• Journal of Veterinary Clinics
• /
• v.18 no.3
• /
• pp.201-205
• /
• 2001

For evaluating the effect of triple therapy on eradication of gastric Helicobacter species infection in dogs, 7 dogs that had naturally acquired Helicobacter spp. infections were administered amoxicillin, metronidazole and omeparazole orally for 14 days. Changes of infection state were determined by urease test for gastric biopsies and Helicobacter specific PCR analysis for gastric biopsies and fecal samples at 7, 14 days after triple therapy and 30 days after cessation of triple therapy. Although negative results for urease test were obtained 6 of 7 dogs at 14 days after starting triple therapy, PCR analysis for gastric biopsies and fecal samples showed negative results in 3 and 4 dogs respectively. At 30 days after cessation of triple therapy, all tests showed negative results in 3 dogs. Based on these results, diagnostic tests for detercting Helicobacter spp. infection are recommended in dogs having chronic gastritis sign (usually intermittent vomiting) and triple therapy described in this study can be applied for eradicating the organism if the animals were proved to be infected.

• Korean Journal of Microbiology
• /
• v.44 no.3
• /
• pp.212-220
• /
• 2008

A Pn10 DNA probe was introduced as a Prevotella nigrescens ATCC $33563^T$-specific DNA probe. In that study, the specificity of the Pn10 was tested with only type or reference strains of 5 oral bacterial species. The purpose of this study is to evaluate the specificity of the Pn10 using the wild type strains of P. nigrescens and is to develop the P. nigrescens ATCC $33563^T$-specific PCR primers based on the nucleotide sequence of the Pn10. The specificity of the Pn10 DNA probe was determined by Southern blot analysis. The nucleotide sequence of Pn10 DNA probes was determined by chain termination method. The PCR primers were designed based on the nucleotide sequence of cloned DNA fragment. The data showed that Pn10 DNA probe were hybridized with the genomic DNAs from P. nigrescens ATCC $33563^T$ and KB6. The Pn10 homologous region, KB6-Pn10, of P. nigrescens KB6 was cloned by PCR and sequenced. The Pn10 and KB6-Pn10 DNA fragments were consisted of 1,875 bp and 1,873 bp, respectively. The percent identity of the two was 98.8% and the divergence of them was 0.6%. The two primer sets (Pn10-F-AC/ Pn10-R-AC and Pn10-F-A/ Pn10-R-A), designed base on the nucleotide sequences of Pn10 DNA probe, were specific to the P. nigrescens ATCC $33563^T$. The two PCR primer sets could detect as little as 4 pg of genomic DNA of P. nigrescens ATCC $33563^T$. These results indicate that the two PCR primer sets have proven useful for the identification of P. nigrescens ATCC $33563^T$, especially with regard to the maintenance of the strain.

• Journal of the Korean Society of Food Culture
• /
• v.34 no.2
• /
• pp.208-216
• /
• 2019

Leuconostoc spp. are generally utilized as kimchi starters, because these strains are expected to have beneficial effects on kimchi fermentation, including improvement of sensory characteristics. Here, we developed a detection method for verifying the presence of the kimchi starter Leuconostoc mesenteroides WiKim33, which is used for control of kimchi fermentation. A primer set for multiplex polymerase chain reaction was designed based on the nucleotide sequence of the plasmids in strain WiKim33, and their specificity was validated against 45 different strains of Leuconostoc spp. and 30 other strains. Furthermore, the starter strain consistently tested positive, regardless of the presence of other bacterial species in starter kimchi during the fermentation period. Our findings showed that application of a strain-specific primer set for strain WiKim33 presented a rapid, sensitive, and specific method for detection of this kimchi starter strain during natural kimchi fermentation.

• Journal of Microbiology and Biotechnology
• /
• v.25 no.2
• /
• pp.255-267
• /
• 2015

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne's disease, a chronic debilitating disease affecting ruminants worldwide. In the present study, we aimed to determine the major gene networks and pathways underlying the immune response to MAP infection using whole-blood cells, as well as provide the potential transcriptional markers for identifying the status of MAP infection. We analyzed the transcriptional profiles of whole-blood cells of cattle identified and grouped according to the presence of MAP-specific antibodies and the MAP shed by them. The grouping was based on the results obtained by ELISA and PCR analyses as follows: i) Test1 group: MAP-negative results obtained by ELISA and positive results obtained by PCR; ii) Test2 group: MAP-positive results obtained by ELISA and negative results obtained by PCR; iii) Test3 group: MAP-positive results obtained by ELISA and positive results obtained by PCR; iv) uninfected control: MAP-negative results obtained both by ELISA and PCR analysis. The results showed down-regulated production and metabolism of reactive oxygen species in the Test1 group, activation of pathways related to the host-defense response against MAP (LXR/RXR activation and complement system) in the Test2 and Test3 groups, and anti-inflammatory response (activation of IL-10 signaling pathway) only in the Test3 group. Our data indicate a balanced response that serves the immune-limiting mechanism while the host-defense responses are progressing.

• Korean journal of applied entomology
• /
• v.42 no.1
• /
• pp.65-70
• /
• 2003

For the purpose of the protection of beneficial insects from pathogens and the development of control agent against pests, a strain of Metarhizium sp. was isolated from the infected Protaetia brevitarsis seulensis larvae in Korea. Under the scanning electron microscope, the isolate, Metarhizium sp. KMA-1, showed distinct formation of conidia on the palisade-like masse which were comprised of elongate chains and this shape is a typical feature of Metarhizium species. PCR techniques were used to identify the isolate and the primers used were designed on the basis of two kinds of rRNAs sequences, 28S rRNA and internal transcribed spacer(ITS). The specific PCR products from each primer set were amplified and the DNA sequences were determined for the similarity comparison. Sequence alignment of these fragments using GenBank database resulted in the highest homology similarity between the isolate Metarhizium sp. KMA-1 and M. anisopliae. From these results, the isolate Metarhizium sp. KMA-1 in this study was identified as M. anisopliae.

• Food Science and Biotechnology
• /
• v.15 no.6
• /
• pp.967-974
• /
• 2006

Cyclomaltodextrinases (CDases), maitogenic amylases, and neopullulanases share highly conserved primary structures and similar characteristics, and are thus classified into the same family. BLAST search has showed that a variety of bacterial strains harbor putative CDase family genes with several well-conserved motif amino acid sequences. In this study, four degenerate polymerase chain reaction (PCR) primer sets were designed for the detection of CDase genes, on the basis of their highly conserved amino acid blocks (WYQIFP, DGWRLD, LGSHDT, and KCMVW). The PCR detection conditions were optimized and the detection specificity of each for the primer sets was tested against the genomic DNAs isolated from 23 different Bacillus-associated species. Consequently, all tested primer sets evidenced successful amplification of specific PCR products in length, which share 55-98% amino acid sequence identity with known and putative CDases. The primers developed herein, therefore, can be applied for the easy and efficient detection and isolation of CDase family genes for the modification of functional food carbohydrates.

• Research in Plant Disease
• /
• v.13 no.1
• /
• pp.24-29
• /
• 2007

A total of 35 strains of Ralstonia solanacearum isolated from wilted pepper and tomato plants in Korea were analyzed for their genetic diversity by bacteriological, pathological and molecular biological approaches. All the strains were identified as R. solanacearum biovar 4 on the basis of physiological and biochemical tests, and species-specific PCR primers. Pathogenicity of the strains was confirmed by inoculating on 4-week-old pepper and tomato seedlings. Using cluster analysis based on repetitive sequence-based polymerase chain reaction (rep-PCR) genomic fingerprints, R. solanacearum strains isolated from pepper and tomato in Korea divided into 6 groups showing a high degree of genetic diversity at 55% similarity level. The genetic diversify of strains was not significantly correlated with their geographic origins and host plants.

• Research in Plant Disease
• /
• v.15 no.2
• /
• pp.77-82
• /
• 2009

The roots of grapevine in the field in which the crown gall was occurred severely in Chungbuk province were collected and Agrobacterium spp. were isolated from the roots using the selective media. The selected 13 isolates were identified as A. tumefaciens with fatty acid analysis using MIDI system, nucleotide sequence of 16S rDNA, biochemical characteristics, and PCR with the species-specific primers. A. vitis, a pathogen of crown gall disease of grapevine was not isolated from the roots. All of the isolates did not show pathogenicity on the tomato seedlings and the stem and root of grapevine. Eric-PCR showed that DNA band patterns of the root isolates were a little more similar to A. tumefaciens than A. vitis. However, overall similarity between the root isolates and the pathogenic strains of A. tumefaciens and A. vitis was low by rep-PCR. These results suggest that a pathogen causing crown gall in grapevine in Chungbuk province may transmitted through the seedlings rather than via soil or roots.

• Korean Journal of Ichthyology
• /
• v.21 no.2
• /
• pp.129-133
• /
• 2009

Random Amplified Polymorphic DNA (RAPD) analysis was used to investigate the genetic variations of Coreoleuciscus splendidus within and among the West Korea Subdistrict populations (in Han and Geum Rivers) and the South Korea Subdistrict populations (in Seomjin and Nakdong Rivers). Twelve random primers were employed to generate RAPD markers. All primers were produced to identify specific RAPD markers between the West and South Korea Subdistrict populations. Analyses of genetic similarity and distance among the West and South Korea Subdistrict populations of C. splendidus also revealed similar results, with low genetic similarity (0.49~0.53) and high distance value (0.63~0.71). UPGMA dendrogram based on genetic distance was also similar in results. Therefore, the West Korea Subdistrict populations and the South Korea Subdistrict populations vary in genetic structure, and C. splendidus in the South Korea Subdistrict may represent a different species.

• Parasites, Hosts and Diseases
• /
• v.57 no.3
• /
• pp.233-242
• /
• 2019

Detailed description of malaria in low transmission areas is crucial for elimination. The current study aimed to provide a comprehensive description for malaria transmission in Jazan, a low transmission district, southwestern Saudi Arabia. Patients at a tertiary care hospital were recruited in our study between August 2016 and September 2018. Malaria diagnosis was performed through a species-specific nested polymerase chain reaction (nested PCR), microscopy and Paramax-$3^{TM}$ rapid detection test (RDT). Malaria was detected in 30 patients by the PCR, with point prevalence of 10.9%. Of these malaria infections, 80% was imported, 26.6% was asymptomatic and 23.3% was sub-microscopic. Malaria was reported throughout the year, with February/March and September/October peaks. Infection was significantly more in males than in females (P=0.01). Likewise, infections were detected more in febrile than in non-febrile patients (P=0.01). Adult aged 15-24 years, fever and travel were identified as high-risk factors. Malaria was primarily attributed to Plasmodium falciparum mono-infections, followed by P. vivax mono-infections and lastly to falciparum/vivax mixed infections accounting 76.6%, 16.6%, and 6.6% of PCR-confirmed malaria cases, respectively. The nested PCR was superior to the smear microscopy (sensitivity 76.6%; specificity 100%) and the RDT (sensitivity 83.3%, specificity 94.2%). The overall percent agreement between microscopy and the RDT was 92.7% (kappa=0.63). High proportion of imported malaria including sub-microscopic and sub-patent cases were described. We suggest that incorporation of molecular tool into the conventional malaria diagnosis is beneficial in Jazan district.