• Title/Summary/Keyword: Genotyping

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

  • Jeon, Chung-Hyun;Park, Sung-Woo
    • Analytical Science and Technology
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    • v.24 no.1
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    • pp.51-59
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    • 2011
  • Genetic Identification has become an important forensic investigation method which discerns identity through analysis of physical samples discovered in various crime scenes. Recently more samples are being requested to undergo A-STR analysis of low copy number (LCN) DNA, which is known as touch evidence-type sample and left on various objects such as a pen briefly used by the criminal, the gear of the car used for driving, the handle, and various buttons inside a car. This research attempted to extract the LCN DNA of the touch evidencetype left on crushed fingerprints on firearms, etc. and examine the genotyping success rate. Four types of firearms (M16, K1A, COLT 45 Pistol, M29 Revolver) were fired individually and physical samples were gathered from four parts of each firearm. Subsequently, in order to extract the LCN DNA, Microkit and $Prepfiler^{TM}$ were used to compare and analyze the quantity of DNA extracted and the genotyping success rate. Analysis results showed that the quantity of DNA extracted by $Prepfiler^{TM}$ was on average 1.7 times higher than that of Microkit, and in genotype analysis success rate $Prepfiler^{TM}$ also demonstrated 24.9% on average in contrast to 0% for Microkit. In regards to the grip part of the K1A, $Prepfiler^{TM}$'s success rate was as high as 50.6%.

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

  • Kim, Hak-Jae;Hahn, Tae-Wook;Juong, Ji-Hun;Bahk, Gyung-Jin;Hong, Chong-Hae
    • Food Science of Animal Resources
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    • v.29 no.6
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    • pp.695-701
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    • 2009
  • This study was performed to evaluate the effectiveness of biological critical control points using the genetic profile of Escherichia coli isolates from pork cutting plants. Samples were collected from carcasses, equipment (knife, table, glove, transport belt, boning and skinning machine), the environment (wall and floor), and meat cuts during the cutting process from two plants. Pulsed-field gel electrophoresis (PFGE) was used to characterize the E. coli isolates. An identical genotype was detected from the carcasses, equipment, environment, and final meat cuts, and showed that the incoming carcasses, which were contaminated during transportation from slaughterhouses, were a major source of E. coli that was spread throughout processing. Also, consistent cross-contamination due to improper cleaning and disinfection procedures was another possibility. As a result, incoming carcasses and cleaning procedures should be considered critical control points in pork cutting plants, since a heating step is not used to inactivate microorganisms. Furthermore, the high rate (59.6%) of E. coli isolation indicates E. coli can be a good indicator in livestock processing plants even though it has genetic diversity.

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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    • v.22 no.5
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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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    • v.37 no.4
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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
    • Horticultural Science & Technology
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    • v.35 no.4
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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.