• 제목/요약/키워드: Genotyping

검색결과 724건 처리시간 0.028초

총기 흔적흔에서의 low copy number(LCN) DNA 검출에 관한 연구 (Research on the detection of LCN DNA from traces on firearms)

  • 전충현;박성우
    • 분석과학
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    • 제24권1호
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    • pp.51-59
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    • 2011
  • 유전자 감식은 다양한 범죄현장에서 발견되는 생체시료의 분석을 통해 신원을 식별하는 중요한 법과학적 수사과정으로 자리 잡았다. 최근에는 범인이 사용했던 펜, 뺑소니 차량에서의 핸들, 기어, 각종 버튼스위치 등에 남겨져 있는 touch evidence-type sample로 알려져 있는 low copy number (LCN) DNA에서의 A-STR분석을 위해 의뢰되는 감정물들이 증가하는 추세에 있다. 본 연구에서는 총기의 뭉개진 지문 등에 남겨져 있는 touch evidence-type의 LCN DNA를 추출하고 유전자형의 분석 성공률을 확인하고 자 하였다. 4종류의 총기(M16, K1A, COLT 45 권총, M29 리볼버)를 각각 격발한 후 총기별로 4곳의 부위에서 시료를 채취한 다음 LCN DNA의 추출을 위해 Microkit과 $Prepfiler^{TM}$ 등 2종류의 시약을 이용하여 DNA 검출량과 유전자형 분석 성공률을 비교 분석하였다. 분석결과 $Prepfiler^{TM}$가 Microkit에 비해 평균 1.7배 DNA검출량이 많았으며, 유전자형 분석 성공률에 있어서도 Microkit은 0%인데 비해 $Prepfiler^{TM}$에서는 평균 24.9%의 성공률을 보였으며, K1A의 손잡이 부위에서 50.6%의 성공률을 나타냈다.

Escherichia coli Genotype을 이용한 생물학적 Critical Control Point의 적합성 평가 (Evaluation of Biological Critical Control Points Using Escherichia coli Genotyping)

  • 김학재;한태욱;정지헌;박경진;홍종해
    • 한국축산식품학회지
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    • 제29권6호
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    • pp.695-701
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    • 2009
  • 본 연구는 축산물가공장 작업환경에 설정된 CCP의 유효성을 평가하기 위하여, 원료도체에서부터 포장에 이르는 전 공정에서 Escherichia coli를 검출하고 PFGE genotyping으로 genetic strain의 흐름을 분석하여 CCP의 적합성을 평가하였다. 시료채취를 위한 작업장은 도축장에서 냉장차량으로 원료도체를 공급받는 독립형 소규모 HACCP 가공장(100두 처리/일)과 축산물종합처리장내 HACCP 가공장(500두 처리/일)을 각각 1곳씩 선정하였다. 시료는 원료도체, 시설 및 장비(발골기, 박피기, 작업대, 벨트, 칼, 면장갑), 환경(벽, 바닥)과 작업공정상의 발골육, 부분육, 최종육을 무균적으로 채취하였다. 작업공정에서 E. coli의 높은 분리율(59.6%)과 작업환경 전반에서의 고른 검출로 E. coli가 축산물가공장 위생지표균으로의 활용 가능성을 확인할 수 있었다. 분리된 E. coli의 genotype 분포는 작업장내 미생물 오염의 흐름을 보여주었다. 소규모가공장의 경우 오염된 원료육 반입이 작업공정을 오염시키고 공정중의 교차오염 발생으로 최종육에 영향을 주었고, 축산물종합처리장내 작업장은 원료육 오염보다는 작업환경에 상존하는 오염미생물의 교차오염이 최종육에 영향을 주는 양상이었다. 따라서 포장육가공장과 같이 작업공정에 멸균처리 과정이 없는 경우는 선행요건에 해당되는 원료반입과 작업환경의 세척 및 소독이 CCP로 관리되거나 이에 준하는 특별관리가 필요하였다. 결과적으로, 작업공정에서의 E. coli 검출과 genotyping은 작업공정상의 미생물 오염의 흐름을 파악하는데 유용하였으며, 이러한 방법은 HACCP 작업장의 미생물 오염원 분석과 설정된 생물학적 critical control point의 적합성 평가에 효과적이었다.

Immunosuppression-enhancing effect of the administration of allogeneic canine adipose-derived mesenchymal stem cells (cA-MSCs) compared with autologous cA-MSCs in vitro

  • Wi, Hayeon;Lee, Seunghoon;Kim, Youngim;No, Jin-Gu;Lee, Poongyeon;Lee, Bo Ram;Oh, Keon Bong;Hur, Tai-young;Ock, Sun A
    • Journal of Veterinary Science
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    • 제22권5호
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    • pp.63.1-63.14
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    • 2021
  • Background: Recently, mesenchymal stem cells therapy has been performed in dogs, although the outcome is not always favorable. Objectives: To investigate the therapeutic efficacy of mesenchymal stem cells (MSCs) using dog leukocyte antigen (DLA) matching between the donor and recipient in vitro. Methods: Canine adipose-derived MSCs (cA-MSCs) isolated from the subcutaneous tissue of Dog 1 underwent characterization. For major DLA genotyping (DQA1, DQB1, and DRB1), peripheral blood mononuclear cells (PBMCs) from two dogs (Dogs 1 and 2) were analyzed by direct sequencing of polymerase chain reaction (PCR) products. The cA-MSCs were co-cultured at a 1:10 ratio with activated PBMCs (DLA matching or mismatching) for 3 days and analyzed for immunosuppressive (IDO, PTGS2, and PTGES), inflammatory (IL6 and IL10), and apoptotic genes (CASP8, BAX, TP53, and BCL2) by quantitative real-time reverse transcriptase-PCR. Results: cA-MSCs were expressed cell surface markers such as CD90+/44+/29+/45- and differentiated into osteocytes, chondrocytes, and adipocytes in vitro. According to the Immuno Polymorphism Database, DLA genotyping comparisons of Dogs 1 and 2 revealed complete differences in genes DQA1, DQB1, and DRB1. In the co-culturing of cA-MSCs and PBMCs, DLA mismatch between the two cell types induced a significant increase in the expression of immunosuppressive (IDO/PTGS2) and apoptotic (CASP8/BAX) genes. Conclusions: The administration of cA-MSCs matching the recipient DLA type can alleviate the need to regulate excessive immunosuppressive responses associated with genes, such as IDO and PTGES. Furthermore, easy and reliable DLA genotyping technology is required because of the high degree of genetic polymorphisms of DQA1, DQB1, and DRB1 and the low readability of DLA 88.

Prevention of thiopurine-induced early leukopenia in a Korean pediatric patient with Crohn's disease who turned out to possess homozygous mutations in NUDT15 R139C

  • Bae, Jaewoan;Choe, Byung-Ho;Kang, Ben
    • Journal of Yeungnam Medical Science
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    • 제37권4호
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    • pp.332-336
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    • 2020
  • Homozygous mutations in NUDT15 R139C are known as the major factor associated with thiopurine-induced early leukopenia, particularly in Asian patients. Therefore, NUDT15 genotyping is currently recommended before thiopurine treatment to identify patients who are NUDT15 poor metabolizers and consider the use of an alternative immunomodulatory therapy. We report a case of a 12-year-old Korean girl with Crohn's disease (CD), in whom thiopurine-induced leukopenia was prevented by initiation of azathioprine (AZA) therapy at a low dose (0.5 mg/kg/day) and early detection of significant hair loss and white blood cell (WBC) count decrease at 17 days from the start of AZA treatment. The WBC count dropped from 8,970/μL to 3,370/μL in 2 weeks, and AZA treatment was stopped because of concerns of potential leukopenia in the near future. Her WBC count recovered to 5,120/μL after 3 weeks. Gene analysis later revealed that she had a homozygous mutation in NUDT15 R139C, resulting in a poor metabolizing activity of NUDT15. In situations when NUDT15 genotyping is unavailable, initiation of AZA therapy at 0.5 mg/kg/day with close observation of hair loss and WBC counts within 2 weeks may be an alternative way to prevent thiopurine-induced early leukopenia in Asian children with CD.

Development of Fluidigm SNP Type Genotyping Assays for Marker-assisted Breeding of Chili Pepper (Capsicum annuum L.)

  • Kim, Haein;Yoon, Jae Bok;Lee, Jundae
    • 원예과학기술지
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    • 제35권4호
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    • pp.465-479
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    • 2017
  • Chili pepper (Capsicum annuum L.) is an economically important horticultural crop in Korea; however, various diseases, including Phytophthora root rot, anthracnose, powdery mildew, Cucumber mosaic virus (CMV), Pepper mild mottle virus (PMMoV), and Pepper mottle virus (PepMoV), severely affect their productivity and quality. Therefore, pepper varieties with resistance to multiple diseases are highly desired. In this study, we developed 20 SNP type assays for three pepper populations using Fluidigm nanofluidic dynamic arrays. A total of 4,608 data points can be produced with a 192.24 dynamic array consisting of 192 samples and 24 SNP markers. The assays were converted from previously developed sequence-tagged-site (STS) markers and included markers for resistance to Phytophthora root rot (M3-2 and M3-3), anthracnose (CcR9, CA09g12180, CA09g19170, CA12g17210, and CA12g19240), powdery mildew (Ltr4.1-40344, Ltr4.2-56301, and Ltr4.2-585119), bacterial spot (Bs2), CMV (Cmr1-2), PMMoV (L4), and PepMoV (pvr1 and pvr2-123457), as well as for capsaicinoids content (qcap3.1-40134, qcap6.1-299931, qcap6.1-589160, qdhc2.1-1335057, and qdhc2.2-43829). In addition, 11 assays were validated through a comparison with the corresponding data of the STS markers. Furthermore, we successfully applied the assays to commercial $F_1$ cultivars and to our breeding lines. These 20 SNP type assays will be very useful for developing new superior pepper varieties with resistance to multiple diseases and a higher content of capsaicinoids for increased pungency.