• Title/Summary/Keyword: antigen test

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Expression and diagnostic application of p12 protein of African swine fever virus by recombinant baculovirus (재조합 baculovirus에 의한 아프리카 돼지콜레라바이러스 p12 단백질의 발현과 진단적 적용)

  • Choi, Kang-Seuk;Choi, Cheong-up;Kim, Yong-Joo
    • Korean Journal of Veterinary Research
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    • v.45 no.1
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    • pp.63-70
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    • 2005
  • African swine fever (ASF) is an infectious disease of domestic and wild pigs for which there is no vaccine in the world. A proper surveillance of viral activity and a timely response to ASF outbreaks depend upon the rapid diagnosis of ASF viral infection. Internationally prescribed enzyme-linked immunosorbent assay (ELISA) is a fast, sensitive test routinely used in the diagnosis of the ASF. However, inactivated whole ASF virus antigen used in this test is a tedious to prepare and has a risk of outside exposure of infectious virus by laboratory accident during the preparation. An ASF virus noninfectious recombinant antigen is a safe and easily produced alternative antigen for use in diagnostic assay. We have cloned the ORF O61R gene of the ASF virus to generate a recombinant baculovirus producing the p12 protein in insect cells under control of the polyhedrin promoter as non-fusion protein. When used in an indirect ELISA, the p12 antigen showed reactivity with all known ASF positive pig sera but not with negative pig sera. Our results indicated that the p12 protein would be one of alternative antigens for diagnosis of the ASF.

Clearance of False-positive Antigen-Antibody Reactions of a Diagnostic Antigen Production in Escherichia coli with Human Sera

  • Noh, Kap-Soo;Kim, Jong-Wan;Ha, Suk-Hoon;Yoo, Wang-Don;Jeon, Weong-Joong;Kim, Hyun-Su
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.4 no.1
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    • pp.63-65
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    • 1999
  • Although many pharmaceutically useful proteins are produced in E. coli expression system, it is very are rare for the system to be used in the production of diagnostic antigen due to a major problem, i.e., false-positive reaction of e. coli host-derived proteins contaminating purified diagnostic antigen with human sera. The N (nucleocapsid) protein of Seoul virus causing haemorrhagic fever with renal syndrome (HFRS) was produced in E. coli BL21 (DE3), and used for the detection of N protein-specific antibodies in human sera. Using the N protein as a diagnostic antigen of HFRS, the false-positive reaction was cleared by merely mixing the test sera with the extract of E. coli host strain not harboring expression plasmid.

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Expression of Hepatitis B Virus Antigen by Recombinant Vaccinia Virus VV-$\textrm{HBV}_{L}$

  • Lee, Yun-Kyung;Yu, Jung-An;Ahn, Byung-Yoon;Aree Moon
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1997.04a
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    • pp.82-82
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    • 1997
  • The hepatitis B virus(HBV) is a small, enveloped virus with a circular, double-stranded DNA genome. HBV causes active and chronic hepatitis worldwide, including Korea, and is considered to be a major factor for liver cirrhosis and hepatocellular carcinoma. In contrast to the wealth of knowledge on the gene structure and expressional regulation, immunological and pathological mechanisms for HBV-induced hepatocellular injury are not well known. In the present study, vaccinia virus which has been demonstrated to be a useful eukaryotic expression vector was used to clone the gene for HBV surface antigen, L(S+preS2+preS1). The recombinant vaccinia virus vector, pMJ-L, which contains L surface antigen gene of adr-type HBV was constructed, and subseouently used for making recombinant vaccinia virus VV-$\textrm{HBV}_{L}$. Expression of the HBV antigen was examined by immunofluorescent antibody (IFA) test using mouse monoclonal anti-hepatitis B surface antigen. HBsAg was detected in the recombinant virus indicating that the VV-$\textrm{HBV}_{L}$ expressed S antigen successfully. The HBV-Vaccinia Virus recombinant obtained in this study is currently being used for studying the immunological aspects of HBV infection.

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An Improved Method for Detection of Salmonella Typhi O Antigen with Staphylococcal Protein A Using Enzyme Immunoassay (포도구균의 A단백질을 이용한 효소면역법으로 살모넬라 O항원 검출)

  • Rhyu, Mun-Gan;Kim, Gum-Ryong;Lee, Choong-Ki
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.4
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    • pp.445-451
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    • 1987
  • Coagglutination method is widely used for the diagnosis of Salmonella infection. This test, however, has a disadvantage of false positive reaction due to the coagglutination of staphylococci with non-specific immune complexes or anti-staphylococci antibody in serum. Salmonell O antigen was detected by enzyme immunoassay with protein A-bearing Staphylococcus aureus as in the solid phase. Horse radish peroxidase was labeled to IgG specific against Salmonella O antigen. This enzyme immunoassay was much more sensitive than conventional coagglutination method without false poitive agglutination. To improve the sensitivity for detection of Salmonella O antigen in samples, we tried to determine the optimal concentration of normal IgG that inhibits non-specific binding of horse radish peroxidase labeled IgG to staphylococci, and to establish the optimal condition of reaction between antigen-antibody complex and staphylococci. Non-specific binding of horse radish peroxidase labeled specific IgG to staphylococci was almost blocked when the enzyme labeled IgG was 500-fold diluted with phosphate buffered saline containing 2mg/ml of normal IgG. When staphylococci coated with antibody to Salmonella O antigen were mixed with antigen-antibody complex and then incubated for 1 hour at room temperature, the minimal detectable concentration of Salmonella O antigen was 1ng/ml. The sensitivity of enzyme immunoassay was 100-fold greater than a conventional coagglutination method. This enzyme immunoassay could be expected as an improved method for detection of other infectious agents.

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Clinical usefulness of rapid antigen test to detect respiratory syncytial virus infection (Respiratory syncytial virus 감염진단을 위한 신속항원검사의 유용성)

  • Kim, Hyung Su;Kim, Hee La;Park, Ki Hyung;Cho, Kyung Soon
    • Clinical and Experimental Pediatrics
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    • v.51 no.10
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    • pp.1071-1076
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    • 2008
  • Purpose : Respiratory syncytial virus (RSV) is the most frequent cause of lower respiratory infections in infants and young children. Early detection allows quarantining of infected inpatients to prevent nosocomial transmission and to choose a treatment. To achieve rapid reporting, to facilitate prompt antiviral therapy, and to avoid unnecessary use of antibiotics, an easy, rapid diagnostic method for RSV is needed. We evaluated a lateral flow immunochromatography (RSV Respi-Strip test) and EIA (Enzyme immuno assay) compared to RT-PCR. Methods : From April 2007 to March 2008, 112 consecutive respiratory specimens (nasopharyngeal aspirates, throat swabs, tracheal aspirates, sputum) from patients who were suffering from the clinical signs and symptoms of respiratory tract infection were enrolled in Busan. A total of 112 patients were tested with RSV Respi-Strip (Corio-BioConcept, Belgium), EIA, and RT-PCR at the same time. Results : Of the 112 specimens tested, the number of children who showed positive results at RT-PCR and Respi-Strip were 45 and 42, respectively. The Respi-Strip rapid antigen test had a sensitivity of 88% and a specificity of 94%. The positive and negative predictive values were 90% and 92%, respectively. The agreement was 83%. Conclusion : In our study, the rapid antigen test had as much sensitivity as any method for detection of RSV. The test has many advantages such as easy performance, simple interpretation, and rapid results. If the rapid antigen test is widely applied in the clinical setting, the may be useful for diagnostic and epidemiological studies of RSV infection.

Comparison of the non-invasive diagnostic methods, stool antigen test and PCR assay, for Helicobacter felis detection in dogs

  • Hong, Sunhwa;Lee, Hak-Yong;Kim, Tae-Wan;Kim, Okjin
    • Korean Journal of Veterinary Service
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    • v.38 no.1
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    • pp.37-42
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    • 2015
  • The aim of the present study was to compare the non-invasive methods for the diagnosis of H. felis with HpSA kit-based detection method and H. felis-specific PCR assay with dog's stool samples without sacrifice. Male Beagle dogs (n=6) were infected with H. felis ATCC 49179 ($1.0{\times}10^9CFU/dog$) by intra-gastric inoculation two times at 3-day intervals, and the stool specimens of dogs were collected 1, 3, 5, 7, 14, 21 days after infection to submit to HpSA test and H. felis-specific PCR. As the results, the sensitivity of the HpSA and the PCR analysis was 50.0%, 83.3% respectively. Although HpSA test is less sensitive, it could be used for rapid, cheap and easy screening assay for H. felis infection in dog and cats. We suggest that the H. pylori stool antigen kit, HpSA, is useful and effective for monitoring H. felis infection. If HpSA test would be made with H. felis antibodies in the future, its sensitivity could be increased. Also, PCR assay could be successfully used to detect the H. felis in stools. Applying the H. pylori stool antigen kit and PCR assay may be the recommended non-invasive strategy to identify H. felis in dog and cats.

A carbohydrate antigen of Clonorchis sinensis recognized by a species-specific monolonal antibody (종특이 단세포군항체에 반응하는 간흡충의 당질항원 한 가지)

  • 용태순;이종석
    • Parasites, Hosts and Diseases
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    • v.34 no.4
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    • pp.279-282
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    • 1996
  • The enzyme-linked immunosorbent assay (ELISA)-inhibition test using a Clonorchis sinensis species-specific mouse monoclonal antibody (MAb) , CsHyb 0605-23, showed increased specificity over the conventional ELISA used for serodiagnosis of clonorchiasis. To characterize the corresponding antigen further, the MAb was tested against polysaccharide, protein and glycolipid fractions obtained from a crude extract of C. slnensis adult worms, using chloroform, methanol and phenol extractions. Only the polysaccharide fraction was recognized by the mb among those fractions. Mild oxidation of the antigen with sodium periodate showed decreased reactivity against the MAb. We concluded that the antigen and antigenic determinants recognized by the MAb are carbohydrates.

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Usefulness of the Milk Ring Test for the Diagnosis of Brucellosis in Dairy Cattle (유우(乳牛)브루셀라병(病) 진단(診斷)을 위(爲)한 Milk Ring Test의 이용가치(利用價値))

  • Chung, Byung Tack
    • Korean Journal of Veterinary Research
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    • v.9 no.2
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    • pp.55-59
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    • 1969
  • For the establishment of simpler and more reliable methods for the diagnosis of Brucellosis, diagnostic antigen for Milk Ring Test (M.R.T.) was prepared according to working document recommended by Ministry of Agriculture U.S.A., and applied in the field for the test of its's applicability. The results are summarized as follows. 1. Of fifty two dairy cattle ranches in the vicinity of Seoul, five ranges were shown positive reaction to M.R.T. 2. Even though one volume of positive milk was mixed with five to ten volumes of negative milk, the positive reactions were also observed. 3. Antigenicity of the antigen was maintained without deteriolation for 13 months at $5^{\circ}C$ and three months at both $22^{\circ}C$ and $37^{\circ}C$. 4. Three positive cows to M.R.T. were also shown suspective reaction by Serum Agglutination Test.

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Fusion Analytical Sensitivity of Rapid Influenza Antigen Limit of Detection Tests for Human Influenza virus (인플루엔자 바이러스에 대한 신속 항원 검출 검사 검출한계의 융합적 분석)

  • Song, Chang-Sub;Sung, Hyun-Ho;Kim, Jung-Hyun;Kim, Dae-Eun;Park, Chang-Eun;Yoon, Joong-Soo
    • Journal of the Korea Convergence Society
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    • v.9 no.3
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    • pp.165-171
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    • 2018
  • In this study, to analyze the detection of limit for sensitivity of the influenza rapid antigen test kit, the positive detection of limits were analyzed by serial dilution of influenza virus A and B type for five influenza rapid antigen test kits in Korea. As a result of analysis, visual measurement of type A were up to 1:8192 for the Wellsbio product and up to 1:4096 for the II product, up to 1:512 for the I and III products, and only 1:128 for the IV product, and type B were positive for up to 1:8192 for the Wellsbio product, up to 1: 4096 for the II product and up to 1:1024 for the I, III and IV products. For instrument readings with the same specimen, both A and B types were found to be positive for up to 1:8192 for the Wellsbio product, up to 1: 4096 for the II product, and up to 1:2048 for the I product. The sensitivity of the rapid antigen test for influenza differs greatly depending on the sampling area of the patient, infection period, specimen volume, etc. Therefore, it is necessary to observe exactly the collection timing and method of the specimen. And it is necessary further study to improve the sensitivity for influenza rapid antigen test.

Study on prevalence of antigens to bovine viral diarrhea virus (BVDV) of Cattle in Busan area (2013~2014) (부산지역 소 바이러스성 설사병 바이러스(bovine viral diarrhea virus; BVDV) 감염 실태 조사·연구(2013~2014))

  • Kim, Hong-Tae;Park, Min-Sik;Lee, Gi-Heun;Lee, Keun-Woo
    • Korean Journal of Veterinary Service
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    • v.38 no.1
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    • pp.43-49
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    • 2015
  • Bovine viral diarrhea virus (BVDV) is a very important viral disease virus in cattle, domestic and wild ruminants. The purpose of this study is to investigate the positive rate of bovine viral diarrhea virus antigen by ELISA from Korean native and beef cattle reared in Busan area from March in 2013 to October in 2014. A total of 1,129 bovine blood samples were collected from 140 farms, 1,111 Korean native cattle of 135 farms and 18 beef cattle of 5 farms. Test for antigen was carried out by ELISA method. In general analysis, the positive rate of bovine viral diarrhea virus antigen were 0.7% (8/1,129) cattle and 5.0% (7/140) farm. In regional analysis, the positive rate of BVDV antigen of farm in Kijang-gun, Gangseo-gu, Geumjeong-gu, Saha-gu and Dongnae-gu were 1.4% (2/94), 3.6% (5/37), 0% (0/7), 0% (0/1) and 0% (0/1), respectively, and the positive rate of BVDV antigen of cattle were 0.4% (3/770), 1.5% (5/333), 0% (0/24), 0% (0/1) and 0% (0/1), respectively. The positive rate of BVDV antigen according to sex were 0.6% (6/1,085) female cattle and 4.6% (2/44) male cattle. According to the age of cattle, the positive rate of BVDV antigen in 1 year, 2 years, 3 years and 5 years old were 1.9% (4/215), 0.4% (1/265), 0.9% (2/234) and 1.0% (1/103), respectively, but 4 years (0/198), 6 years (0/55), 7 years (0/24), 8 years (0/14), 9 years (0/10), 10 years (0/7) and 11-15 years (0/3) old were negative, respectively.