• Korean Journal of Clinical Laboratory Science
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• v.49 no.4
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• pp.413-419
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• 2017

Four imipenem-resistant bacteria were isolated from the clinical specimens of a patient with pneumonia. To identify the isolates, we used the GN card of Vitek II system and performed a phylogenetic analysis based on 16S rRNA gene sequence. The isolates were identified as P. aeruginosa (2 strains), P. monteilii (1 strain), and P. putida (1 strain), and were tested for antibiotic resistance after determining the MIC of imipenem to be $${\geq_-}8{\mu}g/mL$$ using the AST-N225 card of Vitek II system. The imipenem-resistant genotypes were determined using PCR products amplified using specific ${\beta}-Lactamase$ gene primers. The MBL gene was identified in all four isolates. One strain of P. aeruginosa exhibited the VIM and SHV-1 type genes, while the other strain exhibited both VIM and OXA group II genes. According to the antimicrobial susceptibility test, the bacteria were more susceptible to amikacin than other antibiotics. DNA fingerprint analysis using ERIC-PCR to analyze the epidemiological relationship between strains estimated that both the P. aeruginosa isolates were similar, but exhibited different DNA band types. It is uncommon to find four strains of imipenem-resistant bacteria with different DNA band types in a single patient.

• The Plant Pathology Journal
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• v.27 no.1
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• pp.33-36
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• 2011

The genome of a potyvirus isolate associated with chlorotic spots and vein banding symptoms on Spathiphyllum spp., in Andhra Pradesh state, India was amplified by RT-PCR using degenerate potyvirus primers, amplicons cloned, and sequence (1.6 kb) analyzed. This virus isolate shared maximum identity of 74.8% and 80.2% at coat protein (CP) gene nucleotide (906 nucleotides) and amino acid (302 amino acids) levels, respectively with Dasheen mosaic virus (DsMV)-M13 isolate reported from China. But its 3'-UTR (258 nucleotides) had maximum identity of 62.5% with DsMV-Vietnam isolate. The deduced molecular weight of CP is 33.57 kDa and it contained DAG triplet in its N-terminal region. In CP amino acid based phylogenetic analysis, this virus isolate represented a separate branch but closer to DsMV isolates cluster. Based on the molecular criteria set for the discrimination of species and genus in the Potyviridae family, the present virus isolate was identified as a distinct virus species in the genus Potyvirus and proposed the name Spathiphyllum chlorotic vein banding virus (SCVbV).

• Korean Journal of Veterinary Research
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• v.40 no.2
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• pp.301-310
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• 2000

The antimicrobial susceptibility, genotypes of enterotoxins(LT, STa) and pili(K88, 987P), and plasmid profiles were investigated with 102 Escherichia coli isolated from piglets showing diarrhea in Korea. Almost of them were susceptible to ceftiofur(99%), cefquinone(97.1%). However they showed resistance to bacitracin(100%), streptomycin(98%), vancomycin(97%), trimethoprim/sulfamethoxazole(87.2%), tetracycline(84.3%) in antimicrobial susceptibility test. Moreover, all of the isolates demonstrated resistance to more than 2, and 78% of them were resistant to more than 8 of total 17 drugs. Multiplex PCRs for genotyping of enterotoxins(LT, STa) and pili(K88, 987P) were established with primers designed based on sequences from Genebank. Seventeen strains(16.6%) of the isolates had STa gene, 11 strains(10.8%) of them had both STa and LT genes, and 18 strains(17.8%) had K88 gene. But none of the isolates harbored a gene exclusively encoding LT. The gene encoding 987P pili was not found in all isolates. Fifty-four strains of 102 isolates(52.9%) had plasmid with various sizes ranging from 125kb to 1.1kb. Numbers of plasmid per isolates were also various, from 1 to 9. Distinctive relationship between plasmid profiles and genotypes of enterotoxins and pili in the isolates was not found. These results might provide the basic knowledge to establish strategies for the treatment and prevention of colibacillosis in piglets.

• Mycobiology
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• v.38 no.1
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• pp.17-25
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• 2010

The common split-gilled mushroom, Schizophyllum commune is found throughout the world on woody plants. This study was initiated to evaluate conditions for favorable vegetative growth and to determine molecular phylogenetic relationship in twelve different strains of S. commune. A suitable temperature for mycelial growth was obtained at $30^{\circ}C$. This mushroom grew well in acidic conditions and pH 5 was the most favorable. Hamada, glucose peptone, Hennerberg, potato dextrose agar and yeast malt extract were favorable media for growing mycelia, while Lilly and glucose tryptone were unfavorable. Dextrin was the best and lactose was the less effective carbon source. The most suitable nitrogen sources were calcium nitrate, glycine, and potassium nitrate, whereas ammonium phosphate and histidine were the least effective for the mycelial growth of S. commune. The genetic diversity of each strain was investigated in order to identify them. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 129 to 143 bp and 241 to 243 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical. A phylogenetic tree of the ITS region sequences indicated that the selected strains were classified into three clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) with 20 arbitrary primers. Twelve primers efficiently amplified the genomic DNA. The number of amplified bands varied depending on the primers used or the strains tested. The average number of polymorphic bands observed per primer was 4.5. The size of polymorphic fragments was obtained in the range of 0.2 to 2.3 kb. These results indicate that the RAPD technique is well suited for detecting the genetic diversity in the S. commune strains tested.

• The Journal of Korean Academy of Prosthodontics
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• v.34 no.4
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• pp.833-849
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• 1996

The material of choice for functional and esthetic reconstruction of maxillofacial defects is silicone. Silicone has appropriate physical properties for maxillofacial prosthesis but it has weak edge strength. Therefore, a proper combination of silicone and polyurethane sheet is recommended to improve this weakness. Various primers are also used to enhance the adhesive strength between silicone and polyurethane sheet. The purpose of this study was to determine the adhesive strength of silicone and polyurethane sheet. Silicone elastomer mixture was made by admixing MDX4-4210 elastomer (40%) and Silastic Medical Adhesive Type A(60%). This silicone elastomer mixture was attached to polyurethane sheet, using one of three different primers(1205, S-2260, or A-304), treated for 1, 2, 4, 6, and 8 hours. These were then polymerized in room temperature, dry-heat oven or microwave oven. Six specimens per each group, a total of 270 specimens were prepared for final test. The differences of T-peel bonding strengths were then determined by a test. The differences of T-peel bonding strengths were then determined by a test method that was recommended by American Society for Testing and Materials C794-80. The results were statistically analyzed using the ANOVA and Mutiple Range Tests(Tukey' HSD). The reults were as follow. 1. Type of primer, primer reaction time, and methods of polymerization showed significant correlation on the T-peel bonding strengths in adhesiveness between silicone and polyurethane sheet. 2. A-304 primer showed statistically higher in T-peel bonding strength than otehr type of primers except for the polymerization in microwave oven with reaction times of 2, 6 hours(p<0.05). 3. No significant differences in T-peel bonding strength were observed among the polymerization methods. 4. The effect of reaction time by the primer type and polymerization method showed statistically significant differences in bonding strength among different reaction times. And in most cases, reaction time of 1 or 2 hours showed higher T-peel bonding strength.

• Journal of Microbiology and Biotechnology
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• v.19 no.8
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• pp.749-759
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• 2009

A survey of Bacillus thuringiensis (Berliner) strains isolated from Spanish citrus orchards has been performed, and the strains were tested for insecticidal activity against the Mediterranean fruit fly Ceratitis capitata (Wiedemann), a key citrus pest in Spain. From a total of 150 environmental samples, 376 isolates were selected, recording a total B. thuringiensis index of 0.52. The collection was characterized by means of phase-contrast microscopy, SDS-PAGE, and PCR analysis with primer pairs detecting toxin genes cry1, cry2, cry3, cry4, cry5, cry7, cry8, cry9, cry10, cry11, cry12, cry14, cry17, cry19, cry21, cry27, cry39, cry44, cyt1, and cyt2. Diverse crystal inclusion morphologies were identified: bipyramidal (45%), round (40%), adhered to the spore (7%), small (5%), and irregular (3%). SDS-PAGE of spore-crystal preparations revealed 39 different electrophoresis patterns. All primer pairs used in PCR tests gave positive amplifications in strains of our collection, except for primers for detection of cry3, cry19, cry39, or cry44 genes. Strains containing cry1, cry2, cry4, and cry27 genes were the most abundant (48.7%, 46%, 11.2%, and 8.2% of the strains, respectively). Ten different genetic profiles were found, although a total of 109 strains did not amplify with the set of primers used. Screening for toxicity against C. capitata adults was performed using both spore-crystal and soluble fractions. Mortality levels were less than 30%. We have developed a large and diverse B. thuringiensis strain collection with huge potential to control several agricultural pests; however, further research is needed to find out Bt strains active against C. capitata.

• The Korean Journal of Mycology
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• v.24 no.2 s.77
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• pp.155-165
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• 1996

This study was carried out to identify the phylogenetic relationship among several isolates of Pleurotus species by comparing ITS II region of ribosomal DNA(rDNA) repeat unit. Two primers from ribosomal DNA sequences were chosen to amplify the specific internal transcribed spacer (ITS) II region of Pleurotus spp. The exact ITS II region with an unique band from six species of Pleurotus genus could be amplified using the two primers taken from at the 3'-end of 5.8S rDNA and 5'-end of 28S rDNA. Six representative species of the Pleurotus genus were easily characterized according to the length differences of ITS II region. Furthermore, within P. ostreatus species, different sizes of ITS II region could be observed in the isolates of ASI 2025 and ASI 2095 although they were classified as P. ostreatus by the conventional observation. The nucleotide sequence analyses of PCR-amplified ITS II region indicated that the isolates ASI 2025 and ASI 2095 were different from other Pleurotus spp. When the nucleotide sequences of six Pleurotus species were compared, three typical ITS II regions were highly variable especially at both ends of this region. The phylogenetic tree obtained by the Neighbor program of Felsenstein PHYLIP package with all the nucleotide sequence of Pleurotus spp. indicated that P. ostreatus, P. florida, P. sajor-caju and P. eryngii were closely related to one phylogenetic branch and P. cystidious was related to other branch with P. cornucopiae. The isolates ASI 2025 and 2038, however, were not closely related to any other Pleurotus spp. and formed their own individual branches.

• Journal of Microbiology
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• v.44 no.1
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• pp.92-97
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• 2006

We developed an mPCR assay for the simultaneous detection, in one tube, of Escherichia coli O157:H7, Salmonella spp., Staphylococcus aureus and Listeria monocytogenes using species-specific primers. The mPCR employed the E. coli O157:H7 specific primer Stx2A, Salmonella spp. specific primer Its, S. aureus specific primer Cap8A-B and L. monocytogenes specific primer Hly. Amplification with these primers produced products of 553, 312, 405 and 210 bp, respectively. All PCR products were easily detected by agarose gel electrophoresis, and the sequences of the specific amplicons assessed. Potential pathogenic bacteria, in laboratory-prepared and four commercially available kimchi products, were using this mPCR assay, and the amplicons cloned and sequenced. The results correlated exactly with sequences derived for amplicons obtained during preliminry tests with known organisms. The sensitivity of the assay was determined for the purified pathogen DNAs from four strains. The mPCR detected pathogen DNA at concentrations ranging from approximately 0.45 to $0.05\;pM/{\mu}l$. Thus, this mPCR assay may allow for the rapid, reliable and cost-effective identification of four potentially pathogens present in the mixed bacterial communities of commercially available kimchi.

• Journal of Photoscience
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• v.7 no.2
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• pp.73-78
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• 2000

To analyze molecular traits determining pigmentation between Pharbitis nill violet and white, Amplified Fragment Length Polymorphism(AFLP) and Differential Display Reverse Transcription(DDRT) experiments were carried out with either genomic DNAs or total RNAs isolated from both plants. Results of AFLP experiment in combination of 8 EcoRⅠ primers with 6 MseⅠ primers showed 41 violet-and 60 white-specific DNA bands. In the subsequent experiment, 22 violet-and 22 white-specific DNA fragments were amplified by PCR with DNAs eluted. The sizes of the fragments range from 200 to 600bp. DDRT using total RNA produced 19 violet-and 17 white-specific cDNA fragments, ranging from 200 to 600bp. The fragments obtained by both AFLP and DDRT had been cloned into pGEM T-easy vector, amplified and subjected to the nucleotide sequence analyses. As a result of Blast sequence analysis, most of them sequenced up to date showed no similarity to any Known gene, while few has similarity to known animal or plant genes. An AFLP clone V6, for example, has a strong sequence similarity to the human transcription factor LZIP-alpha mRNA and a DDRT clone W19 to Solanum tuberosum 3-hydroxy-3-methylglutaryl coenzyme A reductase mRNA.

• Journal of Mushroom
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• v.13 no.1
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• pp.37-40
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• 2015

Genetic relationship analysis by PCR reactions with 25 primers could produce relevant classifications of the 21 P. pulmonarius (Fr) Quel. isolates collected from 2005. They were classified by two groups with similarity coefficient 0.37. All of the P. pulmonarius cultivars belonged to group 2. New cultivar 'Hwasan' presented high similarity coefficient to parental strains, and that of 'Hosan' and 'Kangsan' cultivars was very high owing to shared crossing parents, and that of two strains from China was high, as well.