• 제목/요약/키워드: Neutralization assay

검색결과 59건 처리시간 0.023초

Rabies neutralizing antibody titers in Korean dogs and cats intended for overseas travel

  • Dong-Kun Yang;Yu-Ri Park;Ha-Hyun Kim;Eun-Ju Kim;Hye Jeong Lee;Young-Ju Cheong;Bang-Hun Hyun
    • 대한수의학회지
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    • 제63권2호
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    • pp.13.1-13.6
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    • 2023
  • Animals imported from abroad are a cause of rabies outbreaks in many countries. Therefore, rabies serology testing for dogs and cats traveling abroad is an important measure to reduce the incidence of rabies. Rabies virus antibodies were measured in sera collected from 2,367 dogs and 894 cats between 2017 and 2021. A serum sample with a value of 0.5 IU/mL or higher was considered a pass. The overall pass rates for rabies virus were 96.4% in dogs and 98.4% in cats. The mean rabies virus neutralization assay titers were higher in cats than in dogs and in female than in male animals. According to age, 6-year-old dogs and 9-year-old cats had the highest virus neutralization assay titers. Of the failure cases, 53.0% (53/100) were dogs or cats less than 1 year old. Although the average failure rates in dogs and cats were low at 3.5% and 1.6%, respectively, the factors influencing failure were age and vaccine manufacturer. Therefore, it is necessary to observe the vaccination interval and timing of blood collection after boosting.

Detection of immunity in sheep following anti-rabies vaccination

  • Hasanthi Rathnadiwakara;Mangala Gunatilake;Florence Cliquet;Marine Wasniewski;Mayuri Thammitiyagodage;Ramani Karunakaran;Jean-Christophe Thibault;Mohamed Ijas
    • Clinical and Experimental Vaccine Research
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    • 제12권2호
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    • pp.97-106
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    • 2023
  • Purpose: Rabies is a fatal but preventable disease with proper pre-exposure anti-rabies vaccination (ARV). Dogs, as household pets and strays, are the reservoir and vector of the disease, and dog bites have been associated with human rabies cases in Sri Lanka over the past few years. However, other susceptible species having frequent contact with humans may be a source of infection. One such species is sheep and immunity following ARV has never been tested in sheep reared in Sri Lanka. Materials and Methods: We have tested serum samples from sheep reared in the Animal Centre, Medical Research Institute of Sri Lanka for the presence of anti-rabies antibodies following ARV. Sheep serum samples were tested with Bio-Pro Rabies enzyme-linked immunosorbent assay (ELISA) antibody kits used for the first time in Sri Lanka and our results were verified by a seroneutralization method on cells (fluorescent antibody virus neutralization, FAVN test) currently recommended by World Organization for Animal Health and World Health Organization. Results: Sheep received annual ARV and maintained high neutralizing antibody titers in their serum. No maternal antibodies were detected in lamb around 6 months of age. Agreement between the ELISA and FAVN test, i.e., coefficient concordance was 83.87%. Conclusion: Annual vaccination in sheep has an effect on maintaining adequate protection against rabies by measurements of anti-rabies antibody response. Lambs need to be vaccinated earlier than 6 months of age to achieve protective levels of neutralizing antibodies in their serum. Introducing this ELISA in Sri Lanka will be a good opportunity to determine the level of anti-rabies antibodies in animal serum samples.

A rapid and quantitative fluorescent microsphere immunochromatographic strip test for detection of antibodies to porcine reproductive and respiratory syndrome virus

  • Wei, Yanqiu;Yang, Baozhi;Li, Yunlong;Duan, Yongcheng;Tian, Deyu;He, Baoxiang;Chen, Chuangfu;Liu, Wenjun;Yang, Limin
    • Journal of Veterinary Science
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    • 제21권4호
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    • pp.68.1-68.8
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    • 2020
  • A fluorescent microsphere-based immunochromatographic strip test (FICT) was developed for the rapid, sensitive, and quantitative detection of porcine reproductive and respiratory syndrome virus (PRRSV) antibodies at the pen-side. The assay was based on the formation of a sandwich immune-complex (anti-pig IgG-PRRSV antibodies-NSP7/N), which was validated by a comparison with IDEXX-ELISA using 3325 clinical specimens. The diagnostic specificity, sensitivity, and accuracy of FICT were 97.28, 93.41, and 94.95%, respectively. FICT showed a good correlation with the virus neutralization assay. Overall, a promising pen-side diagnostic tool was developed for the rapid and quantitative detection of PRRSV antibodies within 15 min.

Optimal Conditions of Single Cell Gel Electrophoresis (Comet) Assay to detect DNA single strand breaks in Mouse Lymphoma L5178Y cells

  • Ryu, Jae-Chun;Kwon, Oh-Seung;Kim, Hyung-Tae
    • 한국환경성돌연변이발암원학회지
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    • 제21권2호
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    • pp.89-94
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    • 2001
  • Recently, single cell gel electrophoresis, also known as comet assay, is widely used for the detection and measurement of DNA strand breaks in vitro and in vivo in many toxicological fields such as radiation exposure, human monitoring and toxicity evaluation. As well defined, comet assay is a sensitive, rapid and visual method for the detection of DNA strand breaks in individual cells. Briefly, a small number of damaged cells suspended in a thin agarose gel on a microscope slide were lysed, unwinded, electrophoresed, and stained with a fluorescent DNA binding dye. The electric current pulled the charged DNA from the nucleus such that relaxed and broken DNA fragments migrated further. The resulting images which were subsequently named for their appearance as comets, were measured to determine the extent of DNA damages. However, some variations could be occurred in procedures, laboratories's conditions and kind of cells used. Hence, to overcome and to harmonize these matters in comet assay, International Workshop on Genotoxicity Test Procedure (IWGTP) was held with several topics including comet assay at Washington D.C. on March, 1999. In spite of some consensus in procedures and conditions in IWGTP, there are some problems still remained to be solved. In this respect, we attempted to set the practical optimal conditions in the experimental procedures such as lysis, unwinding, electrophoresis and neutralization conditions and so on. First of all, we determined optimal lysis and unwinding time by using 150 $\mu$M methyl methanesulfonate (MMS) which is usually used concentration. And then, we determined optimal positive control concentrations of benzo(a)pyrene (BaP) and MMS in the presence and absence of S9 metabolic activation system, respectively.

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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
    • 한국미생물·생명공학회지
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    • 제46권3호
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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.

전염성 F낭병에 대한 혈청학적 연구 (The serological studies on infectious bursal disease)

  • 정영미;서석열;도홍기;조정곤;노수일
    • 한국동물위생학회지
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    • 제23권3호
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    • pp.271-279
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    • 2000
  • This study was carried out to provide the fundamental information for development of proper vaccination program against infectious bursal disease(IBD) to the local chicken farms. The antigen detection was peformed from 8 samples of bursa of Fabricius with agar gel precipitation(AGP) and indirect immunofluorescent assay(IFA), And also, the antibodies in serum samples were detected by the various serological methods such as commercial ELISA assay, AGP and virus neutralization(VN) test. 1. The antigen detection rates were 25% for AGP which is 2 out of 8 farms and 10 out of 40 bursas, and 25% which Is 2 out of 8 farms and 20% 8 out of 40 bursas for IFA, respectively. 2. The mean titer of maternal antibody (>3,000) existed until 10 days of the age with ELISA-GMT. 3. The antibody positive rates which are over 80% showed until 5 days of the age with ELISA and at 10 days of the age with AGP except one, but none of them showed from 1 day of the age. This report came to conclusions that both the protective maternal antibody titers and the antigen positive rates were significant until at the 10 days of the age.

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하리자돈(下痢子豚) 분변(糞便)에서 Rotavirus 분리(分離) 동정(同定) (Isolation and Identification of Porcine Rotavirus from Piglets with Diarrhoea)

  • 김희선;최정옥;김두희;유영수;안수환
    • 대한수의학회지
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    • 제28권1호
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    • pp.89-97
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    • 1988
  • Twenty-eight porcine rotavirus were isolated from piglets with diarrhoea in chonnam province. According to the age, 41 to 60 day old pigs showed the highest isolation frequency among the post weaning pigs. The characteristics of the field isolates were determined by electronmicroscopy(EM), immunofluorescent assay(FA), and electrophoretic migration patterns of the genome profiles. Some of the isolates showed remarkable haemagglutination activity against rabbit and dog erythrocytes, ranged from 4 to 2848, respectively. At least 3 serotypes of porcine rotavirus were recognized by serum neutralization test using serotype specific rotavirus hyperimmune sera.

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Identification of a mimotope of an infectious bronchitis virus S1 protein

  • Zhou, Jingming;Li, Jianan;Li, Yanghui;Liu, Hongliang;Qi, Yanhua;Wang, Aiping
    • Journal of Veterinary Science
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    • 제22권4호
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    • pp.49.1-49.6
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    • 2021
  • The S1 protein of the infectious bronchitis virus (IBV) is a major structural protein that induces the production of the virus-neutralization antibodies. The monoclonal antibody against the IBV M41 S1 protein was used as a target for biopanning. After three rounds of biopanning, randomly selected phages bound to the monoclonal antibody. Sequence analysis showed that the dominant sequence was SFYDFEMQGFFI. Indirect competitive enzyme-linked immunosorbent assay showed that SFYDFEMQGFFI is a mimotope of the S1 protein that was predicted by PepSurf. The mimotope may provide information for further structural and functional analyses of the S1 protein.

Prevalence and genotypes of pestivirus in Korean goats

  • Yang, Dong-Kun;Kweon, Chang-Hee;Kim, Byoung-Han;Choi, Cheong-Up;Kang, Mun-Il;Hyun, Bang-Hun;Hwang, In-Jin;Lee, Cheong-San;Cho, Kyoung-Oh
    • 대한수의학회지
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    • 제48권1호
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    • pp.83-88
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    • 2008
  • In total, 1,142 serum samples were collected from 223 goat flocks rising in five different regions of Korea. These samples were screened for the presence of border disease virus (BDV) antibodies using an enzyme linked immunosorbent assay. Of the 1,142 samples, we found 47 bovine viral diarrhea virus (BVDV) positive cases (4.1%). These positive serum samples were also examined further by using the virus neutralization test against BDV. In addition, samples were tested for both BVDV and classical swine fever virus (CSFV). All of the samples that were seropositive for BDV also demonstrated positive antibody titers against BVDV and CSFV. Due to their common antigenicity, we also determined further the prevalence and carried out virus neutralization test against three pestiviruses: 314 of the goat samples were screened using reverse transcription polymerase chain reaction with primer pairs specific to common pestivirus genome regions. Overall, 1.6% (5/314) of the samples tested was positive for pestivirus. Based on the nucleotide sequence data and the phylogenetic analysis, three isolates were characterized as BVDV type 1 and two isolates as BVDV type 2. However, none of the isolates could be classified as BDV. These results indicate that BVDV-1 and BVDV-2 are the pestivirus strains circulating among Korean goat populations.