• Title/Summary/Keyword: ELISA antibody

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Identification and Cloning of a Fraction 1 Protein of Yersinia pestis that Produces Protective Immune Responses

  • Kim Jong-Hyun;Cho Seung-Hak;Jang Hyun-Chul;Lee Hee-Cheul;Kim Young-Il;Kang Yeon-Ho;Lee Bok-Kwon
    • Journal of Microbiology and Biotechnology
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    • v.16 no.8
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    • pp.1180-1184
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    • 2006
  • The capsule that surrounds Yersinia pestis cells is composed of a protein-polysacchride complex; the purified protein component is fraction I (F1) antigen. We report the cloning of the cafl gene and its expression in Escherichia coli using the vector pETl02/D-TOPO and the F1-specific monoclonal antibody. The recombinant F1 (rF1) antigen had a molecular size of 17.5 kDa, which was identical to that of the F1 antigen produced by Y. pestis. Recombinant F1 protein was found to react to polyclonal antiserum to Y. pestis Fl. Recombinant F1 was purified by ProBond purification system and induced a protective immune response in BALB/c mice challenged with up to 10$^5$ virulent Y. pestis. Purified rF1 protein was used in an ELISA to evaluate the ability of a method to detect antibodies to Y. pestis in animal sera. These results strongly indicated that the rF1 protein is a suitable species-specific immunodiagnostic antigen and vaccine candidate.

Characterization of a Cell Line HFH-T2, Producing Viral Particles, from Primary Human Fetal Hepatocytes Infected with Hepatitis B Virus

  • Shim, Jae-Kyoung;Kim, Dong-Wook;Chung, Tae-Ho;Kim, June-Ki;Suh, Jeong-Ill;Park, Chun;Lee, Young-Choon;Chung, Tae-Wha;Song, Eun-Young;Kim, Cheorl-Ho
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.186-192
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    • 2001
  • A primary culture of human fetal hepatocytes was obtained through a therapeutic abortion process at 26 weeks of gestation period. More than $10^8$ cells were seeded on a plastic plate. These hepatocytes were infected with hepatitis B virus (HBV). The HBV was purified from serum of one chronic HBV carrier. Transformed hepatocytes were subcultured in a 10% FBS-supplemented medium. The morphology of the transformed cell was epithelial-like. The cells from the first pass showed signs of early proliferation and had a latent period of more than 3 months after 6-7 passages. After the rest period, the transformed cell proliferated actively and they were subcultured every three days. Transformed hepatocytes were characterized by detection of the HBV transcript by RT-PCR. The secretion of virions from transformed cells was investigated by PCR with the cell medium. Two types of virions secreted into the culture medium were examined by using the transmission electron microscope. Another approach to study the secretion of virions in to culture medium was carried out with HBV antibody. HBsAg was detected in the culture medium of transformed cells using ELISA and Western blot analyses. These data suggested that the human fetal hepatocyte cell line has been established by infection of HBV, in which this cell line secreted viral particles into the culture medium.

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Characterization of the Physical Form of Allergenic Cry j 1 in the Urban Atmosphere and Determination of Cry j 1 Denaturation by Air Pollutants

  • Wang, Qingyue;Morita, Jun;Gong, Xiumin;Nakamura, Shinichi;Suzuki, Miho;Lu, Senlin;Sekiguchi, Kazuhiko;Nakajima, Takuya;Nakajima, Daisuke;Miwa, Makoto
    • Asian Journal of Atmospheric Environment
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    • v.6 no.1
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    • pp.33-40
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    • 2012
  • In this study, we characterized the physical form of allergenic Cry j 1 in the urban atmosphere. Through an immunofluorescence antibody method, we showed that allergenic Cry j 1 exists as fine particles (${\leq}1.1{\mu}m$). To determine Cry j 1 concentrations and its particle size distribution, we used the ELISA method to confirm that most Cry j 1 exists as fine particles in the urban atmosphere and is found at high concentrations on fine day next to rainy day. Furthermore, we evaluated Cry j 1 denaturation by using the Biacore J system based on the surface plasmon resonence (SPR) principle and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). We showed that the dissociation constant ($K_D$) of Cry j 1 that has been exposed to urban polluted air is lower ($1.76{\times}10^{-14}$ M) than that of Cry j 1 ($1.32{\times}10^{-9}-3.37{\times}10^{-9}$ M) of original pollen grains that has not been exposed to air pollutants. Cry j 1 turns into low molecular weight proteins by reacting with various acidic solutions. In sum, we showed that allergenic Cry j 1 exists as fine particles that can deposit in the lower respiratory tract. This finding clarifies the relationship between Japanese cedar pollinosis and air pollutants.

Prevalence of Human Papillomavirus and Herpes Simplex Virus Type 2 Infection in Korean Commercial Sex Workers

  • Yun, Hae-Sun;Park, Jeong-Joo;Choi, In-Kyung;Kee, Mee-Kyung;Choi, Byeong-Sun;Kim, Sung-Soon
    • Journal of Microbiology and Biotechnology
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    • v.18 no.2
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    • pp.350-354
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    • 2008
  • In order to investigate the prevalence of sexually transmitted viruses such as human papillomavirus (HPV) and herpes simplex virus (HSV) in Korean commercial sex workers (CSWs), we selected 188 CSWs (age range 20-44 years, median age 24 years) who regularly visited one public health center in Seoul, Korea. HPV genotypes were analyzed by using a HPV DNA Chip, and an enzyme-linked immunosorbent assay (ELISA) was used to detect type-specific IgG against HSV2 antibody identifying seropositivity for HSV2 infection. Polymerase chain reaction (PCR) was performed with specific primers to detect HPV and HSV1/2 in cervical swabs from the CSWs. The prevalence of HPV infection was 83.5% in 188 cervical swab specimens and the main high-risk HPV genotypes were HPV16, 18, 56, and 58. The principal low-risk HPV genotypes were HPV6 and 11. The prevalence of HSV1/2 DNA was 13.8% and HSV2 seroprevalence was 86.2%. These results suggest that high frequencies of HPV and HSV2 infection might contribute to the rapid spread of STD viruses in CSWs in Korea. Additionally, an understanding of why high-risk HPV genotypes are so prevalent could provide guidelines for prophylactic vaccine development in Korea.

Development of Quantitative Vitellogenin ELISAs for Bullfrog (Rana catesbeiana) used in Endocrine Disrupter Screening (내분비계 장애물질 검색을 위한 효소면역측정법을 이용한 황소개구리 비텔로제닌 정량법 개발)

  • Lee Sang-Hoon;Kang Yun-Ju;Li Chun-Ri;Kim Andre;Jin Chun-Feng;Chung Kyu-Hyuck;Kim Dong-Kyoo;Park Nam-Gyu;Park Kwang-Sik;Kang Shin-Won;Park Jang-Su
    • Environmental Analysis Health and Toxicology
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    • v.21 no.2 s.53
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    • pp.147-151
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    • 2006
  • 난생 생물의 알 생성유도 단백질인 비텔로제닌(viteILogenin, VTG)을 성숙한 암컷 황소개구리 (Rana catesbeiana)혈청으로부터 음이온 교환 크로마토그래피를 이용하여 정제 하였으며 정제한 비텔로제닌을 BALB/c mice에 주사하여 폴리크로날 항체를 생산하였고 이것은 protein A column으로 정제 하였다. 이렇게 정제된 폴리크로날 항체를 이용하여 황소개구리 비탈로제닌 측정용 효소면역측정법을 개발하였으며 그 측정 범위는 $12{\sim}1,560ng/mL$였다. 또한 이 효소면역측정법을 평가하기 위해 성숙한 수컷 황소개구리를 청정지역과 폐수처리장 하류 하천에서 서식하는 황소개구리 혈액 속의 비텔로제닌을 측정하였다. 그 결과 폐수처리장 하류 하천에 서식하는 수컷 황소개구리 비텔로제닌이 청정지역보다 현저하게 높게 유도됨을 알 수 있었다.

The Safety of Brucella abortus strain RB51 in pregnant cattle (부루세라 RB51 백신의 임신소에 대한 안전성)

  • Kim, Jong-man;Woo, Sung-ryong;Lee, Ji-youn;Jung, Suk-chan;Jean, Young-hwa;Kim, Jae-hoon;Kweon, Chang-hee;Yoon, Young-dhuk;Yoo, Han-sang
    • Korean Journal of Veterinary Research
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    • v.43 no.4
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    • pp.667-675
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    • 2003
  • The safety of Brucella abortus strain RB51(SRB51) was investigated in dairy cows and Korean native cattle of 4~7th month of gestation. From experimentally inoculated cattle, 18 of 25 (72.0%) dairy cows, and 3 of 10 (30.0%) Korean native cattle were aborted or delivered premature fetus. There were no significant differences in the incidence of abortion depending on the inoculation route (intramuscular and subcutaneous) and dosages ($1{\times}10^9$, $2.8{\times}10^9$, and $4.0{\times}10^9$ CFU). The antibodies to the SRB51 were measured by a dot blot enzyme-linked immunosorbent assay. The highest titers to SRB51 were detected between 5~7 weeks after inoculation and the specitic antibody could be detected up to 28 weeks after inoculation. The SRB51 was isolated from amnio-allantoic fluid, bronchial lymph node, mammary gland, and supramammary lymph node in 5 of 25 dairy cows during 4 weeks after either abortion or delivery. Although SRB51 was isolated from 4 of 24 aborted fetus or normally delivered calves at parturition time, it was not isolated during 4 weeks afterward. Eleven of twentyfive dairy cows showed the endometritis and/or necrosis until 6 weeks after delivery, no lesions were seen at 8 weeks after delivery and uterus from control dairy cows. The results of present study revealed that SRB51 might induce the clinical signs of brucellosis in the pregnant cattle at 4~7th month of gestation.

Acute porcine reproductive and respiratory syndrome outbreaks in immunized sow herds: from occurrence to stabilization under whole herd vaccination strategy

  • Moon, Sung Ho;Yoo, Sung J.;Noh, Sang Hyun;Kwon, Taeyong;Lee, Dong Uk;Je, Sang H.;Kim, Myung Hyee;Seo, Sang Won;Lyoo, Young S.
    • Korean Journal of Veterinary Research
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    • v.58 no.2
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    • pp.73-79
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    • 2018
  • Outbreaks of porcine reproductive and respiratory syndrome virus (PRRSV) in vaccinated sow herds from occurrence to stabilization were monitored and analyzed in terms of serology and reproductive performance. Three different conventional pig farms experienced severe reproductive failures with the introduction of a type 1 PRRSV. These farms had adopted mass vaccination of sows using a type 2 PRRSV modified live vaccine (MLV). Therefore, to control the type 1 PRRSV, an alternative vaccination program utilizing both type 1 and type 2 MLV was undertaken. Following whole herd vaccinations with both types of MLV, successful stabilization of PRRS outbreaks was identified based on serological data (no viremia and downward trends in ELISA antibody titers in both sows and suckling piglets) and recovery of reproductive performance. Additionally, through comparison of the reproductive parameters between outbreak and non-outbreak periods, it was identified that PRRSV significantly affected the farrowing rate and the number of suckling piglets per litter at all three pig farms. Comparison of reproductive parameters between periods when the different vaccination strategies were applied revealed that the number of piglets born in total and born dead per litter were significantly increased after the introduction of the type 1 PRRS MLV.

Seroreactivity to Helicobacter pylori Antigens as a Risk Indicator of Gastric Cancer

  • Karami, Najmeh;Talebkhan, Yeganeh;Saberi, Samaneh;Esmaeili, Maryam;Oghalaie, Akbar;Abdirad, Afshin;Mostafavi, Ehsan;Hosseini, Mahmoud Eshagh;Mohagheghi, Mohammad Ali;Mohammadi, Marjan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.3
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    • pp.1813-1817
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    • 2013
  • Background: Multiple etiologic factors are suspected to cause gastric cancer, the most important of which is infection with virulent types of Helicobacter pylori. Materials and Methods: We have compared 102 gastric cancer patients with 122 non-ulcer, non-cancer dyspeptic patients. Gastric specimens were evaluated for H. pylori infection by tissue-based detection methods. Patient sera underwent antigen-specific ELISA and western blotting using a Helicoblot 2.1 kit and antibody responses to various H. pylori antigens were assessed. Results: The absolute majority (97-100%) of both groups were H. pylori seropositive. Multivariate regression analysis demonstrated serum antibodies to the low molecular weight 35kDa protein to be protective and reduce the risk of gastric cancer by 60% (OR:0.4; 95%CI:0.1-0.9). Conversely, seroreactivity to the 89kDa (VacA) protein was significantly higher in gastric cancer patients (OR:2.7; 95%CI:1.0-7.1). There was a highly significant association (p<0.001) between seroreactivity to the 116kDa (CagA) and 89kDa (VacA) proteins, and double positive subjects were found at nearly five fold (OR:4.9; 95%CI:1.0-24.4) enhanced risk of gastric cancer as compared to double negative subjects. Conclusions: Seroreactivity to H. pylori low (35kDa) and high (116kDa/89kDa) molecular weight antigens were respectively revealed as protective and risk indicators for gastric cancer.

Insights into the Incidence of Watermelon chlorotic stunt virus Causing Yellowing Disease of Watermelon in Western and Southwestern Regions of Saudi Arabia

  • Ahmad, M.H.;Shakeel, M.T.;Al-Shahwan, I.M.;Al-Saleh, M.A.;Amer, M.A.
    • The Plant Pathology Journal
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    • v.34 no.5
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    • pp.426-434
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    • 2018
  • During the spring season of 2014, a total of 148 melon and watermelon leaf samples were collected from symptomatic and asymptomatic plants in the western and southwestern regions of Saudi Arabia and were tested for the presence of Watermelon chlorotic stunt virus (WmCSV) and other suspected cucurbit viruses by double antibody sandwich enzyme-linked immunosorbent assays. Ninety-eight samples were found to be positive for the presence of WmCSV, nine samples were positive for the presence of Cucurbit yellow stunting disorder virus (CYSDV), and 22 showed a mixed infection with both WmCSV and CYSDV. No other cucurbit viruses were detected in any of the samples. Host range experiments revealed that eight out of fourteen tested plant species were susceptible to WmCSV. PCR products of approximately 1.2 kb were obtained after amplification using primers specifically targeting the coat protein region of WmCSV. Positive PCR results were confirmed by dot blot hybridization. Coat protein gene sequences from eleven WmCSV isolates indicated that the highest identity was between the 104WMA-SA isolate from the Wadi Baish location and a previously reported isolate from the AL-Lith location in Saudi Arabia. The lowest identity was observed between the 42WMA-SA isolate and an isolate from Palestine.

Presumptive Diagnosis of Mycoplasma pneumoniae Pneumonia in Children (소아에서 마이코플라즈마 폐렴의 예기적 진단)

  • Lee, Chang Eon;Park, Su Jin;Kim, Won Duck
    • Journal of Yeungnam Medical Science
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    • v.29 no.2
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    • pp.89-95
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    • 2012
  • Background: As Mycoplasma pneumoniae pneumonia has increased in Korea, its relevance to infants, toddlers, and adolescents has magnified as well as. However, it is difficult to perform the serological test and PCR test routinely for diagnosis in actual clinical practice. Thus, the authors conducted this study to help clinicians do presumptive diagnosis of Mycoplasma pneumoniae pneumonia using clinical, radiological, and hematological findings. Methods: The study population consisted of 224 children between 1 month and 14 years old, hospitalized for radiographically confirmed pneumonia. Patients were divided into two groups of 100 children with Mycoplasma pneumoniae pneumonia, as diagnosed using the ELISA method. Groups with negative result in Mycoplasma IgM antibody test were classified into the viral group (98 patients with respiratory virus) and the bacterial group (46 patients with the bacteria detected in the blood sputum culture or antibiotic treatment except macrolide improved the patient's condition). These groups were compared and analyzed using clinical, hematological, and radiographic differences and scoring system. Results: Clinical, hematological, and radiographic characteristics of Mycoplasma pneumoniae pneumonia have shown the intermediate level results between bacterial pneumonia and viral pneumonia. In terms of scoring system, the mean score of Mycoplasma pneumoniae pneumonia was 4.23, which was the intermediate level between bacterial pneumonia (mean score=6.67) and viral pneumonia (mean score=1.48). Conclusion: Results suggest that the combination of the scoring system information can increase the accuracy in the diagnosis even if they may have difficulties on diagnosis, because clinical manifestations, hematological, and radiographic findings are nonspecific.

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