• Title/Summary/Keyword: DNA typing

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Random amplification of polymorphic DNA typing of Listeria monocytogenes isolates from animal products (축산물유래 Listeria monocytogenes의 RAPD typing)

  • Lee, Chul-Hyun;Son, Won-Geun
    • Korean Journal of Veterinary Research
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    • v.49 no.4
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    • pp.319-328
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    • 2009
  • This study investigated the epidemiology of Listeria (L.) monocytogenes, a food-borne pathogen. The epidemiology of food-borne pathogens is of great importance for clarifying bacterial origin and preventing bacterial contamination and infection. This work examined 68 L. monocytogenes strains, including 11 reference strains and 57 isolates from imported US beef, domestic meats (beef, pork, chicken meat), raw milk, and milk plants. The random amplified polymorphic DNA (RAPD) techniques were optimized to develop a standard molecular epidemiological analysis of L. monocytogenes. There was great genetic variability among the isolates, which produced 24 and 34 RAPD patterns with primer HLWL85 and HLWL74, respectively. The discriminatory power of the RAPD methods with HLWL85 and HLWL74 primer were very high (DI = 0.957; S ${\geq}$ 80%, S ${\geq}$ 95%). Some RAPD types were specific to origin. A few RAPD types were specific for L. monocytogenes strains belonging to a particular serotype. Using the HLWL85 primer, the strains isolated from milk plants could be distinguished from the other strains. And using the HLWL74 primer, the strains isolated from imported beef (US) could be distinguished completely from the other strains.

A Case of Parentage Testing in Dog by Microsatellite DNA Typing (Microsatellite DNA형에 의한 개의 친자감정예)

  • 조길재;조병욱;이길왕;김선구;김용균
    • Journal of Life Science
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    • v.13 no.4
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    • pp.416-420
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    • 2003
  • We performed this study to determine the parentage verification of putative dogs. A total of 7 samples (4 Labrador Retriever dog and 3 Poongsan dog) were genotyped by using 12 international markers (PEZ1, PEZ5, PEZ8, PEZ10, PEZ11, PEZ12, PEZ17, PEZ20, PEZ21, FHC2010, FHC2054, FHC2079). This methods consisted of multiplexing PCR procedures, and it showed reasonable amplification of all PCR products. Genotyping was performed with an ABI 310 genetic analyzer. Labrador Retriever Pup I and Pup II were included according to principles of Mendelian genetics in all loci, while Poongsan Pup III was excluded with markers PEZ1 (106/118), PEZ10 (276/300), and FHC2010 (228/232). These results suggest that the present DNA typing is so useful for parentage verification of these two breeds.

Microbial Forensics: Human Identification

  • Eom, Yong-Bin
    • Biomedical Science Letters
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    • v.24 no.4
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    • pp.292-304
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    • 2018
  • Microbes is becoming increasingly forensic possibility as a consequence of advances in massive parallel sequencing (MPS) and bioinformatics. Human DNA typing is the best identifier, but it is not always possible to extract a full DNA profile namely its degradation and low copy number, and it may have limitations for identical twins. To overcome these unsatisfactory limitations, forensic potential for bacteria found in evidence could be used to differentiate individuals. Prokaryotic cells have a cell wall that better protects the bacterial nucleoid compared to the cell membrane of eukaryotic cells. Humans have an extremely diverse microbiome that may prove useful in determining human identity and may even be possible to link the microbes to the person responsible for them. Microbial composition within the human microbiome varies across individuals. Therefore, MPS of human microbiome could be used to identify biological samples from the different individuals, specifically for twins and other cases where standard DNA typing doses not provide satisfactory results due to degradation of human DNA. Microbial forensics is a new discipline combining forensic science and microbiology, which can not to replace current STR analysis methods used for human identification but to be complementary. Among the fields of microbial forensics, this paper will briefly describe information on the current status of microbiome research such as metagenomic code, salivary microbiome, pubic hair microbiome, microbes as indicators of body fluids, soils microbes as forensic indicator, and review microbial forensics as the feasibility of microbiome-based human identification.

Molecular Typing of Acinetobacter Baumannii Strains by Randomly Amplified Polymorphic DNA (RAPD) Analysis (Randomly Amplified Polymorphic DNA (RAPD) 분석에 의한 Acinetobacter Baumannii 균주의 유전형 분류)

  • Oh, Jae-Young;Cho, Jae-Wee;Park, Jong-Chun;Lee, Je-Chul
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.2
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    • pp.129-139
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    • 2000
  • Acinetobacter baumannii strains are emerging pathogens of the nosocomial infection with an increasing frequency in recent years. The therapeutic difficulty due to the wide spread of multiple resistant strains was major problem in A. baumannii infection. It seems likely that high frequency of A. baumannii infection will be increasing epidemiological importance in the future. However, the current limited understanding of the epidemiology of A. baumannii infections is caused by lack of a rapid and practical method for the molecular characterization of A. baumannii strains. This study was undertaken to determine molecular types and genetic similarity among A. baumannii strains isolated from four hospitals by RAPD analysis. Eighty-five strains, including 40 from Chunnam University Hospital, 27 from Dankook University Hospital, 15 from Yonsei University Hospital, and 3 from Seonam University Hospital, were classified into three molecular types. Molecular type II was the most common pattern and included 72 strains. All strains from Dankook University Hospital and 40 strains from Chunnam University Hospital belonged to molecular type I or II. A. baumannii strains form Yonsei University Hospital were very distant similarity values. The range of genetic similarity values among 85 strains of A. baumannii was 0.26 to 1.00. Although phenotypes including biotype and antimicrobial resistance pattern of A. baumannii strains were same or very similar to each other, their RAPD patterns were quite different. Typing with phenotypes was found to be less reliable than molecular typing by RAPD analysis. These results suggest that RAPD analysis provides rapid and simple typing method of A. baumannii strains for epidemiological studies. This work is the first epidemiological report of A. baumannii infections in Korea and it is hoped that results of this work may contribute to a better understanding of the clinical importance and epidemiology of A. baumannii strains.

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Microsatellite DNA Typing Using 16 Markers for Parentage Verification of the Korean Native Horse

  • Cho, G.J.;Cho, B.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.6
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    • pp.750-754
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    • 2004
  • This study was performed for parentage verification of the Korean native horse (KNH). 103 random KNH samples (including 19 foals for parentage testing) were genotyped by using 16 microsatellite markers. The number of alleles per locus varied from 5 to 13 with an average value of 8.56 in the KNH. The observed heterozygosity and the expected heterozygosity ranged 0.398-0.893 (the average value was 0.683) and 0.368-0.871 (the average value was 0.727) in the KNH, respectively. The PIC value and the exclusion probability ranged 0.347-0.853 (the average value was 0.692) and 0.208-0.736, respectively, and the total exclusion probability of 16 microsatellite loci was 0.9999. Of the 16 markers, AHT4, AHT5, ASB2, ASB17, HMS2, HMS3, HTG10, LEX33, TKY321 and VHL20 loci have a relatively high PIC value (>0.7) in the KNH. Of the 19 foals, 5 foals were disqualified by an incompatibility of 4-7 markers according to a Mendelian fashion in the present DNA typing for parentage testing. These results present basic information for developing a system for parentage verification and individual identification in the KNH.

Random amplified polymorphic DNA analysis of bacterial pathogens using universal rice primers

  • Monoldorova, Sezim;Kim, Jinsol;Kim, Joon Hee;Jeon, Bo-Young
    • Korean Journal of Veterinary Service
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    • v.40 no.1
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    • pp.1-6
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    • 2017
  • Molecular typing of pathogenic microorganisms is important for epidemiological investigation of infectious disease outbreaks. In this study, we applied Universal Rice Primers (URP) that were originated from repetitive sequences in rice chromosomal DNA to random amplified polymorphic DNA (RAPD) analysis of pathogenic bacteria such as Escherichia coli, Listeria monocytogenes, and Salmonella sp. Of the twelve URP primers examined to date, seven primers (URP-2, -3, -4, -5, -6, -8, and -9) generated reproducible and polymorphic PCR products ranging from 1 to 13 bands. One of them, URP-6 was very effective in differentiating seven E. coli serotypes, seven L. monocytogenes clinical isolates, and eight Salmonella subspecies (ssp.) serovars. The results thus indicate that RAPD analysis using URP primers might be useful in typing bacterial pathogens including E. coli, L. monocytogenes, and Salmonella strains.

Random Amplified Polymorphic DNA-PCR Analysis for Identification of Bacillus anthracis (탄저균의 Random Amplified Polymorphic DNA-PCR 분석)

  • 김성주;박경현;김형태;조기승;김기천;최영길;박승환;이남택;채영규
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.56-60
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    • 2001
  • Molecular typing of Bacillus anthracis has been extremely difficult due to the lack of polymorphic DNA markers. Aiming to develop a DNA marker specific for Bacillus anthracis and to be able to discriminate this species from Bacillus genus, we applied the random amplified polymorphic DNA (RAPD)-PCR. We have identified B. anthracis from various Bacillus species. The analysis performed by RAPD clearly demonstrated substantial genetic variations among Bacillus species. This type of analysis is an easy, quick and highly discriminatory technique that may help in diagnosis of anthrax.

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Genetic Characteristics of mtDNA and STR marker in Human Bone Excavated from Mokgam-dong, Siheung in Korea (시흥 목감동 출토 인골의 미토콘드리아 DNA와 STR의 유전적 특징)

  • Seo, Min-Seok;Chung, Yong-Jae;Lee, Kyu-Shik;Park, Ki-Won
    • 보존과학연구
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    • s.24
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    • pp.153-167
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    • 2003
  • We performed nuclear DNA typing and mitochondrial DNA sequencing analysis based on PCR from an ancient Korean remainsexcavated from Siheung in Korea. 7 bones were collected and partially STR(short tandem repeat) systems, Sex determination Amelogenin kit(Promega co, USA), were used in this study. Mitochondrial DNAs were also amplified and sequenced by ABI 310 DNA sequencer. We know that sample no. 2 and no. 3 were females and also sample no. 2 and no.7 possessed the same maternal inheritance by mitochondrial DNA sequencing results. Throughout this research, the mitochondrial DNA sequencing of human in the middle of Joseon Dynasty in Korea is obtained. In addition, this finding will be an important foundation for the future research.

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Evaluation of DNA Extraction Methods from Low Copy Number (LCN) DNA Samples for Forensic DNA Typing

  • Eom, Yong-Bin
    • Biomedical Science Letters
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    • v.15 no.3
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    • pp.229-232
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    • 2009
  • DNA isolation for PCR-based short tandem repeat (STR) analysis is essential to recover high yields of amplifiable DNA from low copy number (LCN) DNA samples. There are different methods developed for DNA extraction from the small bloodstain and gloves, commonly found at crime scenes. In order to obtain STR profiles from LCN DNA samples, DNA extraction protocols, namely the automated $iPrep^{TM}$ $ChargeSwitch^{(R)}$ method, the automated $QIAcube^{TM}$ method, the automated $Maxwell^{(R)}$ 16 DNA $IQ^{TM}$ Resin method, and the manual $QIAamp^{(R)}$ DNA Micro Kit method, were evaluated. Extracted DNA was quantified by the $Quantifiler^{TM}$ Human DNA Quantification Kit and DNA profiled by $AmpFISTR^{(R)}$ $Identifiler^{(R)}$ Kit. Results were compared based on the amount of DNA obtained and the completeness of the STR profiles produced. The automated $iPrep^{TM}$ $ChargeSwitch^{(R)}$ and $QIAcube^{TM}$ methoas produced reproducible DNA of sufficient quantity and quality trom the dried blood spot. This two automated methods showed a quantity and quality comparable to those of the forensic manual standard protocols normally used in our laboratory. In our hands, the automated DNA extraction method is another obvious choice when the forensic case sample available is bloodstain. The findings of this study indicate that the manual simple modified $QIAamp^{(R)}$ DNA Micro Kit method is best method to recover high yields of amplifiable DNA from the numerous potential sources of LCN DNA samples.

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