• Title/Summary/Keyword: Species detection

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PCR Based Detection of Phellinus linteus using Specific Primers Generated from Universal Rice Primer(URP) Derived PCR Polymorphic Band

  • Kang, Hee-Wan;Park, Dong-Suk;Park, Young-Jin;Lee, Byoung-Moo;Cho, Soo-Muk;Kim, Ki-Tae;Seo, Geon-Sik;Go, Seung-Joo
    • Mycobiology
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    • v.30 no.4
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    • pp.202-207
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    • 2002
  • This study was carried out to develop specific primers for PCR detection of Phellinus linteus. Diverse genomes of 15 Phellinus spp. including five Phellinus linteus isolates were fingerprinted by Primer Universal rice primer(URP)1F. The URP-PCR pattern differentiated P. linteus isolates from other phellinus spp. A polymorphic band(2.8 kb), which is unique for P. linteus isolates, was isolated and sequenced. Twenty four-oligonucleotide primer pairs were designed based on information of DNA sequence. The primer set(PLSPF2/PLSPR1) amplified single band(2.2 kb) of expected size with genomic DNA from seven Phellinus linteus, but not with that of other Phellinus species tested. The primers could be used identically in both DNA samples from mycelium and fruit bodies. This specific primers could offer a useful tool for detecting and identifying P. linteus rapidly.

First Detection of Penicillium fellutanum from Stored Rice in Korea

  • Oh, Ji-Yeon;Sang, Mee-Kyung;Lee, Ho-Joung;Ryoo, Mun-Il;Kim, Ki-Deok
    • Research in Plant Disease
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    • v.17 no.2
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    • pp.216-221
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    • 2011
  • A representative isolate KU53 of the predominant Penicillium species was obtained from rice samples from rice processing complexes of National Agricultural Cooperative Federation in Korea. In this study, isolate KU53 was identified by its morphological and molecular characteristics. The macro- and microscopic characteristics of isolate KU53 were compared with the P. fellutanum reference isolate KCTC16913 on different media; isolate KU53 was generally identical to those of the reference isolate KCTC16913. In a molecular-based identification, the ${\beta}$-tubulin and translation elongation factor 1-alpha sequences of isolate KU53 was most closely related to those of P. fellutanum. Thus, isolate KU53 from stored rice could be identified as P. fellutanum, some isolates of which are known to produce mycotoxin-related metabolites. To our knowledge, this is the first detection of P. fellutanum from stored rice in Korea.

Detection of Mold by Enzyme-Linked Immunosorbent Assay

  • Kwak, Bo-Yeon;Kim, Soon-Young;Shon, Dong-Hwa
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.764-772
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    • 1999
  • To develop an enzyme-linked immunosorbent assay (ELISA) for detecting mold, we produced anti-mold polyclonal antibodies by immunizing extracellular polysaccharide (EPS) of Aspergillus flavus or Penicillium citrinum in rabbits subcutaneously. Using the purified antibody (Ab) and Ab-HRP conjugate, a sandwich ELISA for EPS was established. The standard curve of the ELISA showed the detection limit for P citrinum EPS to be $0.003{\;}\mu\textrm{g}/ml$. The cross-reactivities of the anti-P citrinum EPS Ab toward components of P citrinum such as EPS, liquid, and solid culture mycelium were 100, 10.5, and 0.58%, respectively, and those toward components of A. flavus such as EPS, liquid and solid culture mycelium, and spore were 300, 0.67, 0.29, and 0%, respectively. When the reactivities toward culture broths of 59 mold strains were tested by the sandwich ELISA, most of the Aspergillus (16 of 18) and Penicillium (14 of 16) strains along with one of the two Cladosporium strains gave positive signals in the culture broths even when diluted 1,000 fold, while the rest of species such as Fusarium, Absidia, Alternaria, and Candida gave negative signals. When the water extracts of 30 corn samples were analyzed by the sandwich ELISA, the EPS in the com could be detected in the concentration range of $0.1-1.6{\;}\mu\textrm{g}/g$.

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Gas-Sensing Membrane Electrodes for the Determination of Dissolved Gases (I). Continuous-Automated Determination of Nitrite Ion Using Tubular PVC Membrane Type of pH Electrode (용해기체 분석용 기체 감응막 이온선택성 전극 (제 1 보). 관형 PVC 막 pH 전극을 이용한 아질산이온의 연속·자동화 정량)

  • Heung Lark Lee;Jong Hoon Yun
    • Journal of the Korean Chemical Society
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    • v.33 no.4
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    • pp.388-398
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    • 1989
  • A continuous-automated method for the determination of nitrite ion using gas-sensing membrane electrode was developed. The pH electrode of tubular PVC membrane type was used as a detector of this system. The slope of linear response of the electrode measured at optimum conditions for the continuous-automated determination of nitrite ion was 63.5 mV/decade. The concentration range of linear response and detection limit were 2.5 ${\times}10^{-4}{\sim}\;7.5{\times}10^{-2}$M and $8.0{\times}10^{-5}$M, respectively. This detection system was not only less interfering to acidic gas species than other methods but also less time consumable for determination.

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Detection of Methicillin Resistance in Staphylococcus aureus Isolates Using Two-Step Triplex PCR and Conventional Methods

  • Cho, Joon-Il;Jung, Hye-Jin;Kim, Young-Joon;Park, Sung-Hee;Ha, Sang-Do;Kim, Keun-Sung
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.673-676
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    • 2007
  • A two-step triplex PCR assay targeting the mecA, femA, and nuc genes was developed for the detection of methicillin resistance genes harbored by some Staphylococcus aureus isolates and for the simultaneous identification of such isolates at the species level. The triplex PCR revealed the presence of the femA and nuc genes in all the S. aureus isolates examined (n=105). Forty-four clinical isolates were mecA positive and no foodborne isolates were mecA positive. The PCR results had a 98 or 99% correlation with the results of PBP2a latex agglutination tests or oxacillin susceptibility tests, respectively.

Simple Detection of Cochliobolus Fungal Pathogens in Maize

  • Kang, In Jeong;Shim, Hyeong Kwon;Roh, Jae Hwan;Heu, Sunggi;Shin, Dong Bum
    • The Plant Pathology Journal
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    • v.34 no.4
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    • pp.327-334
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    • 2018
  • Northern corn leaf spot and southern corn leaf blight caused by Cochliobolus carbonum (anamorph, Bipolaris zeicola) and Cochliobolus heterostrophus (anamorph, Bipolaris maydis), respectively, are common maize diseases in Korea. Accurate detection of plant pathogens is necessary for effective disease management. Based on the polyketide synthase gene (PKS) of Cochliobolus carbonum and the nonribosomal peptide synthetase gene (NRPS) of Cochliobolus heterostrophus, primer pairs were designed for PCR to simultaneously detect the two fungal pathogens and were specific and sensitive enough to be used for duplex PCR analysis. This duplex PCR-based method was found to be effective for diagnosing simultaneous infections from the two Cochliobolus species that display similar morphological and mycological characteristics. With this method, it is possible to prevent infections in maize by detecting infected seeds or maize and discarding them. Besides saving time and effort, early diagnosis can help to prevent infections, establish comprehensive management systems, and secure healthy seeds.

LASER FLASH PHOTOLYSIS STUDY ON THE PHOTOCYCLIZATION OF N-(O-HALOBENZYL) IMIDAZOLE

  • Park, Yong-Tae;Hwang, Young-Sun;Song, Woong Song;Kim, Dongho
    • Journal of Photoscience
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    • v.3 no.2
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    • pp.91-93
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    • 1996
  • In connection with our interest on the photochemical properties of heteroaryl halides, which are currently the subject of heterocyclic ring formation and haloarene degradation, we have studied the photochemistry of the haloarene linked to N-heteroarene compounds. Imidazo[5,1-a]isoindole was synthesized from N-(ochlorobenzyl)imidazole or N-(o-bromobenzyl) imidazole in acidic aqueous solution or acetonitrile via the intramolgcular photocyclization (Table 1). This type of reaction provides the synthetic methods for 5- and 6-membered polyheteroatomic heterocyclic ring compounds. However, the reaction mechanism for the intramolecular photocyclization of haloarene tethered heteroarenes has not yet been established. Grimshaw et al. suggested a mechanism for homolyric carbonhalogen bond fission assisted by radical complexation to explain their results in the photocyclization of 5-(2-chlorophenyl)-1,3-diphenylpyrazole. They also reported the detection of acyclohexadienyl intermediate involved in the above reaction. Park et al. reported several transient 'intermediates involved in the laser flash photolysis of N-(o-halobenzyl) pyridinium and N-benzyl-2-halopyridinium salts. Thus we performed the laser flash photolysis study on the photocyclization reaction of N-(o-chlorobenzyl) imidazole to identify the intermediate species involved in the reaction. Here, we report on the preliminary results in the photocyclization reaction of N-(o-halobenzyl)imidazole through the detection of reaction intermediates.

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Detection of Colletotrichum acutatum and C. gloeosporioides by Real Time PCR (Real Time PCR을 이용한 Colletotrichum acutatum과 C. gloeosporioides의 검출)

  • Kim, Seung-Han;Kwon, Oh-Hun
    • Research in Plant Disease
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    • v.14 no.3
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    • pp.219-222
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    • 2008
  • Real time PCR was used to discriminate Colletotrichum acutatum and C. gloeosporioides for analysis of population density. Two primers, caInt2 and cgint, used for conventional PCR to discriminate two species were modified with fluorescent dye to make probe for real time PCR. Fluorescence signals were successfully detected by fCaInt2 and vCgint probe coupled with primer pair Unicon and Unicor1 resulting in discrimination of C. acutatum and C. gloeosporioides by comparison of delta Rn value.

Improved Detection of Viable Escherichia coli O157:H7 in Milk by Using Reverse Transcriptase-PCR

  • Choi, Suk-Ho;Lee, Seung-Bae
    • Food Science of Animal Resources
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    • v.31 no.2
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    • pp.158-165
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    • 2011
  • A sensitive reverse transcriptase-PCR (RT-PCR) method to detect viable Escherichia coli O157:H7 in milk was established. The primer sets were designed based on the nucleotide sequences of the rfbE (per) and wbdN genes in the O157 antigen gene cluster of E. coli O157:H7. RT-PCR using five different primer sets yielded DNA with sizes of 655, 518, 450, and 149-bp, respectively. All five of the E. coli O157:H7 strains were detected by RT-PCR, but 11 other bacterial species were not. The sensitivity of RT-PCR was improved by adding yeast tRNA as a carrier to the crude RNA extract. The RT-PCR amplifying the 149-bp DNA fragment was the most sensitive for detecting E. coli O157:H7 and the most refractory to the bactericidal treatments. Heat treatment at $65^{\circ}C$ for 30 min was the least inhibitory of all bactericidal treatments. Treatment with RNase A strongly inhibited the RT-PCR of heated milk but not unheated milk. This study described RT-PCR methods that are specific and sensitive with a detection limit of 10 E. coli O157:H7 cells, and showed that pre-treating milk samples with RNase A improved the specificity to detect viable bacteria by RT-PCR.

Electrochemical Determination of Bisphenol A at Carbon Nanotube-Doped Titania-Nafion Composite Modified Electrode

  • Kim, Byung Kun;Kim, Ji Yeon;Kim, Dong-Hwan;Choi, Han Nim;Lee, Won-Yong
    • Bulletin of the Korean Chemical Society
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    • v.34 no.4
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    • pp.1065-1069
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    • 2013
  • A highly sensitive electrochemical detection method for bisphenol A (BPA) has been developed by using multi-walled carbon nanotube (CNT)-doped titania-Nafion composite modified glassy carbon (GC) electrode. The CNT-titania-Nafion/GC electrode exhibited excellent electrocatalytic activity towards BPA. Therefore, the CNT-titania-Nafion/GC electrode showed improved voltammetric responses for BPA compared to that obtained with bare GC electrode. In addition, cetyltrimethylammonium bromide (CTAB), a cationic surfactant, was added into the BPA sample solution in order to accumulate BPA through hydrophobic interaction between CTAB and BPA. The CNT-titania-Nafion/GC electrode gave a linear response ($r^2$ = 0.999) for BPA from $1.0{\times}10^{-8}$ M to $5.0{\times}10^{-6}$ M with a detection limit of $9.0{\times}10^{-10}$ M (S/N = 3). The modified electrode showed good selectivity against interfering species and also exhibited good reproducibility. The present electrochemical sensor based on the CNT-titania-Nafion/GC electrode was applied to the determination of BPA in food package samples.