• Title/Summary/Keyword: ELISA IgG

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Development of Sandwich ELISA for the Detection of Pork in Processed Foods (가공식품 중 돈육 검출을 위한 샌드위치 ELISA 개발)

  • Back, Su-Yeon;Do, Jeong-Ryong;Shon, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.47 no.3
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    • pp.401-404
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    • 2015
  • A sandwich ELISA (sELISA) to detect pork in processed foods was developed using goat anti-pig IgG antibodies. From the sELISA standard curve, the detection range of pork was $3-1,000{\mu}g/mL$. The cross-reactivity between the pig IgG antibodies, pork, and other meats (beef, chicken, fish, and crustaceas) was 100, 0.18, and 0%, respectively. When pork was heated for 10 min, the mean assay recoveries of pig-IgG were 79-32% at $60-70^{\circ}C$ and less than 0.11% at $80^{\circ}C$ or higher. When pork was spiked into cream soup, weaning food, fish paste, and sauce, the mean assay recoveries were 8.8, 45, 36, and 39%, respectively. In 12 commercial processed foods, the assay results coincided qualitatively with the food labels on the packages.

Application of Oral Fluid Sample to Monitor Porcine circovirus-2 Infection in Pig Farms (구강액을 이용한 양돈장의 Porcine circovirus-2 감염에 대한 모니터링)

  • Kim, Won-Il
    • Journal of Veterinary Clinics
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    • v.27 no.6
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    • pp.704-712
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    • 2010
  • Porcine circovirus-2 (PCV2) has been implicated in many clinical diseases/syndromes that are now referred to as PCV-associated diseases (PCVAD). Due to significant economic losses caused by PCVAD, many swine operations have launched extensive monitoring programs for PCV2. Traditional serum sampling is, however, rather expensive and laborious, hampering effective large scale pathogen surveillance. A field-based longitudinal study was conducted to assess the utility of pen-based oral fluid sample as an alternative to serum for herd PCV2 testing. Six pens (25 pigs/pen) at each of 3 different sites were used in the study. One oral fluid and 5 random serum samples per pen were collected at 3, 5, 8, 12, and 16 weeks of age, and the sera were pooled by pen for testing. All samples were tested for PCV2 by real-time PCR and for antibodies by indirect fluorescent antibody test (for both anti-PCV2 IgG and IgA) and 3 ELISA assays (blocking ELISA, indirect ELISA, and IgG/IgM sandwich ELISA). PCV2 DNA was detected in oral fluid samples sporadically until 8 weeks and in all pens at 16 weeks. PCV2-specific IgG was detected in oral fluid samples at 3 weeks and persisted until 5 to 8 weeks in all sites. Anti-PCV2 IgG and IgA were detectable in oral fluid samples collected at 16 weeks from all of the pens at 1 site. The detection of PCV2 and anti-PCV2 antibody in oral fluid samples correlated positively with results on pooled sera, suggesting that oral fluids can be a cost-effective alternative to serum for herd monitoring of PCV2 infection.

The Sensitivity Comparison of Immunodiagnostic Assays for Diagnosing Dengue Fever

  • Zahoor, Muhammad;Bahadar, Haji;Uddin, Salah;Naz, Sumaira
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.4
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    • pp.275-279
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    • 2016
  • Dengue fever is a vector borne disease caused by a dengue virus. It is an RNA virus of the family flaviviridae, with different serotypes. Herein, we report our attempt to carry out a sensitivity comparison of immunodiagnostic assays for dengue fever in dengue positive patients. Blood samples from 189 volunteers were collected. To determine the sensitivity of the NS1 test, two different types of tests-immunochromatographic tri-line test and rapid dengue test (RDT)-as well as IgM and IgG capture ELISA were performed. The result of RDT has shown that 59.7% of volunteers were IgM positive and 50.2% were IgG positive. Conversely, the results from capture ELISA shows 79.8% and 59.7% for IgM and IgG, respectively. The sensitivity of the capture ELISA test for IgM and IgG was higher than that of immunochromatographic tri-line rapid test, but the specificity was lower. Therefore, to confirm dengue fever, we recommend performing more detailed, investigative tests since a single test may not be sufficient.

Changes of IgM and IgG antibody levels in experimental rabbit anisakiasis as observed by ELISA and SDS-PAGE/immunoblot (ELISA와 SDS-PAGE/immunoblot을 이용한 실험적 토끼 아니사키스증에서 혈청 항체가의 변화)

  • Yang, Hyeon-Jong;Jo, Yu-Jeong;Baek, Yeong-Han
    • Parasites, Hosts and Diseases
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    • v.29 no.4
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    • pp.389-396
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    • 1991
  • Antibody changes in experimental anisakiasis were observed in 10 rabbits which were infected each with 10 Anisakis simplex larvae. The sera were collected before and on the 6th to the 95th day after the infection. Using crude saline extract of Anisakis larvae as antigen, specific IgM and IgG antibody levels were observed by ELISA and SDS-polyacrylamide gel electrophoresislimmunoblot. Levels of specific-IgM antibody were elevated from the 6th day, reached their peaks on the lIth day after the infection, and dropped thereafter. Serum levels of IgG antibody increased from the 6th day and reached their peak on the 26th day after the infection, and decreased gradually thereafter. When SDS-PAGE of the crude extract was done, at least forty-one SDS-polypeptide bands were recognized. Of them, IgM antibody reacted mainly to the bands of 168, 95, 74, 64, 51, 47 and 34 kDa while IgG antibody reacted strongly to 168, 92, 85, 64, 58, 52, 42 and 40 kDa bands. The crude extract showed negligible cross reactions with sera of other parasitic diseases and normal control. Key words: Anisakis simplex larvae, experimental anisakiasis, rabbit, antibody, ELISA, SDS- PAGE/immunoblot.

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Development of an Indirect Non-Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Cronobacter muytjensii in Infant Formula Powder (유아용 조제분유 내 Cronobacter muytjensii 검지를 위한 간접 비경합 면역분석법의 개발)

  • Song, Xinjie;Kim, Myunghee
    • The Korean Journal of Food And Nutrition
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    • v.26 no.4
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    • pp.936-944
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    • 2013
  • Cronobacter muytjensii is an important foodborne pathogen as a potential risk in infant formula powder (IFP). To develop a new and sensitive method for the detection of Cronobacter spp. in IFP, an immunoglobulin G (IgG) specific for C. muytjensii (formerly known as Enterobacter sakazakii ATCC 51329) was developed. Further, an indirect noncompetitive enzyme-linked immunosorbent assay (INC-ELISA) was developed by using the anti-C. muytjensii IgG. As a result, this newly developed INC-ELISA method was found very sensitive for C. muytjensii with detection limit of $6.5{\times}10^3CFU/ml$ in pure culture and 1 cell/25 g of IFP. This INC-ELISA method also displayed excellent specificity for C. muytjensii showing no cross-reactivity with other strains of Cronobacter genus and 11 other foodborne pathogenic strains. These results show that the developed INC-ELISA method was very sensitive, efficient, and rapid for the detection of C. muytjensii. Hence, this method could be applied to the development of diagnostic kits for the rapid and easy detection of C. muytjensii.

Development of Chemiluminescence Immunoassay(CIA) & ELISA for the Detection of Anti-sperm Antibodies in Male Serum (항정자 항체 검출을 위한 CIA 및 ELISA 개발을 위한 기초 연구)

  • Kim, S.C.;Lee, K.S.;Kim, Y.K.;Kim, C.K.;Choi, K.H.;Kwon, O.J.;Kim, J.B.
    • Clinical and Experimental Reproductive Medicine
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    • v.17 no.1
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    • pp.71-80
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    • 1990
  • New immunoassay systems for the detection of anti-sperm antibodies were developed. For this, sperm surface protein was purified by the immunoaffinity column prepared by the coupling of rabbit anti-human IgG antibodies to Sepharose-4B. Fraction eluted by tris-HCI buffer containing SDS showed a single band having molecular weight of about 60KD on electrophoresis. Enzyme HRP labelled goat anti-human IgG and chemiluminescence aminobutylethyl-isoluminol(ABEI) labelled rabbit anti-human IgG were used for ELISA and CIA, respectively. These two labelled conjugate bound well with human IgG. When serum dilution curves were made to titrate positive serums, two kinds of curves with steep and sluggish slopes were obtained Serum samples were categorized into 3 groups: positive, weak positive and negative based on slope of curve and O.D. values at 1:160 dilution of serum. When ELISA and CIA were compared to conventional method Kibrick test by the determinations of 62 male serums with different diagnosis, the results of ELISA and CIA agreed well, but both disagreed with that of Kibrick test. This study showed that purified sperm surface antigen can be used to develope solid-phase immunoassay systems such as ELISA and CIA which may eliminate the problems encounted the immobilization of living sperm in other tests.

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Comparative Diagnostic Studies on Serologic and Molecular Biological Tests Against Haemorrhagic Fever with Renal Syndrome (신증후출혈열 환자의 혈청학적 및 분자생물학적 진단 검사법 비교)

  • 우영대;문희주;배형준
    • Biomedical Science Letters
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    • v.6 no.2
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    • pp.141-149
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    • 2000
  • The etiologic agents of haemorrhagic fever with renal syndrome (HFRS) in Korea are Hantaan and Seoul virus in the genus Hantavirus, family Bunyaviridae. Antibody titers of sera from HFRS patients against Hantaan virus were measured by immunofluorescent antibody technique (IFAT), enzyme-linked immunosorbent assay (ELISA), high density composite particle agglutination (HDPA) and plaque reduction neutralization test (PRNI). PRNT and nested reverse transcriptase polymerase chain reaction (nested RT-PCR) was used for serotypic differentiation of Hantaviruses against Hantaan and Seoul virus. Eight doubtful HFRS patients showed higher fluorescent, IgG ELISA, agglutination and neutralizing antibody titer by IFAT, ELISA IgG, HDPA and PRNT, respectively Five out of them showed high IgM antibody titer by IgM capture ELISA against Hantaan virus, remarkably. Fifteen HFRS patients showed higher fluorescent antibody titer by IFAT. In PRNT, 12 out of them showed high neutralizing antibody titer against HTNV, 2 against SEOV and 1 against both viruses. In nested RT-PCR using serotype specific-primer, 3 out of them showed positive against HTNV and 1 against SEOV.

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Studies on the Separation of Immunoglobulin and Immunological Response from Korean Native Cattle (한우초유중 Ig의 분리 ${\cdot}$ 정제 및 면역 반응에 관한 연구)

  • Baick, Seung-Cheon;Kim, Yong-Hwi;Shin, Je-Ho;Yu, Je-Hyeon
    • Journal of Dairy Science and Biotechnology
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    • v.15 no.1
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    • pp.1-9
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    • 1997
  • This study was conducted to efficiently separate the Ig from Korean native cattle colostrum and to utilize them as an immunogen for the production of antibodies aginst rabbit. The results obtained were as follows : 1. About 84% of Ig G could be separated from Korean native cattle colostrum by·gel filtration using Superose 12 column on HPLC. The separation profile of Korean native cattle colostral immunoglobulin was similar that of Holstein colostral Ig. 2. Separation of Korean native cattle colostral Ig by anion exchange chromatography using Mono Q column on HPLC was poor resolution chromatographic pattern. 3. Hi-Trap Protein G column showed better results than the Protein A Sepharose CL-4B column in the Ig G binding capacity from Korean native cattle colostral Ig. 4. Protein G Sepharose Fast Flow system resulted in higher Ig g binding capacity as the industrial size scale-up approach. 5. Sufficient titer reaction of antibody to Korean native cattle colostral Ig G was confirmed by ELISA.

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Immune Reaction to Infection by Malassezia pachydercatis in Canine Ezternal Ear Canals (개 외이도내 Malassezia pachydercatis 감염에 대한 면역 반응)

  • 이진희;오태호;한홍율;조상내
    • Journal of Veterinary Clinics
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    • v.13 no.2
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    • pp.130-139
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    • 1996
  • Malassezia pachydercatis균은 개 외이염의 주된 원인균으로 알려져 있으나, 그 감염률은 지역에 따라 많은 차이가 있다. 그러나 그 기병론적인 역할에 대해서는 아직 명확히 밝혀지지 않았다. 이에 본문에서는 국내에서 사육되는 개의 외이염에서 Malassezia pachydercatis 균의 분리율을 조사하고, Malassezia pachydercatis에 대한 혈중 항체와 외이염과의 연관성을 밝혀 Malassezia pachydercatis에 의한 외이염 진단을 위한 도구로서 간접 ELISA 법의 유용성을 평가해 보고자 본 연구를 수행하였다. 총 112두 중, 외이염에 감염된 개 44두와 정상견 68두를 대상으로, 심부외이도에서 취한 가검물을 배양검사하였고, Malassezia pachydercatis 항원에 대한 혈청내 IgG와 IgM 역가를 간접 ELISA법에 의해 측정하였다. 살아있는 Malassezia pachydercatis 균을 3두의 정상견 외이도에 접종하여 Malassezia pachydercatis 균의 실험적 감염에 대한 조직병리학적 소견, 조직내 침투 정도, 그리고 접종 후 시간경과에 따른 IgG와 IgM의 혈중 역가변화를 측정하였다. Malassezia pachydercatis 균은 외이염이 있는 개 귀에서 가장 높게 분리(50.0%)되었고, 정상귀에서는 낮게 분리(6.3%)되었다. Malassezia pachydercatis 항원을 이용한 간접 ELISA 법 진단에서, Malassezia pachydercatis 양성배양인 외이염 개체에서 IgG와 IgM에 대한 민감도는 각각 41.7%, 20.8%이었고 특이도는 88.2%, 85.2% 이었다. 실험적으로 Malassezia pachydercatis 를 접종한 3두중 1두에서만 조직병리학적 변화와 모낭에 Malassezia pachydercatis 균체가 확인되었으며, IgG와 IgM 혈중역가도 유의성 있게 증가하였다. 국내에서 사육되는 개 외이염의 주된 원인균은 Malassezia pachydercatis 균이었고 이 균의 혈중항체는 유의성 있게 증가하였으며(p<0.005), 간접 ELISA법은 Malassezia pachydercatis 에 의한 개외이염의 유용한 진단도구임을 증명하였다.

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A seroepidemiological survey for toxocariasis in apparently healthy residents in Gangwon-do, Korea

  • Park, Hyun-Young;Lee, soo-Ung;Huh, Sun;Kong, Yoon;Magnaval, Jean-Francois
    • Parasites, Hosts and Diseases
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    • v.40 no.3
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    • pp.113-117
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    • 2002
  • We investigated the sero-prevalence of toxocariasis among healthy Korean adults in 1999. A total of 314 sera from normal inhabitants in Whachon-gun, Gangwon do, Korea was examined for specific antibody levels against excretory-secretory products of second stage larvae of Toxocara (TES). The presence of cross-reactions with other helminthiases such as cysticercosis, paragonimiasis, sparganosis or clonorchiasis was also checked by specific IgG ELISA. Sera showing positive reaction against TES were also tested by IgG immunoblot and by IgE ELISA. Out of 314 subjects, 16 was found to be positive by TES IgG ELISA and immunoblot, among whom 12 were also positive by TES IgE ELISA. Among the 16 seropositive samples, two sera showed positive reaction against Paragonimus and sparganum antigen, respectively. These results inferred that cross-reactions were negligible between toxocariasis and other helminthiases. Toxocariasis seroprevalence among Korean rural adults was detected to be approximately 5%.