• Title/Summary/Keyword: porcine rotaviruses

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Characterization of G9 genotype porcine rotavirus isolated in Korea (국내 양돈장에서 분리한 G9형 돼지로타바이러스의 특성)

  • Wang, Jun-Hui;Lee, Seung-Chul;Kang, Shien-Young
    • Korean Journal of Veterinary Service
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    • v.35 no.2
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    • pp.83-90
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    • 2012
  • Porcine rotaviruses are the most common causes of viral gastroenteritis in piglets around the world. The major G genotypes of porcine rotaviruses causing diarrhea were G4, G5 and G11 genotypes. Recently, G9 genotype rotaviruses were problemed at swine farms and frequently recognized from diarrheic piglets. In this study, a porcine rotavirus (PoRV-1) was isolated from piglet showing diarrhea using MA104 cells and confirmed as rotavirus by electron microscopy, genomic RNA electropherotyping and indirect immunofluorescence antibody tests. The nucleotide sequence of the VP7 gene of PoRV-1 was determined and compared with those of other genotype rotavirus strains from other parts of the world. Also, the nucleotide sequences of VP4, VP6 and NSP4 genes of PoRV-1 were determined and compared with those of other rotavirus strains from other countries. The results showed that the PoRV-1 isolate belonged to the G9 genotype and the P, I and E genotypes of PoRV-1 were P[23], I5 and E1, respectively. The Korean G9 PoRV-1 isolate and its nucleotide sequence data would be usefully used for the development of porcine rotavirus vaccines in near future.

Monoclonal antibodies against porcine group C rotavirus VP6 (돼지 group C 로타바이러스 VP6 특이 단클론항체)

  • Yoon, Young-Sim;Lee, Seung-Chul;Woo, Sang-Kyu;Cho, Kyoung-Oh;Kang, Shien-Young
    • Korean Journal of Veterinary Service
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    • v.35 no.3
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    • pp.175-182
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    • 2012
  • Rotaviruses have been known to be a major etiological agent of gastroenteritis in both infants and young animals. Subsequently new rotaviruses, which were morphologically indistinguishable but antigenically and electrophoretically distinct with each other, were reported from several animals throughout world including Korea. These new rotaviruses were named as non-group A or group B or group C rotaviruses and so on. It has been very difficult to isolate and grow the non-group A rotaviruses in vitro, and this has greatly limited the characterizations of non-group A rotaviruses and serological studies. In this study, monoclonal antibodies (MAbs) against porcine non-group A rotavirus were produced and characterized. The VP6 gene of porcine group C rotavirus Korean isolate(#06-52-1) was cloned and expressed. For expression of VP6 gene, baculovirus expression system was applied. The VP6 gene and expressed protein in the recombinant virus were confirmed by polymerase chain reaction (PCR), indirect fluorescence antibody (IFA) test and Western blot, respectively. The expressed VP6 was used for MAbs production. The MAbs produced in this study would be promising as diagnostic reagents for detection of group C rotavirus infection.

Studies on the production and characterization of monoclonal antibodies against bovine rotaviruses isolated in Korea (소 로타바이러스(국내분리주)에 대한 단크론항체 생산 및 특성에 관한 연구)

  • Ahn, Jae-moon;Cho, Sun-hee;Kang, Shien-young
    • Korean Journal of Veterinary Research
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    • v.36 no.2
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    • pp.395-403
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    • 1996
  • Monoclonal antibodies(MAbs) against field isolates of the bovine rotavirus A strain(G6), V strain(G10) and reference I-801 strain(G8) were produced and characterized. Six MAbs(4C2, 4D9, 5E1, 5E7, 5D5, 3E4) against A strain had neutralizing activity and reacted only with the G6 bovine rotaviruses determined by fluorescence focus neutralization (FFN) test. Otherwise, five neutralizing MAbs(1G2, 2G6, 5E2, 5E12, 5H7) against I-801 strain neutralized the G6 and G8 bovine rotaviruses. Five non-neutralizing MAbs(5F12, 7F12, 5E11, 2A11, 2B12) were VP6-specific and cross-reacted with all bovine and porcine rotaviruses examined by fluorescence antibody(FA) test. None of the MAbs reacted with bovie viral diarrhea virus(BVDV) and bovine coronavirus(BCV) determined by FA and FFN test.

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Inhibitory Effect on Rotavirus by Exopolysaccharides Extracted from Kefir (Kefir에서 추출한 Exopolysaccharide의 Rotavirus의 저해효과)

  • Song, Jin-Ook;Kim, Tae-Jin;Kim, Yong-Hui
    • Food Science of Animal Resources
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    • v.27 no.4
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    • pp.538-542
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    • 2007
  • This study was carried out to investigate the toxicity of exopolysaccharides (EPS) from kefir toward MA104 cells and evaluate the inhibitory effects of kefir EPS on rotavirus infection. The results obtained are summarized as follows: Lactic acid bacteria (Lactobacillus fermentum, L. acidophilus, L. brevis) and yeasts (Candida kefyr, Cryptococcus albidus, Pichia ohmeri) were isolated and identified from kefir grain and culture. At 1% EPS, the inhibitory effects of EPS on the infection of MA-104 cells using the MTT assay were $72.52{\pm}6.48%$ for human rotavirus (KU), $36.06{\pm}7.63%$ for bovine rotavirus (NCDV), and $81.66{\pm}1.11%$ for porcine rotavirus (OSU). At 1/128% EPS, the effects were $24.98{\pm}4.58%$ for human rotavirus (KU), $4.71{\pm}6.16%$ for bovine rotavirus (NCDV), and $4.05{\pm}14.90%$ forporcine rotavirus (OSU). EPS isolated from kefir have inhibitory effects on rotaviruses of various serotypes and rotaviruses from different animals.

Studies on the VP4 and VP7 Genes of Bovine Rotaviruses from Field Samples Using RT-PCR and RFLP Analysis (RT-PCR과 RFLP법을 이용한 국내 소 로타바이러스 VP4 및 VP7 유전자의 특성 규명)

  • Jeon, Seong-Jin;Chang, Chung-Ho;Chung, Chung-Won;Kim, Won-Yong;Kang, Shien-Young
    • The Journal of Korean Society of Virology
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    • v.28 no.2
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    • pp.165-174
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    • 1998
  • Characterizations of the VP4 (P type) and VP7 (G type) genes of Korean isolates of bovine rotavirus were performed using RT-PCR/RFLP and nucleotide sequencing analysis. After RT-PCR amplification of partial length (1094bp) of the VP4 and full length (1062bp) of the VP7 genes, amplified PCR products were digested with restriction endonucleases and digestion patterns were compared with those of reference rotaviruses. With the VP4 genes, four RFLP (A-D) profiles were observed; three (A, Band C) were the same as those of bovine rotavirus NCDV (P[1]), IND (P[5]) and B223 (P[11]), respectively. Profile D was the same as that of porcine rotavirus OSU (P[7]). With the VP7 genes, five RFLP profiles (I-V) were observed; three of them (I, II and III) were the same as those of bovine rotavirus NCDV (G6), Cody 1-801 (G8), and B223 (G10), respectively. Profile IV and V were atypical to those of reference bovine rotaviruses used in this study. These two profiles were identified as G6 and G5, respectively, after analyzing and comparing the nucleotide sequences. The G typing analysis revealed that 61.9% (26/42) were G6, which included G6 subtype; 28.6% (12/42) were G5; 7.1% (3/42) were G10; 2.4% (1/42) were G8. The P typing analysis revealed that 54.8% (23/42) were P[5]; 28.6% (12/42) were P[7]; 11.8% (5/42) were [11]; 4.8% (2/42) were P[1]. Our results showed that G6/P[5] were the most prevalent rotaviruses in diarrheic calves in Korea. Also, this is the first report that G5/P[7] rotaviruses were identified from cattle with diarrhea.

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Detection of bovine rotavirus antigen by enzyme linked immunosorbent assay (효소면역법에 의한 소 로타바이러스 항원 검출)

  • 안재문;유기조;이용희;이종인
    • Korean Journal of Veterinary Service
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    • v.19 no.1
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    • pp.30-38
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    • 1996
  • Enzyme-linked immunosorbent assay (ELISA) was developed to detect rotavirus antigen in fecal samples using VP6-specific monoclonal antibody(2B12). The ELISA for rotavirus antigen detection found to have specificity to all bovine and porcine rotaviruses tested but not to bovine viral diarrhea virus and bovine coronavirus. The ELISA appeared to have similar sensitivity and specificity compared to fluorescence antibody assay(FA) and electropherotyping (PAGE).

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RT-PCR for detection of group A, B and C porcine rotaviruses (RT-PCR 기법을 이용한 돼지 로타바이러스 주요 혈청법 감별진단)

  • Park, Choi-kyu;Lyoo, Young-soo;Paul, Prem S.
    • Korean Journal of Veterinary Research
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    • v.38 no.1
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    • pp.85-89
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    • 1998
  • 본 연구에서는 돼지 로타바이러스의 혈청형 A, B 및 C를 동시에 진단할 수 있는 RT-PCR 기법을 개발하였다. 각각의 혈청형에 특이적인 primer를 이용하여 RT-PCR 기법으로 분변시료에서 직접 바이러스 동정을 실시한 결과, 23건의 로타바이러스 감염 분변 시료에서 13건이 혈청형 A, 3건이 혈청형 B, 2건이 혈청형 C로 나타나 국내에서도 A, B 및 C형의 돼지 로타바이러스가 공히 발생하고 있음을 확인하였으며, 발생분포는 외국의 예와 유사하였다. 이 RT-PCR기법은 돼지 로타바이러스의 주요 혈청형인 A, B 및 C형의 동시감별진단법으로 이용될 수 있을 것으로 판단된다.

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Nucleotide Sequence Homology in Rotaviruses (Rotaviruses의 염기배열 유사성 측정)

  • ;Spendlove, Rex S.;Barnett, Bill B
    • Korean Journal of Microbiology
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    • v.26 no.3
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    • pp.155-161
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    • 1988
  • Nucleotide sequence homology between bovine, simian, and porcine rotavirus was determined by the RNA:RNA hybridization technique. Single stranded RNA, prepared in vitro with EDTA activated endogeneous viral RNA polymerase, was hhbridized with tritium labeled bovine rotavirus genomic RNA. The heteroduplex RNA was treated with single stranded RNA specific ribonucleases and the RNase resistant hybrid RNA was precipitated, and collected by filtration on a filter paper. Seventy four percent RNA sequence homology between bovine and simian rotavirus and 8 percent RNA sequence homology between bovine and porcine rotavirus was confirmed by hybridization between tritium labeled single stranded RNA and viral genomic RNA.

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Development of diagnostic methods for rotavirus from pigs using monoclonal antibody (단크론 항체를 이용한 돼지 로타바이러스의 진단법 개발)

  • Kang, Shien-young
    • Korean Journal of Veterinary Research
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    • v.32 no.4
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    • pp.569-577
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    • 1992
  • Group A 로타바이러스 VP6에 특이적으로 반응하는 단크론 항체를 이용하여 로타바이러스 감염이 의심되는 돼지 분변으로부터 로타 바이러스를 검색할 수 있는 효소면역측정법을 개발하였다. 이 효소면역측정법에서는 capture antibody로서 protein A-sepharose를 이용하여 단크론 항체로부터 순수 분리한 immunoglobulin을 사용하였으며 detecting antibody는 토끼 면역혈청으로부터 순수 분리한 immunoglobulin에 biotin을 label하여 사용하였다. 개발된 효소면역측정법의 민감도와 특이성을 전자현미경법 및 형광항체법의 것과 비교하여 보았을 때 서로 유사하였으며 분변재료로부터 로타바이러스를 검색하는데 유용한 것으로 나타났다. 개발된 효소면역측정법은 야외로부터 로타바이러스 검색을 위하여 수집된 많은 양의 분변재료를 실험실내에서 screen하는데 유용하게 사용될 것으로 생각된다.

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A comparative study on immunogenicity of the porcine epidemic diarrhea virus live-vaccine and inactivated-vaccine (돼지유행성설사병(PEDV) 생독과 사독백신의 면역형성 비교연구)

  • Kwon, Mee-Soon;Cho, Hyun-Ung;Lee, Eun-Mi;Lee, Ji-Yoog;Seo, Heyng-Seok;Im, Jeong-Cheol;Hur, Boo-Hong
    • Korean Journal of Veterinary Service
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    • v.32 no.3
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    • pp.201-207
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    • 2009
  • Porcine transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV) and rotaviruses are considered as the most important causative agents of diarrhea in piglets. The study established 3 method vaccination programs to prevent PEDV. A (LL)group inoculated twice vaccinations on 2-weeks-interval during the late term of pregnant sows with PEDV live vaccine. The B (LKK) group was applied that one time single PEDV live vaccine at the pre-mate followed by the TGEV PEDV combined inactivated vaccine (twice vaccination on 2-weeks interval at the third-trimester). C (KK) group was applied to sow which inoculated twice vaccination on 2-weeks-interval during the late term of pregnant sows with by the TGEV, PEDV combined inactivated vaccine. As the result of SN test on sows in the pig farm before vaccination, antibody titers was showed 9/45 (20.0%). By comparison with the serum neutralizing antibody titers against PEDV of the vaccination programs after PEDV of the vaccination, A group and B group vaccination method was higher than those of C group in sows. In the piglets up to 2 weeks of age, A group was showed antibody titers of 17/22 (81.8%) that showed 2-128, and B group was showed antibody titers of 30/37 (81.1%) that showed 2-512, and C group was showed antibody titers of 14/28 (50.0%) that showed 2-32. On the other hand, PEDV antibody titers were tested for the survey. As the results of SN test, Aujeszky's disease survey in 54 pig farms from november 2005 to august 2006, antibody titers of 47/286 (16.4%) showed above 2. Five breeding farms were antibody titers of 38/77 (49.4%), Wanggung zone farms antibody titers of 59/85 (69.4%). In pigs farms vaccinated the first of twice PEDV live vaccine, and after 6 month, the second of twice TGEV PEDV combined inactivated vaccine (LLKK, 256-1024 titer) method was higher than those of vaccinated twice the early term of pregnant, and twice the late term of pregnant sows of PEDV live vaccine (LLLL, 32 titer).