is important to verify various factors which potentially affect the risk assessment as well as to establish an experimental program to produce scientific and reliable data. However, it is a time and cost consuming process to develop GM plants as well as to prepare scientific and convincible data for government's approval. Therefore, using the transgenic hot pepper tolerant to a new CMV pathotype, we attempted to suggest few methodological procedures, such as probe saturation for southern blot analysis and RT-PCR and ELISA for expression analysis, for identification and stability evaluation of inserted gene in genetically modified plant which are required for submission for approval. Ten partially overlapped probes covering full length of inserted gene were produced. We could identify that the inserted gene was stacked as a single copy as well as no partial element existed. Also, we could identify the stability of the inserted gene stacked in hot pepper using probe saturation. In the expression analysis with RT-PCR and ELISA, we also could provide the stable expression of transcript and proteins in leaves and placenta and pericarp of fruits of the CMV-resistant hot pepper.

Micrografting and Heat Treatment Combination for Eliminating Virus of CTV-infected Citrus (CTV 바이러스 보균 감귤나무로부터 열처리와 경정접목을 통한 바이러스 제거)

  • Chae, Chi Won;Yun, Su Hyun;Park, Jae Ho;Hyun, Jae Wook;Koh, Sang Wook;Lee, Dong Hoon
    • Journal of Life Science
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    • v.23 no.2
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    • pp.267-272
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    • 2013
  • This study was conducted to eliminate viruses from citrus-infected plants using micrografting and thermotherapy. Six citrus cultivars including a 'Setoka' hybrid were used as plant sources. The TAS-ELISA technique demonstrated that several plants were CTV positive. However, no CTV symptoms were detected in plants obtained from shoots and treated at a high temperature of $40^{\circ}C$ during the day and night and micrografted for two weeks with old trifoliate orange rootstock in vitro. Indexing of CTV, SDV, and CTLV for RT-PCR analysis of the eleven citrus seedlings, including 'Setoka', 'Samdajosang', 'Pungkwang', 'Shiranuhi', and 'Ehimekashi dai28go' was virus free following the micrografting and thermal therapy.

Simple and Rapid Identification of Low Level Hepatitis B Virus DNA by the Nested Polymerase Chain Reaction

  • Jang, Jeong-Su;Lee, Kong-Joo
    • Archives of Pharmacal Research
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    • v.19 no.6
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    • pp.469-474
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    • 1996
  • A rapid and sensitive method has been developed to detect hepatitis B virus DNA (HBV) by nested polymerase chain reaction (PCR) technique with primers specific for the surface and core regions in capillary thermal cycler within 80 min. The lower limit for detection by present PCR method is $10^{-5}$ pg of recombinant HBV DNA which is equivalent to that determined by one round of PCR amplification and Southern blot hybridization analysis. When boiled HBV positive serum was serially diluted 10-fold, HBV DNA was successfully determined in $1{\mu}l-10^{-3}$ of serum. HBV DNA was detected by present method in 69 clinical samples including HBsAg positives and negatives by enzyme-linked immunosorbent assay (ELISA). When serum samples were amplified by nested PCR using surface and core region primers, HBV DNAs were detected in 37 of 69 samples (53.6%) and 18 of 69 samples (26.1%), respectively. These results can inform the infectious state of HBsAg positive pateints. A simple and rapid nested PCR protocol by using boiled serum as DNA template has been described for the clinical utility to determine HBV DNA in human serum.

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Prevalence and Characterization of Enterohemorrhagic Escherichia coli (EHEC) Isolated from Ground Beefs Distributed in Gyeong-In Region (경인지역에 유통되는 분쇄육 중 장출혈성대장균의 분포 조사 및 특성 연구)

  • Kim, Eun-Jeong;Park, Yong-Chjun;Cho, Joon-Il;Lee, Jong-Ok;Kim, Hee-Yun
    • Korean Journal of Food Science and Technology
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    • v.38 no.6
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    • pp.773-778
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    • 2006
  • The objective of this study was to evaluate three verocytotoxin-producing Escherichia coli (VTEC) detection kits to detect the presence of VT genes: Doupath Verocytotoxin (GLISA) developed by MERCK, ProsPect Shiga Toxin E. coil (STEC) Microplate Assay (ELISA) developed by Remel, and a polymerase chain reaction method. Our laboratory verified artificially inoculated samples. All three methods could detect very low numbers of VTEC, but VT-PCR had the best sensitivity for VTEC detection. From April through September 2005, 257 ground-beefs from supermakets and traditional markets were examined for the presence of VTEC by polymerase chain reaction immediately after purchase and total viable counts (TVC) were determined. VTEC was isolated from 30 of 257 ground-beefs. A variety of serogroups was found, including 10 stains belonging to the virulence type EHEC, but major serogroups such as O157, O26 and O111 were nor found.

The prevalence of avian reovirus infection in poultry farms of Jeonbuk province, Korea (전북지역 양계농가의 조류레오바이러스 유병율 조사)

  • Jeong, Jae-Kyo;Jeong, Hansol;An, Euingyoub;Han, Seunghwan
    • Korean Journal of Veterinary Service
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    • v.42 no.4
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    • pp.237-243
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    • 2019
  • Avian reovirus (ARV) is the pathogenic agent of tenosynovitis and malabsorption syndrome in broiler, which has caused significant economical losses due to poor feeder efficiency and stunting. In order to determine the prevalence of ARV infection in poultry farms in Jeonbuk province, Korea, we performed a surveillance study by testing 179 cecal samples from 131 broiler farms for virus detection, and 1,181 serum samples from 33 broiler farms (n=292) and 22 broiler breeder farms (n=1,525) for antibody detection in the province. Virological examination using RT-PCR showed that ARV were detected in 26.0% of the tested farms (34/131),with the highest positive rates in broilers of 6 days old or more in summer season. In serological test using ELISA, broiler and broiler breeder farms examined were all ARV antibody positive. In broiler, the positive rate and antibody titers showed a tendency to decrease with age in contrast to those of broiler breeders. Our results indicate that ARV is ubiquitous in broilers and broiler breeders in the province.