• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.9 no.1
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• pp.1-13
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• 2006

Purpose: Acute gastroenteritis in children is one of the frequently encountered diseases with relatively high admission rate. The aim of this study is to determine the isolation trends of common and emerging pathogens in acute gastroenteritis in children over a 12-month period in a community hospital. Methods: The study group included the children who were hospitalized to Seoul National University Boramae Hospital from April, 2003 to March, 2004 or visited outpatient clinic from April, 2003 to July, 2003 with presenting features of acute gastroenteritis. Stool specimens were obtained within 2 days after the visit and examined for the following pathogens: rotavirus, adenovirus, Salmonella, Shigella, Vibrio, pathogenic Escherichia coli (E.coli), Campylobacter and Yersinia species. Viral study was done with commercial kits for antigen detection. Identification of the bacterial pathogens was done by culture using selective media. For pathogenic E.coli, polymerase chain reaction (PCR) was done with the target genes related to the pathogenecity of enterotoxigenic E.coli (ETEC), enteropathogenic E.coli (EPEC) and enterohemorrhagic E.coli (EHEC). Results: The 130 hospitalized children and 28 outpatients were included in this study. The majority of children (>93%) were less than 6 years. Pathogens were isolated in 47% of inpatients and 43% of outpatients, respectively. Rotavirus was the most frequently identified pathogen, accounting for 42.3% of inpatients and 29.6% of outpatients. Nontyphoidal salmonella is the most commonly isolated bacterial pathogen (3.9%) in hospitalized children. Pathogenic E.coli (EPEC, ETEC) was detected in 2.1% (2/97) of inpatients and 25% (3/12) of outpatients. EHEC, adenovirus, Campylobacter, Yersinia and Shigella species were not detected in this study. Conclusion: Rotavirus is the most common enteropathogen in children with acute gastroenteritis. Nontyphoidal salmonella and pathogenic E.coli are important bacterial pathogens. Campylobacter species may not be commonly detected organism in hospitalized children with acute diarrhea.

• Pediatric Infection and Vaccine
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• v.18 no.2
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• pp.154-162
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• 2011

Purpose : The aim of this study was to identify the clinical characteristics of lower respiratory tract infection due to respiratory syncytial virus (RSV) in young children and to provide information for an effective guideline for palivizumab administration in Korea. Methods : We reviewed medical charts of 167 patients under 3 years of age who were hospitalized in Dongguk University Ilsan Hospital for lower respiratory tract infection between January 2007 and February 2011. Diagnosis of the virus was made based on the multiplex real time polymerase chain reaction. Results : There were 113 patients who were infected by respiratory syncytial virus. 90 patients were term infants and 23 patients were preterm infants. No difference was shown between term and preterm infants except the days of admission which was 9.0${\pm}$6.0 days and 12.6${\pm}$21.0 days respectively. In the preterm group their mean age at the time of admission was 5.21${\pm}$4.9 months and the mean gestational age was 33.1${\pm}$4.3 weeks, and the mean birth weight was 2,152${\pm}$950 g. Only 4 patients were born under 28 weeks gestational age and were candidates for palivizumab administration. Conclusion : Most of the patients with severe RSV lower respiratory tract infection were term or near term infants who were not candidates for palivizumab prophylaxis. A nationwide study is needed to make a new risk stratified guideline for RSV prophylaxis for our country.

• Pediatric Infection and Vaccine
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• v.16 no.1
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• pp.31-39
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• 2009

Purpose : Neisseria meningitides is one of the most common causative pathogens of bacteremia and meningitis. Recently protein-conjugated vaccines have been developed and included in the routine vaccination schedule in a few countries. In Korea, carriage rates of N. meningitides among healthy adults have been reported. However, systematic data for childhood carriage rates are not available. This study was performed to evaluate the carriage rates of N. meningitides and the serotype distribution among healthy children attending day care centers. Methods : During the period of January through May 2005, nasopharyngeal swabs and culture were obtained from 904 children attending 13 different day care centers located in Seoul and Gyeonggi Province. The Vitek NHI card was used to identify N. meningitides and the crgA gene was detected via polymerase chain reaction (PCR). Serotype determination was performed by agglutination test using N. meningitides antisera to serotypes A, B, C, D, 29E, W135, X, Y, and Z. PCR for detection of the org2 and saiD gene confirmed serotypes A, B, C, W135, and Y. Results : The mean age among 904 children was 4.5 years; 6.5% (59/904) were children <2 years old, 53.8% (486/904) were 2-5 years old, and 39.7% (359/904) were >5 years old; 52.0% (468/904) were male. N. meningitides was isolated from only 7 children attending 5 different day care centers and the overall carriage rate of N. meningitides was 0.8%. The detected serotypes of N. meningitides were serotype A (n=2), C (n=2), and Y (n=3). Conclusion : The carriage rate of N. meningitides among healthy children attending day care centers was very low in Korea and the detected serotypes were A, C, and Y.

• Journal of Life Science
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• v.17 no.12
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• pp.1641-1648
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• 2007

Yippee-like proteins, which have been identified as the homolog of Drosophila yippee protein containing a zinc-finger domain, are known to be highly conserved among eukaryotes. However, their functional roles are still poorly understood. Recently we initiated ordered differential display (ODD)-polymerase chain reaction (PCR) to isolate genes of which expressions are altered following activation of human T cells. On the ODD-PCR image, one PCR-product detected in unstimulated T cells was not detectable at the time when the activated T cells traversed near $G_1/S$ boundary following activation by immobilized anti-CD3. Cloning and nucleotide sequence analysis revealed that the PCR-product was yippee-like 5 (YPEL5) gene, which was known as a human homolog of the Drosophila yippee gene. Northern blot analysis confirmed the amount of ${\sim}2.2$ kb YPEL5 mRNA expression detectable in unstimulated T cells was sustained until 1.5 hr after activation and then rapidly declined to undetectable level by 5 hr. Ectopic expression of YPEL5 gene in human cervix epitheloid carcinoma HeLa cells caused a significant reduction in cell proliferation to the level of 47% of the control. Expression of GFP-YPEL5 fusion protein in HeLa cells showed its nuclear localization. These results demonstrated that the expression level of human YPEL5 mRNA was negatively regulated in the early stage of T cell activation, and suggested that YPEL5 might exert an inhibitory effect on the cell proliferation as a nuclear protein.

• Journal of Life Science
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• v.17 no.3 s.83
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• pp.386-395
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• 2007

A total of 27 strains of Vibrio parahaemolyticus (18 strains isolated from Korea and 9 strains from Japan) were serotyped and examined for biochemical characteristics, antimicrobial susceptibility patterns, cytotoxicity assay, thermostable direct hemolysin (TDH) production and molecular epidemiology. Using polymerase chain reaction (PCR) method and DNA probe hybridization method, the strains were tested for toxR, tdh, trh and ORF 8 genes. The V. parahaemolyticus isolated from patients were belonged to 8 different serotypes : O3:K6, O1:K38, O3:K57, O4:K9, O4:Kl2, O4:K68, O5:Kl5 and O6:K46. Urease-positive strain possessed the trh gene, and conversely, urease-negative strains lacked the gene, indicating that urease production by V. parahemolyticus strains strongly correlates with the possession of the trh gene. Most strains showed multiple resistant to more than three antibiotics and the antibiogram could be classified into 6 group (I to VI). All of the O3:K6 strains isolated in South Korea and Japan producted TDH at high levels. The TDH titers ranged between 256 and 2.048, and the average titer was 1009. To distinguish the new and increasingly common V. parahaemolyticus strains from clinical isolates, ORF 8 is a useful genetic marker. After Southern hybridization, the HindIII restriction fragment patterns of the tdh gene were grouped one type, respectively. One type showed two bands one of which was 4.3kb and the other was 11.5kb in size. Variation between the O3:K6 serotype are minor when compared to the differences seen with the non O3:K6 strains. The migration patterns of Not I -digested of the total DNA of the O3:K6 strains were similar, and only slight variations were observed between the serotypes. By contrast, the O3:K6 strains and non O3:K6 had markedly different profiles. In conclusion, Random amplified polymorphic DNA (RAPD) profile using appropriate primers was an effective epidemiological marker.

• Korean Journal of Food Science and Technology
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• v.45 no.1
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• pp.1-7
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• 2013

Niphon spinosus, Epinephelus bruneus, and Epinephelus septemfasciatus are involved in the Perciformes Order and Serranidae Family. When E. bruneus and E. septemfasciatus are fully grown, the striped pattern on the body gradually disappears. Therefore, morphological classification of adult fishes is quite difficult to identify the differences to N. spinosus. In this study, we investigate the method to differentiate those using PCR. To design the primers, 16S rRNA region of N. spinosus, E. bruneus, and E. septemfasciatus registered in the GeneBank (www.ncbi.nlm.nih.gov) have been used and for the analysis, Bio Edit ver. 7.0.9.0 was used. As a result, it was design NS-003-F/NS-005-R (136 bp), EB-001-F/EB-002-R (181 bp), and ES-001-F/ES-001-R (123 bp) primers for the differentiation of each 3 different fishes. Therefore, the species-specific primer sets would be a useful tool for scientific and speedy differentiation against the illegal distribution for consumer protection.

• Journal of Food Hygiene and Safety
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• v.30 no.3
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• pp.281-288
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• 2015

Vibrio is a genus of Gram-negative, curved, halophilic, and non-spore-forming bacteria. Some of the Vibrio species, such as V. cholerae and V. parahaemolyticus, often contaminate seafood products and occasionally cause human diseases when the seafood products are ingested. A total of 24 Vibrio strains were isolated from shrimp samples on Thiosulphate citrate bile salt sucrose (TCBS) media in this study. All of the 24 isolates were confirmed to belong to the genus Vibrio by using 16S rRNA gene sequence analyses. Vitek 2 system and species-specific polymerase chain reaction (PCR) methods were used to further identify a total of 29 Vibrio strains at the species level, including the 24 shrimp Vibrio isolates and five Vibrio reference strains. The specificities of the two methods to identify Vibrio strains at the species level were compared in this study. The species-specific PCR method was designed to detect five different Vibrio species, such as Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio alginolyticus, and Vibrio mimicus. From the 24 Vibrio shrimp isolates, the Vitek 2 system method could identify 15 (62.5%) strains as Vibrio species and 7 (29.2%) strains as non-Vibrio species, but could not identify the rest 2 (8.3%) strains. But species-specific PCR method could identify 16 (66.7%) strains as Vibrio species and could not identify the rest 8 (33.3%) strains. Among the 24 Vibrio shrimp strains, these two methods could unanimously identify 7 (7/24, 29.2%) strains (2 V. parahaemolyticus, 4 V. alginolyticus, and 1 V. mimicus). Considering that such different identification results were obtained using the two different methods in this study, identification method for Vibrio species must be carefully chosen.

• Journal of Food Hygiene and Safety
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• v.29 no.4
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• pp.340-346
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• 2014

In this study, the detection method was developed using real-time PCR to distinguish 4 species (bovine, porcine, horse, and chicken) of raw meats. The genes for distinction of species about meats targeted at 12S rRNA and 16S rRNA parts in mitochondrial DNA. Probes were designed to have a 5' FAM and a TAMRA at the 3' end. This study is to develop 4 species-specific primer and probes about raw materials and real-time PCR on 10 strains to observe the products of non-specific signal for similar species. As a result, any non-specific signal were not detected among each other. Real-time PCR method was developed for quantitation and identification of intentional and unintentional mixture in ground mixed meat (The difference of $C_T$ value between intentional mixture and 100% meat: $${\leq_-}$$ cycles, The difference of $C_T$ value between unintentional mixture and 100% meat: $${\geq_-}$$ cycles). The detection and differentiation of intentional and unintentional mixture in this study would be applied to food safety management for eradication of adulterated food distribution and protection of consumer's right.

• Annals of Clinical Microbiology
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• v.21 no.4
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• pp.86-91
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• 2018

Background: The PANA RealTyper HPV kit (PANAGENE, Korea; PANA RealTyper) was developed to genotype human papillomavirus (HPV) and was based on multiplex real-time PCR amplification and melting curve analysis. In this study, we compared PANA RealTyper to the AdvanSure HPV GenoBlot assay (LG Life Sciences, Korea; AdvanSure assay) and attempted to evaluate the performance of PANA RealTyper. Methods: A total of 60 cervical specimens were collected from women undergoing routine cervical cancer screening. The AdvanSure assay and PANA RealTyper kit identified the same 20 high-risk genotypes. However, the AdvanSure assay identified 15 low-risk genotypes, while the PANA RealTyper kit identified only 2 but detected 18 low-risk genotypes. Results: Among the total 60 specimens, 54 high-risk genotypes (40 specimens) and 20 low-risk genotypes (18 specimens) were detected. The agreement rates of the assays ranged from 94.4 to 100% for high-risk genotypes. Among 9 genotypes that were positive in the PANA RealTyper kit but negative in the AdvanSure assay, 7 were confirmed as true positive (HPV genotypes 16 (n=1), 39 (n=1), 52 (n=1), 58 (n=2), 68 (n=2)). Among 4 genotypes that were negative in the PANA RealTyper kit but positive in the AdvanSure assay, 3 were confirmed as HPV genotype 59. Among the 19 low-risk genotypes positive in the AdvanSure assay, there were 2 cases of HPV 6 and 1 case of HPV 11. In comparison, only 1 positive case of HPV 6 was determined by the PANA RealTyper kit. Conclusion: The PANA RealTyper kit was comparable with the AdvanSure assay. The PANA RealTyper kit would be useful and suitable for HPV genotyping in the clinical laboratory.

• Journal of Sericultural and Entomological Science
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• v.53 no.2
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• pp.135-142
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• 2015

The American cockroaches, Periplaneta americana L. was the most important worldwide pest species. It has been an public health problems. We were determinated life cycle and extraction of crude extracts by chemical reagents from cockraches (P. americana L.). The extracted crude solution has been antibacterial activity to gram negative bacteria (Pseudomonas aeruginosa, $6.44{\pm}1.03mm$), gram positive bacteria (Bacillus subtilis, $1.88{\pm}0.40mm$), and fungus (Candida albicans, $5.61{\pm}0.57mm$) using radial diffusion assay. We were analysed of up-regulation of Glutathione-S-transferases (GSTs) stimulation, indicating that antioxidantial protein from various classes are simultaneously expressed in a single insect upon infection or injury. The gene from Periplaneta americana L. were cloned, analysed sequence, and measured protein expression by Real Time PCR (Polymerase Chain Reaction).