• 제목/요약/키워드: plaque assay

검색결과 141건 처리시간 0.029초

Improvement of indirect enzyme-linked immunosorbent assay for detection of Japanese encephalitis virus antibodies in swine sera

  • Yang, Dong-Kun;Kim, Ha-Hyun;Jo, Hyun-Ye;Lee, Seung Heon;Jang, Sang-Ho;Lee, Sang-Oh;Choi, Sung-Suk;Cho, In-Soo
    • 대한수의학회지
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    • 제57권1호
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    • pp.31-36
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    • 2017
  • Japanese encephalitis (JE) is an important zoonosis caused by the mosquito-transmitted JE virus (JEV), which is a causative agent of reproductive failure in pregnant sows. Detection of JEV antibodies in swine is performed by hemagglutination inhibition (HI), virus neutralization (VN), and the plaque reduction neutralization test (PRNT). The most stringent PRNT is the 90% endpoint PRNT ($PRNT_{90}$). These conventional assays are difficult to carry out in diagnostic laboratories with insufficient instruments or cell culture systems. An alternative assay that is easily conducted and time efficient is required. In this study, we improved the indirect enzyme-linked immunosorbent assay (I-ELISA) with clarified antigen for the detection of JEV antibodies. The I-ELISA results obtained from 175 swine serum samples were compared with HI, VN, and $PRNT_{90}$ results. The sensitivity of I-ELISA was 91.8%, 95.0%, and 94.7% compared with HI, VN, and $PRNT_{90}$ results, respectively. The specificity of I-ELISA was 92.2%, 94.7%, and 94.7% compared with HI, VN, and $PRNT_{90}$ results, respectively. Moreover, the I-ELISA results were significantly correlated with the HI (r = 0.93), VN (r = 0.95), and $PRNT_{90}$ (r = 0.92) results. These results suggest that the improved I-ELISA is useful for serosurveillance of JEV in swine.

SARS-CoV-2 Antibody Neutralization Assay Platforms Based on Epitopes Sources: Live Virus, Pseudovirus, and Recombinant S Glycoprotein RBD

  • Endah Puji Septisetyani;Pekik Wiji Prasetyaningrum;Khairul Anam;Adi Santoso
    • IMMUNE NETWORK
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    • 제21권6호
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    • pp.39.1-39.18
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    • 2021
  • The high virulent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus that emerged in China at the end of 2019 has generated novel coronavirus disease, coronavirus disease 2019 (COVID-19), causing a pandemic worldwide. Every country has made great efforts to struggle against SARS-CoV-2 infection, including massive vaccination, immunological patients' surveillance, and the utilization of convalescence plasma for COVID-19 therapy. These efforts are associated with the attempts to increase the titers of SARS-CoV-2 neutralizing Abs (nAbs) generated either after infection or vaccination that represent the body's immune status. As there is no standard therapy for COVID-19 yet, virus eradication will mainly depend on these nAbs contents in the body. Therefore, serological nAbs neutralization assays become a requirement for researchers and clinicians to measure nAbs titers. Different platforms have been developed to evaluate nAbs titers utilizing various epitopes sources, including neutralization assays based on the live virus, pseudovirus, and neutralization assays utilizing recombinant SARS-CoV-2 S glycoprotein receptor binding site, receptor-binding domain. As a standard neutralization assay, the plaque reduction neutralization test (PRNT) requires isolation and propagation of live pathogenic SARS-CoV-2 virus conducted in a BSL-3 containment. Hence, other surrogate neutralization assays relevant to the PRNT play important alternatives that offer better safety besides facilitating high throughput analyses. This review discusses the current neutralization assay platforms used to evaluate nAbs, their techniques, advantages, and limitations.

전기분해수소수의 Streptococcus mutans에 대한 항균효과 (Antibacterial effect of electrolyzed water on Streptococcus mutans)

  • Kim, Ji-Hye;Youn, Ha-Young;Kim, Eun-Kyong;Lee, Young-Eun;Jang, Ji-Eon;Song, Keun-Bae
    • 한국치위생학회지
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    • 제21권5호
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    • pp.527-533
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    • 2021
  • 연구목적: 본 연구는 대표적인 치아우식 유발 세균인 Streptococcus mutans(S. mutans)에 대한 전기분해 수소수의 항균효과와 치면세균막 억제 효과를 평가하기 위해 실시하였다. 연구방법: BHI 액체배지 상에서 6시간 배양한 S. mutans를 전기분해수소수에 1분 또는 3분 노출시켰다. 전기분해수소수의 증식억제 효과를 확인하기 위해 균주 현탁액을 Mitis Salivarius agar bacitracin(MSB) 배지 상에서 48시간 배양한 후 집락수를 계수하였으며, biofilm formation assay에 따라 crystal violet solution을 30분간 처리하여 치면세균막을 염색하고, 600 nm의 파장에서 흡광도를 측정하여 치면세균막 형성정도를 평가하였다. 연구결과: 전기분해수소수는 S. mutans의 증식과 치면세균막 형성을 유의하게 억제하였으며(p<0.001), 특히 치면세균막 형성의 경우 노출 시간이 증가함에 따라 치면세균막 형성도 유의하게 감소하였다(p<0.05). 결론: 수돗물 전기분해수소수는 S. mutans의 증식과 치면세균막 형성 억제를 통해 치아우식을 효과적으로 예방할 수 있다고 판단된다.

Validation of a Real-Time RT-PCR Method to Quantify Newcastle Disease Virus (NDV) Titer and Comparison with Other Quantifiable Methods

  • Jang, Juno;Hong, Sung-Hwan;Kim, Ik-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제21권1호
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    • pp.100-108
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    • 2011
  • A method for the rapid detection and quantification of Newcastle disease virus (NDV) produced in an animal cell culture-based production system was developed to enhance the speed of the NDV vaccine manufacturing process. A SYBR Green I-based real-time RT-PCR was designed with a conventional, inexpensive RT-PCR kit targeting the F gene of the NDV LaSota strain. The method developed in this study was validated for specificity, accuracy, precision, linearity, limit of detection (LOD), limit of quantification (LOQ), and robustness. The validation results satisfied the predetermined acceptance criteria. The validated method was used to quantify virus samples produced in an animal cell culture-based production system. The method was able to quantify the NDV samples from mid- or late-production phases, but not effective on samples from the early-production phase. For comparison with other quantifiable methods, immunoblotting, plaque assay, and tissue culture infectious dose 50 ($TCID_{50}$) assay were also performed with the NDV samples. The results demonstrated that the real-time RT-PCR method is suitable for the rapid quantification of virus particles produced in an animal cell-culture-based production system irrespective of viral infectivity.

솔잎, 돌나물, 톳, 메밀, 깻잎 등 5가지 혼합 열수 추출물의 면역 활성 효과 (Studies on the Effects of Water Extract from Mixture of Pine Needles, Sedum sarmentosum Bunge, Hijkiaorme, Buckwheat and Perlla Leaves on the Immune Function Activation)

  • 류혜숙;김현숙
    • 한국식품영양학회지
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    • 제21권3호
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    • pp.269-274
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    • 2008
  • Plants have long been used as a food source in Korea. In this study, we investigated the combined immunomodulative effects of a water extract mixture of(pine needles, Sedum sarmentosum Bunge, hijkiaorme, buckwheat and Peril a leaves) on Balb/c mice $7{\sim}8$ weeks old. The mice were fed a chow diet ad libitum and the plant extract was orally administered every other day for four weeks at two different concentrations(50 and 500 mg/kg BW). After preparing the single-cell suspension, splenocyte proliferation was determined by the MTT(3-[4,5-di-methylthiazol-2-y]-2,5-diphenyl terazolium bromide) assay. After 48hrs of incubation with the mitogens(ConA or LPS) splenocyte from the mice groups administered 50 and 500 mg/kg BW of the plant extract showed a significant increased in proliferation compared to the control group. A hemolytic plague forming cell assay was used to indicate antibody production against sheep red blood cells(SRBC). The number of antibody-secreting cells T-dependent antigen. The result of this study suggest that supplementation with this plant extract may regulate immune function by increasing splenocyte proliferation and the number of plaque forming cells.

Antiviral Triterpenes from Prunella vulgaris

  • Ryu, Shi-Yong;Lee, Chong-Kyo;Lee, Chong-Ock;Kim, Hae-Soo;Zee, Ok-Pyo
    • Archives of Pharmacal Research
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    • 제15권3호
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    • pp.242-245
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    • 1992
  • Two triterpenes 1 and 2 with antiviral activity against Herpes simplex virus type 1 in vitro were isolated from Prunella vulgaris. Each compound caused a significant reduction in viral cytopathic effect when vero cells were exposed to them for 72 hours after viral challenge. They were identified as betulinic acid (1) and $2\alpha, 3\alpha$-dihydroxyurs-12-en-28-oic acid(2) on the basis of their spectroscopic properties. The antiviral activity of them was estimated as $EC_{50}=30\;\mu$g/ml(1) and $8\;\mu$g/ml(2), respectively by plaque reduction assay.

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Immunomodulatory Response Induced by Ginseng

  • Kumar, Ashok
    • Journal of Ginseng Research
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    • 제27권3호
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    • pp.115-119
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    • 2003
  • There has been continuing interest in the development of synthetic and natural compounds that modify the immune response particularly for the treatment of AIDS and cancer. During the past fifty years, numerous scientific studies have been published on ginseng. Modem human studies have investigated preventive effect of ginseng on several kinds of cancer, its long term immunological effect on HIV patients, its effect on cell mediated immune functions in healthy volunteers. Similarly non clinical studies on animal model system have studied the chemopreventive action of ginseng on cancer and immunological properties of ginseng. The precise mechanism of action of ginseng, however, not clearly understood. Considering its wide-ranging therapeutic effects, this study is being undertaken to elucidate the general mode of action of ginseng, especially to test our hypothesis that its biological action may be mediated by the immune system.

Prevention of Murine Acquired Immunodeficiency Syndrome (MAIDS) Development by Oriental Herb Extracts

  • Yang, Yun-Hee;Yang, Joo-Sung
    • Journal of Applied Biological Chemistry
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    • 제48권4호
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    • pp.170-177
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    • 2005
  • Oriental medicinal herb extracts (OHE) showing anticancer activities were investigated for effectiveness as antiviral drugs. Infection of MuLV to cell line resulted in formation of giant syncytia. Number of giant syncytia in culture treated with OHE decreased by 40% compared to that of non-OHE-treated cell culture. To determine OHE effects on progeny release, RT-PCR was performed. In vivo animal studies demonstrated effectiveness of OHE as antiviral drug when administered orally. After OHE administration, viral cytopathic effects decreased. Infected mice showed splenomegaly and over-proliferation of lymphocytes with decreased CD4+ cell counts. These symptoms decreased in OHE-treated mice, indicating OHE maybe useful therapeutics against MuLV/MAIDS as Human Immunodeficiency Virus (HIV)/AIDS animal model. Results show XC plaque assay and in vivo MAIDS model using MuLV are suitable tools for screening anti-retroviral drug candidates.

Identification and Characterization of Bdellovibrio bacteriovorus, a Predator of Burkholderia glumae

  • Song, Wan-Yeob
    • Journal of Microbiology and Biotechnology
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    • 제14권1호
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    • pp.48-55
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    • 2004
  • Six strains of an obligate predatory bdellovibrio isolate that preys on Burkholderia glumae in rice paddy field water and rhizosphere soil, were identified and characterized. The numbers of Bdellovibrio cells varied from $3.2{\times}10^3$ to $9.2{\times}10^3$ plaque-forming unit/g after enrichment in cells of B. glumae. Prey range tests with six Bdellovibrio strains and 17 prey strains of rice-pathogenic, antibiosis-related, or nitrogen-fixing bacteria resulted in unique predation patterns in related prey cells. Strain BG282 had the widest prey range on 7 plant pathogenic bacteria among the 17 prey strains tested. However, no predation occurred with strains of Azospirillum brasilense, Paenibacillus polymyxa, Pseudomonas fluorescens, P. putida, and Serratia marcescens that are associated with antibiosis or nitrogen fixation in the rice ecosystem. Identification was confirmed by the presence of typical bdelloplast in the prey cells of B. glumae and by a PCR assay using B. bacteriovorus-specific primers. Furthermore, 16S rDNA sequencing of the six bdellovibrio strains showed a homology range of 97.2% to 99.2% to the type strain of B. bacteriovorus.

Effects of antitumor polysaccharides from albizza julibrissin on immune function

  • Moon, Chang-Kiu;Lee, Byeong-Gon;Lee, Soo-Hwan;Kang, Tak-Lim
    • Archives of Pharmacal Research
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    • 제8권4호
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    • pp.277-282
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    • 1985
  • Polysaccharide fractions were prepared from Albizzia julibrissin by different extraction schedules. The fractions obtained were designated as PS-I and PS-II, respectively. Further purification of PS-I by Sephadex G-200 column chromatography gave two subfractions, Alju A and alju B, Each fraction showed marked antitumor activity against sarcoma 180 solid form but not ascite form. PS-I and PS-II increased delayed hypersensitivity in normal and tumor bearing mice. PS-I treatment led to moderate restoration of the suppressed antibody production in tumor bearing mice.

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