• 제목/요약/키워드: PCR-ELISA

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경북지역 산란계에서 Avian pneumovirus에 대한 항체가 및 바이러스 검출 조사 (Examination of seroprevalence and detection of Avian pneumovirus from layer hens in Gyeongbuk province)

  • 김정은;황지영;배동록;성명숙;김순태;김상윤
    • 한국동물위생학회지
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    • 제30권1호
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    • pp.43-49
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    • 2007
  • Avian pneumovirus(APV), also known as avian rhinotracheitis(ARTV), affects both turkeys and chickens and is known to be the primary causative agent of turkey rhinotracheitis(TRT). The aim of this study was to establish the presence or absence of antibodies to APV by an enzyme-linked immunosorbent assay(ELISA) and confirm APV by a reverse transcriptase-polymerase chain reaction(RT-PCR). The tested serum and feces were collected from laying hens in Gyeongbuk province. The positive farms with antibody against APV by ELISA were 90(96.7%) of 93 and positive serum samples were 433(93.1%) of 465 different sera. By regional group, sera from Uiseong, Cheongsong and Bonghwa were noted as 100% positive and positive rates of samples from Yeongju, Andong and Yeongyang were 93.3%, 85.7% and 50%, respectively. However, APV was not detected in feces samples by RT-PCR.

서부경남 재래시장을 대상으로한 유전자 재조합 대두의 모니터링 (Monitoring of Genetical Modified Soybean Sold at Local Open Market in Western Gyeongnam)

  • 심원보;남백상;최주미;정순천;정덕화
    • 한국환경보건학회지
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    • 제32권2호
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    • pp.126-131
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    • 2006
  • The objective of this study was monitoring of genetically modified soybean by PCR and ELISA. We collected 60 soybean samples from the open markets located in Western Gyeongnam (Sacheon, Hamyang, Hadong, Sanchung, Uiryung, Geochang, and Hapcheon). A total of 60 soybeans was examined and 14 genetical modified soybean (GMS) were detected by PCR. The GMS rate of selling soybean in Uiryung, Hadong, Sacheon, and Hapcheon was 50.0%, 37.5%, 33.3% and 25%, respectively. The 7 of 14 GMSs were positive by ELISA and most of positive samples were below 3% GMS but 1 (Uiryung 1) of the positive samples was over the 3% which is maximum permit limit in Korea. These results mean that merchants of open market did not express for selling soybean mixed with GMS, so consumers did not recognize GMO. Therefore, we thought that education of GMO for merchant of open market need to recognize about GMO maximum permit limit.

경북지역 돼지의 바이러스성 유사산 원인조사 (Etiological Study of Porcine Viral Abortions and Stillbirths in Gyeongbuk Province)

  • 채태철;김성국;조광현;어경연;권오덕
    • 한국임상수의학회지
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    • 제30권4호
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    • pp.236-240
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    • 2013
  • A total of 170 litters (575 samples) of aborted and stillbirth fetuses submitted to the Gyeongsangbuk-Do Veterinary Service Laboratory (GVSL) between January 2006 and December 2010 from pig farms in Gyeongbuk province were studied to identify porcine abortion- and stillbirth-associated viruses such as Porcine parvovirus (PPV), Encephalomyocarditis Virus (EMCV), Japanese Encephalitis Virus (JEV), Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), and Aujeszky's Disease Virus (ADV). Virus was not detected by PCR in 36 litters, but viral antibody was detected by HI and ELISA in 93 litters. The majority of etiological viruses were PPV (67 litters, 39.4%), EMCV (50 litters, 29.4%), PRRSV (15 litters, 8.8%), and JEV (11 litters, 6.5%); ADV was not detected by either PCR or ELISA. Single infection occurred in 52 litters (30.6%), co-infection occurred in 41 litters (24.1%), and unknown cases with no detection of any of the five viruses occurred in 77 litters (45.3%).

Genetic Diversity in the Coat Protein Genes of Prune dwarf virus Isolates from Sweet Cherry Growing in Turkey

  • Ozturk, Yusuf;Cevik, Bayram
    • The Plant Pathology Journal
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    • 제31권1호
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    • pp.41-49
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    • 2015
  • Sweet cherry is an important fruit crop with increasing economical value in Turkey and the world. A number of viruses cause diseases and economical losses in sweet cherry. Prune dwarf virus (PDV), is one of the most common viruses of stone fruits including sweet cherry in the world. In this study, PDV was detected from 316 of 521 sweet cherry samples collected from 142 orchards in 10 districts of Isparta province of Turkey by double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA). The presence of PDV in ELISA positive samples was confirmed in 37 isolates by reverse transcription- polymerase chain reaction (RT-PCR) method. A genomic region of 862 bp containing the coat protein (CP) gene of PDV was re-amplified from 21 selected isolates by RT-PCR. Amplified DNA fragments of these isolates were purified and sequenced for molecular characterization and determining genetic diversity of PDV. Sequence comparisons showed 84-99% to 81-100% sequence identity at nucleotide and amino acid level, respectively, of the CP genes of PDV isolates from Isparta and other parts of the world. Phylogenetic analyses of the CP genes of PDV isolates from different geographical origins and diverse hosts revealed that PDV isolates formed different phylogenetic groups. While isolates were not grouped solely based on their geographical origins or hosts, some association between phylogenetic groups and geographical origins or hosts were observed.

전염성 F낭병 항체가 조사 및 유전자 분석 (Seroprevalence of infectious bursal disease (IBD) and genetic sequence analysis of IBD virus)

  • 강미선;추금숙;조현웅;한재철
    • 한국동물위생학회지
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    • 제35권2호
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    • pp.91-97
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    • 2012
  • The strategy for infectious bursal disease (IBD) control and its success rate under field conditions depends on hygiene management, IBD field pressure, level, and variation in maternally derived IBD antibodies. This study investigated the level of IBD-specific antibody by ELISA and the prevalence of IBD virus by PCR in broilers, white-semi broilers, and Korean native chickens raised in Jeongeup, Jeonbuk. IBD-specific maternally derived antibodies were measured from 698 chickens and the mean titers of maternal antibodies were $3,572{\pm}1,402$ in broilers, $1,262{\pm}762$ in white-semi broilers, and $1,932{\pm}912$ in Korean native chickens. At 2 weeks after vaccination, the geometric mean antibody titers of broiler, white-semi broiler, and Korean native chicken were $582{\pm}427$, $3255{\pm}1,080$, and $1,023{\pm}499$, respectively. According to sequence analysis of the variable virion protein 2 gene, 4 isolates were found to be very virulent IBDV, 9 isolates classical virulent, and 2 isolates intermediate plus vaccine strain.

토마토반점위조바이러스(TSWV) 저항성 토마토 유전자원 탐색 (Screening of Tomato Spotted Wilt Virus Resistance in Tomato Accessions)

  • 한정헌;최학순;이준대;김재덕;이원필;최홍수;김정수;윤재복
    • 원예과학기술지
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    • 제30권2호
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    • pp.171-177
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    • 2012
  • Sw5-2 SCAR 분자표지와 생물검정법을 이용하여 토마토 유전자원 94종의 $Tomato$ $spotted$ $wilt$ $virus$(TSWV) 저항성을 조사하였다. Sw5-2 SCAR 분자표지의 PCR 산물은 대략 574bp, 500bp, 462bp였는데, 크기가 가장 큰 PCR 산물이 Sw5-b 저항성 대립유전자와 연관되어 있었다. Sw5-b 저항성 대립유전자는 3개 수집종('Eureta', 10-318, 10-321)에서 관찰되었는데, 접종한 개체 가운데 이들 가운데 일부는 TSWV-pb1(토마토 분리주)에 일시적으로 감염되어 회복되거나 줄기에 괴사 병징을 보였다. ELISA 검사에서 음성으로 판명된 수집종 당 1개체씩 총 35개체를 선발하여 병징 발현 및 바이러스 감염 유무를 추가로 조사하였다. 접종 5개월 이후에 병징이 나타나지 않은 26개체를 대상으로 RT-PCR을 이용하여 TSWV 감염유무를 조사한 결과, 모든 개체에서 TSWV의 RT-PCR 산물이 약하게 증폭되었고, 이들 PCR 산물의 증폭 수준은 'Eureta'와 비슷하였다. 선발된 유전자원의 저항성은 조직 내 TSWV의 농도를 낮게 하는데 중요한 역할을 하고 이들은 Sw5를 포함한 여러 가지 유전자들에 의해 양적으로 조절되는 것으로 판단된다.

인간의 단핵구와 비만세포에서 다양한 아토피 유발물질이 사이토카인 유전자의 발현에 미치는 영향 (Effects of atopic dermatitis induced materials on the expression of cytokine genes in human monocytes and mast cells)

  • 박경숙;김경준
    • 한방안이비인후피부과학회지
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    • 제23권2호
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    • pp.41-56
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    • 2010
  • Objectives : On an experimental basis, the effects of atopic dermatitis induced materials on the expression of cytokine genes in human monocytes (THP-1, U937) and mast cells were studied. This study was carried out to be considered a fundamental knowledge in the research on the good of oriental medicine. Methods : After culturing THP-1, U937, and HMC-1, with the three different concentrations of LPS ($1\;{\mu}g/ml$), DPE ($10\;{\mu}g/ml$), and DNCB ($1\;{\mu}g/ml$), atopic dermatitis induced materials were treated in the culture medium. To investigate cytokine genes expression patterns, with lysis buffer and separation reagent, total RNA was extracted from THP-1, U937, and HMC-1 at intervals of 0, 12, 24, and 48 hours. Both cytokine mRNA expression patterns by atopic dermatitis induced materials and change of cytokine genes expression patterns in relation to atopy by selenium were analyzed with RT-PCR. Also IL-4 and INF-$\gamma$, which were secreted in the HMC-1, were analyzed using ELISA method. Results : 1. After treating THP-1 and U937 with LPS, DPE, and DNCB, there was no significant change in cytokine genes themselves, but various cytokines (IL-4, IL-6, IL-8, IL-13, IFN-$\gamma$, IFN-a, MCP-1, B2-MG) were expressed. 2. In the case of HMC-1, the expressions of IL-6 and IL-8 were significantly increased in the analysis of mRNA expression by dust mite allergens in DPE. 3. As a result of ELISA method, it is certain that IL-4 and IFN-$\gamma$ protein were secreted in the HMC-1 by DPE. 4. Selenium, an essential trace element, decreased the IL-10 and IL-13 expression in the HMC-1 by DPE. Conclusion : The results suggest that it is necessary to choose proper atopic dermatitis induced materials and suitable cultured cells in establishment of in vitro model of atopic dermatitis.

Agrobacterium을 이용한 형질전환 상추의 세포 현탁배양으로부터 hGM-CSF의 생산 (Production of hGM-CSF from Cell Suspension Culture of Transformed Lettuce Using Agrobacterium-mediated Transformation System)

  • 김영숙;김미영;권태호;양문식
    • Journal of Plant Biotechnology
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    • 제30권1호
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    • pp.97-102
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    • 2003
  • hGM-CSF가 식물세포 현탁 배양을 통하여 생산이 가능한지를 조사하기 위하여 hGM-CSF를 포함하고 있는 A. tumerfaciens LBA4404를 가지고 상추에 형질전환시켰다. 형질전환된 상추로부터 캘러스를 유도하여 캘러스를 이용한 세포배양체계를 확립하였다. PCR과 Southern blot analysis 결과 상추에 hGM-CSF 유전자가 도입된 것을 확인하였으며, Northern blot analysis 결과 상추식물체에 hGM-CSF 유전자가 발현됨을 확인하였다. 현탁 배양 세포로부터 분비된 hGM-CSF를 ELISA를 이용하여 측정한 결과 149.0 $\mu\textrm{g}$/L가 생산됨 을 확인하였다 이러한 결과는 상추의 현탁 배양 세포가 hGM-CSF와 같은 치료용 단백질의 생산 숙주로 이용될 수 있음을 보여주었다.

가는갯능쟁이(Atriplex gmelinii) 추출물과 용매분획물의 MMP-2와 MMP-9 활성 저해효과 (Inhibition of MMP-2 and -9 by Crude Extracts and Their Solvent-partitioned Fractions from the Halophyte Atriplex gmelinii)

  • 박민정;김준세;공창숙;서영완
    • Ocean and Polar Research
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    • 제41권2호
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    • pp.79-88
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    • 2019
  • In this study, the inhibitory effect of Atriplex gmelinii C. A. Mey. against the activity of MMP-2 and MMP-9 secreted from phorbol-12-myristate-13-acetate (PMA)-stimulated HT-1080 cells was evaluated by gelatin zymography and enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase-chain reaction (RT-PCR), and Western blot assay. Specimens of the halophyte A. gmelinii were extracted twice for 24 hr with methylene chloride ($CH_2Cl_2$), and then twice with methanol (MeOH), in turn. Each extract significantly inhibited the enzymatic activities in gelatin zymography and MMP ELISA kit, and expression of MMP-2 and 9 in mRNA and protein levels. Two crude extracts were combined and then the combined crude extracts were fractionated into n-hexane, 85% aqueous methanol (85% aq.MeOH), n-butanol (n-BuOH), and water ($H_2O$) fractions, according to solvent polarity. Among solvent-partitioned fractions, the 85% aq.MeOH fraction showed the strongest inhibitory effect against MMP-2 and -9 in gelatin zymography and MMP ELISA kit. In RT-PCR, all solvent-partitioned fractions significantly suppressed mRNA expression of MMP-2 and -9. On the other hand, in Western blot assay, all solvent-partitioned fractions except $H_2O$ significantly reduced expression levels of protein. HT 1080 cell migration was most significantly inhibited by the n-BuOH fraction followed by the 85% aq.MeOH and $H_2O$ fractions. These results suggest that A. gmelinii could be used as a potential source to inhibit tumor cell metastasis.

Update on Distribution and Genetic Variability of Plum pox virus Strains in Bulgaria

  • Kamenova, Ivanka;Borisova, Anelija
    • The Plant Pathology Journal
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    • 제35권3호
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    • pp.243-256
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    • 2019
  • Field surveys for Plum pox virus (PPV) infection were conducted in stone fruit orchards all over Bulgaria. In total, 1168 out of 3020 leaf samples from cultivated Prunus spp. and wildly growing P. cerasifera trees reacted positive for PPV in DASI-ELISA with the universal monoclonal antibody (MAb) 5B. Further ELISA analyses showed that 987 and 127 isolates belonged to PPV-M and PPV-D serotypes, respectively. The plum and P. cerasifera showed 82.0% and 50.5% levels of infection, respectively followed by the peach (40.0%) and the apricot (32.0%). Five hundred fifty one PPV isolates were further typed by IC-RT-PCR with PPV-Rec, -M and -D-specific primers, targeting (Cter)NIb-(Nter) CP genome region, as 125 isolates were sequenced. The results revealed the presence of PPV-Rec, PPV-M and PPV-D and mixed infections of these strains. PPV-Rec was the most prevalent strain (49.0%), followed by PPV-M (40.1%), while PPV-D was the less spread strain (8.2%). PPV-Rec was the most common strain in plums, including the eight "old-aged" trees from the region of the first Sharka discovery. PPV-M was the most prevalent strain in peach and apricot. Phylogenetic analyses on (Cter)NIb-(Nter)CP of the isolates were performed. PPV-Rec isolates formed a homogeneous group, while PPV-M isolates split into PPV-Ma and PPV-Mb subgroups. Five separated clades were formed by the analyzed PPV-D isolates. Nucleotide sequences of the partial CP coding region of the analyzed isolates revealed a slightly higher intra-strain genetic variability in PPV-Rec and PPV-M isolates, while that of PPV-D strain isolates was higher from the reported for these strains.