• 제목/요약/키워드: genotyping techniques

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유전체 코호트 연구를 위한 대용량 염기서열 분석 (High Throughput Genotyping for Genomic Cohort Study)

  • 박웅양
    • Journal of Preventive Medicine and Public Health
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    • 제40권2호
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    • pp.102-107
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    • 2007
  • Human Genome Project (HGP) could unveil the secrets of human being by a long script of genetic codes, which enabled us to get access to mine the cause of diseases more efficiently. Two wheels for HGP, bioinformatics and high throughput technology are essential techniques for the genomic medicine. While microarray platforms are still evolving, we can screen more than 500,000 genotypes at once. Even we can sequence the whole genome of an organism within a day. Because the future medicne will focus on the genetic susceptibility of individuals, we need to find genetic variations of each person by efficient genotyping methods.

Molecular typing of uropathogenic Escherichia coli isolated from Korean children with urinary tract infection

  • Yun, Ki Wook;Kim, Do Soo;Kim, Wonyong;Lim, In Seok
    • Clinical and Experimental Pediatrics
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    • 제58권1호
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    • pp.20-27
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    • 2015
  • Purpose: We investigated the molecular types of uropathogenic Escherichia coli (UPEC) by using conventional phylogrouping, multilocus sequence typing (MLST), and fimH genotyping. Methods: Samples of patients younger than 18 years of age were collected from the Chung-Ang University Hospital over 2 years. Conventional phylogenetic grouping for UPEC strains was performed by polymerase chain reaction (PCR). Bacterial strain sequence types (STs) were classified on the basis of the results of partial sequencing of seven housekeeping genes. In addition, we analyzed nucleotide variations in a 424-base pair fragment of fimH, a major virulence factor in UPEC. Results: Sixty-four UPEC isolates were analyzed in this study. Phylogenetic grouping revealed that group B2 was the most common type (n=54, 84%). We identified 16 distinctive STs using MLST. The most common STs were ST95 (35.9%), ST73 (15.6%), ST131 (12.5%), ST69 (7.8%), and ST14 (6.3%). Fourteen fimH allele types were identified, of which 11 had been previously reported, and the remaining three were identified in this study. f1 (n=28, 45.2%) was found to be the most common allele type, followed by f6 and f9 (n=7, 11.3% each). Comparative analysis of the results from the three different molecular typing techniques revealed that both MLST and fimH typing generated more discriminatory UPEC types than did PCR-based phylogrouping. Conclusion: We characterized UPEC molecular types isolated from Korean children by MLST and fimH genotyping. fimH genotyping might serve as a useful molecular test for large epidemiologic studies of UPEC isolates.

Clinical Application of ABO Genotyping: 10 Years' Experience in the Southeastern Korea

  • Sae Am Song;Eun-Kyung Yu;Seung Hwan Oh
    • Journal of Interdisciplinary Genomics
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    • 제6권1호
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    • pp.6-13
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    • 2024
  • Background: ABO typing is crucial for ensuring safe blood transfusion and is commonly performed by examining antigen-antibody interactions. Determining ABO blood group can be difficult when dealing with ABO discrepancy and ABO subgroups. ABO genotyping may be necessary to resolve ABO discrepancy. ABO genotyping primarily involves direct sequencing, with the possibility of using other molecular methods. Methods: PCR and direct sequencing of exons 6 and 7 were performed for total 108 samples from June 2010 to December 2019. Also, other molecular methods including cloning sequencing and short tandem repeat analysis were carried out just in case. Sequencing data were compared with allele information of blood group antigen mutation databases. Results: The predominant causal allele among 108 ABO discrepant cases was cis-AB01, with 28 cases. This was followed by rare ABO alleles (B309, B306, A204, Bw29, and Ax01) with 14 cases, and blood chimera with 5 cases. Five new alleles were identified during the investigation. Conclusion: This study reaffirms that cis-AB is the most common cause of inherited ABO discrepancies, and cis-AB01 is the most prevalent cis-AB allele in the Korean population, also in the southeastern region. In addition, we discovered five new alleles and five blood chimeras by adopting sequencing analysis and additional molecular techniques to resolve ABO discrepancies, which provide regional data on rare alleles. This study presents rare and new ABO alleles and blood chimeras identified over a ten-year period at two major university hospitals in Southeastern Korea.

Determining genetic diversity of prevalent G and P genotype of Bovine Rotavirus A from neonatal calves of Gujarat, India

  • Akash Golaviya;Rafiyuddin Mathakiya;Subhash Jakhesara;Prakash Koringa
    • Journal of Veterinary Science
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    • 제25권4호
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    • pp.55.1-55.12
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    • 2024
  • Importance: Neonatal calf diarrhea is a major cause of mortality in newborn calves worldwide, posing a significant challenge in bovine herds. Group A Bovine Rotaviruses (BRVA) are the primary contributors to severe gastroenteritis in calves under two months old. Objectives: This study examined the prevalence and molecular characterization of BRVA in neonatal calves in Gujarat, India. Methods: Sixty-nine diarrheic fecal samples were collected and subjected to various molecular methods of BRVA detection, isolation, and characterization. Results: The latex agglutination test (LAT), electropherotyping (RNA-PAGE), and reverse transcription polymerase chain reaction revealed positivity rates of 39.13%, 20.30%, and 37.70%, respectively. RNA-PAGE identified 11 bands with a 4:2:3:2 migration pattern, indicative of the segmented genome of BRVA. BRVA was successfully isolated from LATpositive samples, with 26 samples exhibiting clear cytopathic effects upon passage in MA-104 cell lines. Genotyping identified G10 as the predominant G genotype, with P[11] genotypes comprising 76.92% of the isolates. The most common G/P combination was G10P[11], highlighting its zoonotic potential. Conclusions and Relevance: These findings underscore the importance of molecular detection and genotyping for effective vaccine development. This study provides crucial insights into the prevalent G and P genotypes of BRVA in Gujarat, India, aiding in the development of targeted control measures.

Digital image-based plant phenotyping: a review

  • Omari, Mohammad Kamran;Lee, Jayoung;Faqeerzada, Mohammad Akbar;Joshi, Rahul;Park, Eunsoo;Cho, Byoung-Kwan
    • 농업과학연구
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    • 제47권1호
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    • pp.119-130
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    • 2020
  • With the current rapid growth and increase in the world's population, the demand for nutritious food and fibers and fuel will increase. Therefore, there is a serious need for the use of breeding programs with the full potential to produce high-yielding crops. However, existing breeding techniques are unable to meet the demand criteria even though genotyping techniques have significantly progressed with the discovery of molecular markers and next-generation sequencing tools, and conventional phenotyping techniques lag behind. Well-organized high-throughput plant phenotyping platforms have been established recently and developed in different parts of the world to address this problem. These platforms use several imaging techniques and technologies to acquire data for quantitative studies related to plant growth, yield, and adaptation to various types of abiotic or biotic stresses (drought, nutrient, disease, salinity, etc.). Phenotyping has become an impediment in genomics studies of plant breeding. In recent years, phenomics, an emerging domain that entails characterizing the full set of phenotypes in a given species, has appeared as a novel approach to enhance genomics data in breeding programs. Imaging techniques are of substantial importance in phenomics. In this study, the importance of current imaging technologies and their applications in plant phenotyping are reviewed, and their advantages and limitations in phenomics are highlighted.

식물유전 및 육종학 연구에서의 분자생물학적 마커기술의 이용 (Utilization of Molecular Markers in Plant Genetics and Breeding)

  • 이주경
    • 한국자원식물학회지
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    • 제10권2호
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    • pp.200-210
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    • 1997
  • The understanding on the plant genome is accelerated with the fast advance of molecular biological techniques. The molecular dissecting of the plant genome has made possible the precise genotyping the plants, which can be utilized for molecular breeding program. As well, the molecular cloning of genes interested can facilitate the process of gene transfer between intra-and inter-generic taxa. Moreover, the manipulation of the agronomically important QTL genes, which can be rarely performed by the conventional genetic methods, is also possible by the utilization of molecular markers. In addition to these genetical applications, molecular markers are useful in the areas of plant taxonomy and management of germplasm by fingerprinting analysis. This paper describes the theoretical aspects marker technologies and practical applications of each marker technique.

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Novel High-Throughput DNA Part Characterization Technique for Synthetic Biology

  • Bak, Seong-Kun;Seong, Wonjae;Rha, Eugene;Lee, Hyewon;Kim, Seong Keun;Kwon, Kil Koang;Kim, Haseong;Lee, Seung-Goo
    • Journal of Microbiology and Biotechnology
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    • 제32권8호
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    • pp.1026-1033
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    • 2022
  • This study presents a novel DNA part characterization technique that increases throughput by combinatorial DNA part assembly, solid plate-based quantitative fluorescence assay for phenotyping, and barcode tagging-based long-read sequencing for genotyping. We confirmed that the fluorescence intensities of colonies on plates were comparable to fluorescence at the single-cell level from a high-end, flow-cytometry device and developed a high-throughput image analysis pipeline. The barcode tagging-based long-read sequencing technique enabled rapid identification of all DNA parts and their combinations with a single sequencing experiment. Using our techniques, forty-four DNA parts (21 promoters and 23 RBSs) were successfully characterized in 72 h without any automated equipment. We anticipate that this high-throughput and easy-to-use part characterization technique will contribute to increasing part diversity and be useful for building genetic circuits and metabolic pathways in synthetic biology.

Comparison of Different PCR-Based Genotyping Techniques for MRSA Discrimination Among Methicillin-Resistant Staphylococcus aureus Isolates

  • Kim, Keun-Sung;Seo, Hyun-Ah;Oh, Chang-Yong;Kim, Hong
    • Journal of Microbiology and Biotechnology
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    • 제11권5호
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    • pp.788-797
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    • 2001
  • The usefulness of three PCR methods were evaluated for the epidemiological typing of Staphylococcus aureus: an enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR), repetitive extragenic palindromic element PCR (REP-PCR), and 16S-23S intergenic spacer PCR (ITS-PCR). The analysis was performed using a collection of S. aureus strains comprised of 6 reference and 79 isolates from patients with various diseases. Among the 85 S. aureus strains tested, 6 references and 6 isolates were found to be susceptible to methicillin, whereas the remaining 73 isolates were resistant to it. PCR methods are of special concern, as conventional phenotypic methods are unable to clearly distinguish among methicillin-resistant S. aureus (MRSA) strains. The ability of the techniques to detect different unrelated types was found to be as follows: ERIC-PCR, 19 types; REP-PCR, 36 types; and ITS-PCR, 14 types. On the basis of combining the ERIC, REP, and ITS fingerprints, the 85 S. aureus strains were grouped into 56 genetic types (designated G1 to G56). The diversities for the 85 S. aureus strains, calculated according to Simpson\`s index, were 0.88 for an ERIC-PCR, 0.93 for a REP-PCR, and 0.48 for an ITS-PCR, and the diversity increased up to 0.97 when an ERIC-PCR and REP-PCR were combined. The above discrimination indices imply that the genetic heterogeneity of S. aureus strains is high. Accordingly, this study demonstrates that DNA sequences from highly conserved repeats of a genome, particularly a combination of ERIC sequences and REP elements, are a convenient and accurate tool for the subspecies-specific discrimination and epidemiologic tracking of S. aureus.

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