• Title/Summary/Keyword: Species-specific polymerase chain reaction

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Genetic status of Acanthamoeba spp. Korean isolates on the basis of RAPD markers (RAPD 표지자 분석 에 의한 가시아메바속 한국분리주의 유전적 지위)

  • 홍용표;오승환
    • Parasites, Hosts and Diseases
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    • v.33 no.4
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    • pp.341-348
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    • 1995
  • Genetic status of Acnnthamoebc sap. were tested on the basis of random amplified polymorphic DNA (RAPD) marker analysis. Four previously established Accnthcmoebn species, 4 Korean isolates of Acnnthamoeba sp., and one American isolate of Acanthcmoebc sp. were analyzed by RAPD-PCR using an arbitrary decamer primers. Amplification products were fractionated by agarose gel electrophoresis and slainrd by ethidium bromide . Eighteen primers produced DNA amplification profiles revealing clear differences among 4 species. Nine of them also produced DNA amplification profiles which included some isolate-specific amplification products. On the basis of amplified fragments by 18 primers, the pairwise similarity indices between A. culbensoni and other species (i.e. A. hntchetti, A. trinngularis, A. polyphaga) were 0.300, 0.308, and 0.313, respectively. Similarity index between A. hctchetti and A. triansulcris was 0.833. The mean similarity index among the 3 Korean isolates (YM-2, -3, -4) was 0.959 and 0.832 among them and 2 other species (A. hatchetti and A. triongulnris). The mean similarity index among YM-5 and other Korean isolates (YM-2, -3, -4) was 0.237. However, the similarity index between YM-5 and A. culbeksoni was 0.857, which suggests that YM-5 is genetically more similar to A. culbertsoni than other Korean isolates. Phonogram reconstructed by UPGMA method revealed that there are two groups: one group consists of A. hctchetti, A. tlonsulcns, and 3 Korean isolates (YM-2, -3, -4) , and the other group consists of A. cuLbensoni. A. polwphosc, HOV, and YM-5.

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Molecular Detection of Phellinus linteus and P. baumii by PCR Specific Primer

  • Nam, Byung-Hyouk;Kim, Gi-Young;Park, Hyung-Sik;Lee, Sang-Joon;Lee, Jae-Dong
    • Mycobiology
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    • v.30 no.4
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    • pp.197-201
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    • 2002
  • Specific primer sets based on ribosomal DNA(rDNA) internal transcribed specer(ITS) sequences were designed for rapid detection of Phellinus linteus and P. baumii. Polymerase chain reaction(PCR) with these primers produced unique bands for each Phellinus species. The annealing temperature range is from $40^{\circ}C\;to\;55^{\circ}C$. The length of PCR products(P. linteus and P. baumii) using designed combinative primer sets of PL1F, PL2R, PB1F, PB2R, ITS5F and ITS4R, were from 520 by to 730 bp. Fifteen strains of Phellinus species including P. linteus, P. baumii, P. weirianus, P. johnsonianus, P. rhabarberinus, P. pini, P. gilvus, P. igniarius, P. nigricans and P. laevigatus were examined in this study. Five strains, including two isolated strains of P. linteus(MPNU 7001 and MPNU 7002), and two isolated strains of P. baumii(MPNU 7004 and MPNU 7005) were shown to have about 520 bp (PL1F-PL2R), 700 bp (TTS5F-PL2R) and 600 bp (PB1F-ITS4R) -sized PCR single bands respectively. This molecular genetic technique provided a useful method for rapid detection and identification of P. linteus and P. baumii.

Microbial population dynamics in constructed wetlands: Review of recent advancements for wastewater treatment

  • Rajan, Rajitha J.;Sudarsan, J.S.;Nithiyanantham, S.
    • Environmental Engineering Research
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    • v.24 no.2
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    • pp.181-190
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    • 2019
  • Constructed wetlands are improvised man-made systems, designed for adopting the principle of natural wetlands for purifying wastewater - the elixir of life. They are used widely as a cost-effective and energy-efficient solution for treating greywater generated from different tertiary treatment sources. It provides an elaborate platform for research activities in an attempt to recycle earth's natural resources. Among the several organic impurities removal mechanisms existing in constructed wetland systems, the earth's active microbial population plays a vital role. This review deals with the recent advancements in constructed wetland systems from a microbiological perspective to (effect/ devise/ formulate) chemical and physical treatment for water impurities. It focuses on microbial diversity studies in constructed wetlands, influence of wetland media on microbial diversity and wetland performance, role of specific microbes in water reuse, removal of trace elements, some heavy metals and antibiotics in constructed wetlands. The impurities removal processes in constructed wetlands is achieved by combined interactive systems such as selected plant species, nature of substrate used for microbial diversity and several biogeochemical effected reaction cycles in wetland systems. Therefore, the correlation studies that have been conducted by earlier researchers in microbial diversity in wetlands are addressed herewith.

Ceruloplasmin Gene Expression in U-937 Cells exposed to ${\gamma}$-Irradiation and $H_2$O_2 (U-937 세포에서 방사선 및 $H_2$O_2$에 의한 ceruloplasmin의 mRNA 유전자 발현)

  • 오연경;박선영;김인규;윤병수
    • Environmental Mutagens and Carcinogens
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    • v.22 no.2
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    • pp.76-82
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    • 2002
  • In human U-937 cell exposed to ${\gamma}$-irradiation and $H_2O$$_2$, the level of mRNA efrpression in ceruloplasmin gene was measured by using comparative RT.PCR (reverse transcriptase-polymerase chain reaction). At the normal growth condition, the level of ceruloplasmin transcript was estimated as 8.2% and 0.0068% of hprt (hypoxantine phosphoribosyl transferase) transcript and of $\beta$-actin transcript, respectively. In U-937 cells exposed to a dose of 100 rad ${\gamma}$-irradiation, the level of ceruloplasmin transcript was increased about 2.7 and 1.6 fold compared to un-treated cell by using compensation with the levels of hprt and $\beta$-actin transcript. By contrast, the expression of ceruloplasmin gene in U-937 cells exposed to $H_2O$$_2$(50 $\mu$M, 24 h), was shown no significant difference compared to un-treated cell. These results indicated that the expression system of ceruloplasmin gene may react only some specific oxygen species, such as reactive oxygen species induced by ${\gamma}$-irradiation.

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Identification of Nocardia seriolae by polymerase chain reaction (PCR에 의한 Nocardia seriolae의 검출)

  • Park, Myoung-Ae;Cho, Mi-Young;Kim, Myoung-Sug;Kim, Jae-Hoon;Lee, Deok-Chan
    • Journal of fish pathology
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    • v.22 no.1
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    • pp.85-90
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    • 2009
  • A method for the identification of Nocardia seriolae, the causative agent of nocardiosis in cultured fishes, using PCR was developed in the study. A PCR primer set specific to N. seriolae was designed based on 16S-23S rRNA sequence of various Nocardia species accessed in GenBank. Designed PCR primer set, Nseri-F (5'-GCA AAC TCT TCG AAC AGT CG-3') and Nseri-R (5'-GGA TAT CAG GAC TTA CCG GC-3'), amplifies the target regions of N. seriolae only, but not 4 other Nocardia species, N. asteroides, N. crassostreae, N. farcinica and N. salmonicida.

Screening of Antioxidative Activity of Bifidobacterium Species Isolated from Korean Infant Feces and Their Identification

  • Kim, Ji-Youn;Park, Soo-Im;Heo, Tae-Ryeon
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.3
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    • pp.199-204
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    • 2003
  • Among 59 Korean isolated, 20 were confirmed as members of the genus Bifidobaferium species based on gram staining, microscopic examination of cell morphology and the TLC method, The oxygen tolerance and antioxidative activities of these 20 Bifidobacterium strains and 5 standard Bifidobaferium strains were tested. All the strains demonstrated antioxidative activities as regards inhibiting linoleic acid peroxidation. The antioxidative activities of isolated and standard strains were found to range from 10.7-46.4% and from 10.7-22.2%, respectively. In addition, all tested strains exhibited a Scavenging ability on DPPH free radicals, range from 15-41% for the isolated strains and 8.3-22% for the standard strain. Accordingly. the isolated Bifidobarterium strains demonstrated higher antioxidative artivities than the 5 standa rd Bifidobarterium strains. On the base of grades for each test, HJL 7511 was identified 35 the best strain, followed by HJL 7501. 2 strains were identified with Polymerase Chain Reaction (PCR) assay using group-specific primers designed from the nucleotide Sequences of the 16S rRNA and internal transcribed spacer (ITS) regions of the Bifidobacteria. Based on the Sequencing results, HJL 7511 and HJL 7501 were identified as Bifidobacterium infantis.

Molecular Identification of Cryptosporidium viatorum Infection in a Patient Suffering from Unusual Cryptosporidiosis in West Bengal, India

  • Sardar, Sanjib Kumar;Ghosal, Ajanta;Saito-Nakano, Yumiko;Dutta, Shanta;Nozaki, Tomoyoshi;Ganguly, Sandipan
    • Parasites, Hosts and Diseases
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    • v.59 no.4
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    • pp.409-413
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    • 2021
  • In this study, we have collected and screened a total of 268 stool samples from diarrheal patients admitted to an Infectious disease hospital in Kolkata for the presence of Cryptosporidium spp. The initial diagnosis was carried out by microscopy followed by genus specific polymerase chain reaction assays based on 70 kDa heat shock proteins (HSP70). DNA sequencing of the amplified locus has been employed for determination of genetic diversity of the local isolates. Out of 268 collected samples, 12 (4.48%) were positive for Cryptosporidium spp. Sequences analysis of 70 kDa heat shock proteins locus in 12 Cryptosporidium local isolates revealed that 2.24% and 1.86% of samples were showing 99% to 100% identity with C. parvum and C. hominis. Along with the other 2 major species one recently described globally distributed pathogenic species Cryptosporidium viatorum has been identified. The HSP70 locus sequence of the isolate showed 100% similarity with a previously described isolate of C. viatorum (Accession No. JX978274.1, JX978273.1, and JN846706.1) present in GenBank.

Molecular Identification of Korean Mountain Ginseng Using an Amplification Refractory Mutation System (ARMS)

  • In, Jun-Gyo;Kim, Min-Kyeoung;Lee, Ok-Ran;Kim, Yu-Jin;Lee, Beom-Soo;Kim, Se-Young;Kwon, Woo-Seang;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.34 no.1
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    • pp.41-46
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    • 2010
  • Expensive herbs such as ginseng are always a possible target for fraudulent labeling. New mountain ginseng strains have occasionally been found deep within mountain areas and commercially traded at exorbitant prices. However, until now, no scientific basis has existed to distinguish such ginseng from commonly cultivated ginseng species other than by virtue of being found within deep mountain areas. Polymerase chain reaction (PCR) analysis of the internal transcribed spacer has been shown to be an appropriate method for the identification of the most popular species (Panax ginseng) in the Panax ginseng genus. A single nucleotide polymorphism (SNP) has been identified between three newly found mountain ginseng (KGD4, KGD5, and KW1) and already established Panax species. Specific PCR primers were designed from this SNP site within the sequence data and used to detect the mountain ginseng strains via multiplex PCR. The established multiplex-PCR method for the simultaneous detection of newly found mountain ginseng strains, Korean ginseng, and foreign ginseng in a single reaction was determined to be effective. This study is the first report of scientific discrimination of "mountain ginsengs" and describes an effective method of identification for fraud prevention and for uncovering the possible presence of other, cheaper ginseng species on the market.

Molecular Detection and Analysis of Sweet potato feathery motile vims from Root and Leaf Tissues of Cultivated Sweet Potato Plants

  • Ryu, Ki-Hyun;Park, Sun-Hee
    • The Plant Pathology Journal
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    • v.18 no.1
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    • pp.12-17
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    • 2002
  • For the molecular detection of Sweet potaio feathery mottle virus (SPFMV) from diseased sweet potato plants, reverse transcription and polymerase chain reaction (RT-PCR) was performed with the use of a set of virus-specific primers to amplify an 816 bp product. The viral coat protein gene was selected for the design of the primers. No PCR product was amplified when Turnip mosaic virus, Potato vims Y or Cucumber mosaic virus were used as template in RT-PCR with the SPFMV-specific primers. The lowest concentration of template viral RNA required for detection was 10 fg. The vim was rapidly detected from total nucleic acids of leaves and roots from the virus-infected sweet potato plants as well as from the purified viral RNA by the RT-PCR. Twenty-four sweet potato samples were selected and analyzed by RT-PCR and restriction fragment length polymorphism (RFLP). RFLP analysis of the PCR products showed three restriction patterns, which resulted in some point mutations suggesting the existence of quasi-species for the vims in the infected sweet potato plants.

Rapid PCR Method for Detecting Candida albicans Using Primers Derived from the Integrin-like Protein Gene $\alpha$INT1 of Candida albicans

  • Lim, Young-Hee;Lee, Do-Hyun
    • Journal of Microbiology
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    • v.38 no.2
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    • pp.105-108
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    • 2000
  • Oligonucleotide primers amplifying a 344 bp fragment on the integrin-like protein alpha-INT1p gene (${\alpha}$INT1) of Candida albicans were synthesized for screenign of C. albicans from clinicalsamples by the polymerase chain reaction (PCR). The PCR specifically amplified DNA from C. albicans and none from any other Candida, fungal, or human DNA in standard used here. The PCR assay showed that the primers (LH1 and LH2) were specific for 26 isolates of C. albicans from clinical smaples, whereas the positive fragment, 344 bp, was not amplified from 15 clinical isolates including 14 other medically important Candida species and an isolate of Saccharomyces cerevisiae. PCR was conducted on the urine samples of 20 patients and 4 samples were C. albicans positive. The detection limit of the PCR assay for C. albicans was shown to be approximately 10 cells/ml saline. The PCR system using 344 bp ${\alpha}$INT1 as a target is more specific and rapid than the conventional culture method, and the sensitive detection method is applicable to clinical diagnosis of C. albicans infections.

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