• Title/Summary/Keyword: Strain specificity

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Isolation and Biological Characterization of Barley mild mosaic virus(BaMMV) Mild and Severe Strains in Korea

  • Jonson, Gilda;Park, Jong-Chul;Noh, Tae-Hwan;Kim, Mi-Jung;Hyun, Jong-Nae;Kim, Jung-Gon
    • The Plant Pathology Journal
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    • v.22 no.4
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    • pp.329-333
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    • 2006
  • Two distinct and stable isolates of Barley mild mosaic virus(BaMMV) designated as Naju82-S(severe) and Naju82-M(mild) were obtained. These two isolates differed in their symptomatology, virus transmission characteristics and cultivar specificity at various temperature. Thus, these isolates were referred to as strains in this study. BaMMV Naju-S strain showed severe mosaic symptoms accompanied by necrosis on the infected leaves. Naju82-S strain is more virulent demonstrated by shorter incubation period and relatively high virus concentration than Naju82-M strain. Five Korean cultivars were tested for their pathogenicity to different strains based on the rate of infection. Results showed that infection rate of cultivars to both strains did not significantly differed from each other. However, under different temperatures, the pathogenicity on the two cultivars such as cultivars Hopumbori and Sessalbori were significantly affected. Hopumbori was moderately resistant to both strains at $10-12^{\circ}C$ and susceptible at $15-18^{\circ}C$. Similarly, Sessalbori was moderately resistant at $10-12^{\circ}C$ to both strains but distinctly differentiated at $15-18^{\circ}C$ wherein it was resistant to mild strain and highly susceptible to severe strain. Other cultivars including Baegdong, Jinyangbori and Neahanssalbori consistently showed susceptible reaction to both strains at varying temperatures tested in this study.

Development of a Blocking ELISA for Measuring Rabies Virus-specific Antibodies in Animals

  • Yang, Dong-Kun;Kim, Ha-Hyun;Ryu, Jieun;Gee, Mi-ryun;Cho, In-Soo
    • Microbiology and Biotechnology Letters
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    • v.46 no.3
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    • pp.269-276
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    • 2018
  • Rabies virus (RABV)-specific antibodies in animals and humans are measured using standard methods such as fluorescent antibody virus neutralization (FAVN) tests and rapid fluorescent focus inhibition tests, which are based on cell culture systems. An alternative assay that is safe and easy to perform is required for rapid sero-surveillance following mass vaccination of animals. Two purified monoclonal antibodies (4G36 and B2H17) against RABV were selected as capture and detection antibodies, respectively. A genetically modified RABV, the ERAGS strain, was propagated and concentrated by polyethylene glycol precipitation. Optimal conditions for the RABV antigen, antibodies, and serum dilution for a blocking enzymelinked immune sorbent assay (B-ELISA) were established. We evaluated the sensitivity, specificity, and accuracy of the B-ELISA using serum samples from 138 dogs, 71 raccoon dogs, and 25 cats. The B-ELISA showed a diagnostic sensitivity of 95.8-96.3%, specificity of 91.3-100%, and accuracy of 96.0-97.2% compared to the FAVN test. These results suggest that the B-ELISA is useful for sero-surveillance of RABV in dogs, raccoon dogs, and cats.

Development of PCR-microplate Hybridization Assay for Detection of Mycobacterium tuberculosis

  • Lee, In-Soo;Cho, Een-Jin;Cho, Sang-Nae;Kim, Tae-Ue;Lee, Hye-Young
    • Biomedical Science Letters
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    • v.15 no.4
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    • pp.295-300
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    • 2009
  • Tuberculosis caused by Mycobacterium tuberculosis (MTB) still remains to be the most dreadful infectious disease affecting almost every country. In the present study, we developed a simple and rapid but accurate and sensitive assay method for detecting MTB using microplate hybridization assay. For this, a selective region of the rpoB gene was used to design PCR primers, and MTB and Mycobacterium genus-specific probe molecules. The specificity of the assay was confirmed using fifteen different mycobacterial reference strains and twelve different non-mycobacterial reference strains, and the sensitivity was determined to be 100 fg using genomic DNA of MTB reference strain, H37Rv. Subsequently, a total of 62 sputum samples with diverse smear scores and culture positive results were used to evaluate the kit performance. In brief, the specificity and the sensitivity of the assay were 100% and 98.4%, respectively.

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Neutralizing Chimeric Mouse-human Antibodies against Burkholderia pseudomallei Protease: Expression, Purification and Characterization

  • Chan, Shzu-Wei;Ong, Guan-Im;Nathan, Sheila
    • BMB Reports
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    • v.37 no.5
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    • pp.556-564
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    • 2004
  • A recombinant Fab monoclonal antibody (Fab) C37, previously obtained by phage display and biopanning of a random antibody fragment library against Burkholderia pseudomallei protease, was expressed in different strains of Escherichia coli. E. coli strain HB2151 was deemed a more suitable host for Fab expression than other E. coli strains when grown in media supplemented with 0.2% glycerol. The expressed Fab fragment was purified by affinity chromatography on a Protein G-Sepharose column, and the specificity of the recombinant Fab C37 towards B. pseudomallei protease was proven by Western blotting, enzyme-linked immunosorbent assay (ELISA) and by proteolytic activity neutralization. In addition, polyclonal antibodies against B. pseudomallei protease were produced in rabbits immunized with the protease. These were isolated from high titer serum by affinity chromatography on recombinant-Protein A-Sepharose. Purified polyclonal antibody specificity towards B. pseudomallei protease was proven by Western blotting and ELISA.

Rapid Identification of Candida albicans Using Colorimetric Method

  • Kim, Shin Young;Park, Hun-Hee
    • Korean Journal of Clinical Laboratory Science
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    • v.45 no.4
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    • pp.149-153
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    • 2013
  • Candidiasis is a fungal infection of the most common causes; generally, opportunistic infections occur often in patients with weakened immune systems. Because of high rates in fungal infection patients and increasing frequency of being isolated from clinical materials, quickly identifying of Candida albicans is critical. By identifying 404 yeast cell strains of referred samples via API 20C kits, NGL and PRO tests and Germ tube (GT) test were conducted and compared. In the 3.0 McFarland yeast cells, 0.1% ${\rho}-nitrophenyl-N-acetyl-{\beta}-D-galactosaminide$ (NGL) and 0.04% ${\small{L}}$-proline ${\beta}$-naphtylamide (PRO) were each put in test tubes and incubated at $35^{\circ}C$ for 15, 30, 60 and 90 minutes. Afterwards, 1 drop of 2% NaOH was applied, and if the color turned yellow; it was positive for NGL test. Afterwards, 1% ${\rho}$-dimethylaminocinnamaldehyde was applied, and if the upper layer turned pink or red, it was positive for PRO test. NGL and PRO tests were conducted for all C. albicans and identified accurately within 30 minutes. In NGL, PRO test, false-positive, negative were not seen, whereas, GT test showed false-positive in 1 strain and false-negative in 3 strains. Therefore, sensitivity and specificity of NGL, PRO tests were 100% and 99.5%, respectively, and positive and negative predictive rate were 99.5% and 100%, respectively. However, GT test sensitivity and specificity were 98.5% and 99.5%, respectively, and positive and negative predictive rates were 99.5% and 98.5%, respectively. In conclusion, NGL, PRO tests are better than GT tests for sensitivity and specificity, therefore, these reliable tests will be useful in clinical laboratories.

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Production and characterization of anti-Salmonella polyclonal antibodies as bio-recognition element for developing a microbial monitoring method (미생물학적 모니터링 분석방법 개발을 위한 생물학적 수용체로서 살모넬라에 특이적인 다중클론 항체의 생산 및 특성 검토)

  • Park, Mi-Kyung
    • Food Science and Preservation
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    • v.24 no.7
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    • pp.885-890
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    • 2017
  • For the construction of the microbial monitoring method, anti-Salmonella polyclonal antibodies (pAbs) were produced from a rabbit and purified by saturated ammonium sulfate precipitation and protein A affinity column. The reactivity of anti-Salmonella pAbs was compared to that of commercial ones by using an indirect ELISA. The specificity of anti-Salmonella pAbs was investigated using 20 Salmonella serotypes and 20 non-Salmonella strains. A capturing ability of anti-Salmonella pAbs was investigated by exposing antibody-immobilized gold biosensor to different concentration of Salmonella mixture. Anti-Salmonella pAbs were successfully produced and purified with an antibody concentration of 2.0 mg/mL The reactivity of purified anti-Salmonella pAbs was greater than that of commercial one at all tested concentrations. All Salmonella serotypes, except S. Diarizonae, showed excellent binding efficiency with purified anti-Salmonella pAbs. Moreover, the purified anti-Salmonella pAbs showed excellent specificity against all non-Salmonella strains. The anti-Salmonella pAbs immobilized on the gold biosensor demonstrated the successful capturing capability against Salmonella with a dose-response manner. Therefore, the anti-Salmonella pAbs exhibited sufficient reactivity, specificity, as well as capturing capability against Salmonella to be considered as a bio-recognition element.

Current Status of Photobiological Hydrogen Production Technology Using Unicellular Marine Cyanobacterial Strains (단세포성 해양남세균 종주를 이용한 광생물학적 수소생산 기술)

  • Park, Jong-Woo;Kim, Jae-Man;Yih, Won-Ho
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.14 no.1
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    • pp.63-68
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    • 2009
  • Among various microscopic organisms producing photobiological hydrogen, cyanobacteria have long been recognized as the promising biological agents for hydrogen economy in 21 century. For photobiological production of hydrogen energy, marine unicellular $N_2$-fixing cyanobacteria have been evaluated as an ideal subgroup of Cyanophyceae. To develope the hydrogen production technology using unicellular $N_2$-fixing cyanobacteria, 3 important factors are pre-requisite: 1) isolation of the best strain from marine natural environment, 2) exploration on the strain-specific optimal conditions for the photobiological hydrogen production, and finally 3) application of the molecular genetic tools to improve the natural ability of the strain to produce hydrogen. Here we reviewed the recent research & development to commercialize photobiological hydrogen production technology, and suggest that intensive R&D during next 10-15 years should be imperative for the future Korean initiatives in the field of the photobiological hydrogen production technology using photosynthetic marine unicellular cyanobacterial strains.

Production and characterization of monoclonal antibodies against porcine transmissible gastroenteritis virus (돼지 전염성 위장염 바이러스에 대한 단크론항체 생산 및 특성)

  • Jang, Young-en;Cho, Sun-hee;Kim, Byung-han;Ahn, Jae-moon;Kang, Shien-young
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.336-344
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    • 1998
  • Eight monoclonal antibodies(MAbs) against the transmissible gastroenteritis virus (TGEV) were produced and characterized. Four of the MAbs were produced against a reference TGEV, Purdue strain(P115) and the others were produced against the Korean vaccine virus, Pyungtaek strain. Only one MAb(5C8) produced against P115 had neutralizing activity and was found to be E2 protein-specific. The other seven MAbs(4E2, 4G8, 5H6, 1F8, 2C6, 5H5, and 3A6) had specificity of nucleocapsid protein and no neutralizing activity. All MAbs reacted with different strains of TGEV, but none of the MAbs was reactive with porcine enteropathogenic viruses such as rotavirus, epidemic diarrhea virus and enterovirus by fluorescence antibody(FA) test.

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Synchronization of Cell Cycle in Korean Hydrogen Producing Cyanobacterial Strains (한국산 수소생산 남세균 종주들의 세포주기 동조화)

  • Park, Jong-Woo;Ahn, Se-Hee;Kim, Hyung-Seop;Yih, Won-Ho
    • Transactions of the Korean hydrogen and new energy society
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    • v.22 no.5
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    • pp.663-670
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    • 2011
  • Under a daily photoperiod of 14h light and 10h dark synchronization of cell cycle in Korean Cyanothece spp. strains and $Synechococcus$ sp. strain Miami BG043511 was analyzed as to be applicable to enhanced hydrogen production. For all strains peaks of double cell were observed during the light period of a daily cycle. Peaks of maximal cell size measured by a coulter counter appeared at the peak of double cells observed under light microscope reconfirming the synchronization of daily cell cycle. The cell cycle synchronization became weakened within two days when treated with continuous illumination. Rapid detection of the peak time of double cell percentage by coulter counters may contribute to quasi-realtime feedback control for efficient production of photobiological hydrogen by unicellular cyanobacterial strains.

Plant Terpene-Induced Expression of Multiple Aromatic Ring Hydroxylation Oxygenase Genes in Rhodococcus sp. Strain T104

  • Kim, Byung-Hyuk;Oh, Eun-Taex;Ahn, Yeong-Hee;Koh, Sung-Cheol
    • Journal of Microbiology
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    • v.41 no.4
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    • pp.349-352
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    • 2003
  • Recent studies have shown that some of the PCB (polychlorinated biphenyl)-degraders are able to effectively degrade PCB in the presence of monoterpenes, which act as inducers for the degradation pathway. Rhodococcus sp. T104, an effective PCB degrader, has been shown to induce the degradation pathway by utilizing limonenes, cymenes, carvones, and pinenes as sole carbon sources which can be found in the natural environment. Moreover, the strain T104 proved to possess three separate oxidation pathways of limonene, biphenyl, and phenol. Of these three, the limonene can also induce the biphenyl degradation pathway. In this work, we report the presence of three distinct genes for aromatic oxygenase, which are putatively involved in the degradation of aromatic substrates including biphenyl, limonene, and phenol, through PCR amplification and denaturing gradient gel electrophoresis (DGGE). The genes were differentially expressed and well induced by limonene, cymene, and plant extract A compared to biphenyl and/or glucose. This indicates that substrate specificity must be taken into account when biodegradation of the target compounds are facilitated by the plant natural substrates.