• Journal of Life Science
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• v.24 no.4
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• pp.361-369
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• 2014

Broad-range and specific 16S rRNA gene PCR is used for detection and identification of bacterial pathogens in clinical specimens from patients with a high suspicion for infection. We describe the development of a broad-range and specific PCR primer, based on bacterial 16S rRNA, for use in routine diagnostic clinical microbiology services. The primers were designed by using conservative regions of 16S rRNA sequences from 10 strains. Ninety-eight clinical strains were isolated from clinical patient specimens. A total of 98 strains of bacteria were identified by phenotypic methods; PCR with newly designed primers and universal primers. All purified PCR products were sequenced using both forward and reverse primers on an automated DNA analyzer. In this study, we evaluated the usefulness of the newly designed primers and the universal primers for the detection of bacteria, and both these techniques were compared with phenotypic methods for bacteria detection. When we also tested 98 strains of clinical isolates with newly designed primers, about 778 bp DNA fragments were amplified and identified from all strains. Of the 98 strains, 94 strains (95.9%) correspond in comparison with phenotypic methods. The newly designed primers showed that the identities of 98 (100%) strains were the same as those obtained by universal PCR primers. The overall agreement between the newly designed primers and universal primers was 100%. The primer set was designed for rapid, accurate, and cheap identification of bacterial pathogens. We think the newly designed primer set is useful for the identification of pathogenic bacteria.

• Journal of The Korean Society of Inherited Metabolic disease
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• v.12 no.1
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• pp.23-34
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• 2012

Purpose: Acute symptomatic seizures are caused by structural changes, inflammation or metabolic changes of brain, such as tumor, stroke, meningitis, encephalitis and metabolic disorders. Inherited metabolic disorders that can cause seizures are organic acidopathies, lysosomal storage disorders, peroxisomal disorders and mineral disorders. We have done this study to find out the importance of organic acidopathies causing seizure disorders in Korean childhood and adolescent patients. Method: Retrograde analysis for 1,306 patients with seizure disorders whose clinical informations are available and have done urine organic acid analysis for 5 years period, between Jan. 1st 2007 to Dec. 31th 2011. Statistical analysis was done with Student's t test using SPSS. Result: Out of 1,306 patients, 665 patients (51%) showed abnormalities on urine organic acid analysis. The most frequent disease was mitochondrial respiratory chain disorders (394, 30.1%), followed by mandelic aciduria (127, 9.7%), ketolytic defects (81, 6.2%), 3-hydroxyisobutyric aciduria (19, 1.4%), glutaric aciduria type II (10, 0.8%), ethylmalonic aciduria (4), propionic aciduria (4), methylmalonic aciduria (3), glutaric aciduria type I (3), pyruvate dehydrogenase deficiency (3), pyruvate carboxylase deficiency (3), isovaleric aciduria (2), HMG-CoA lyase deficiency (2), 3-methylcrotonylglycinuria (2), fatty acid oxidation disorders (2), fumaric aciduria (1), citrullinemia (1), CPS deficiency (1), MCAD deficiency (1). Conclusion: On neonatal period, mandelic aciduria due to infection was found relatively frequently. Mitochondrial disorders are most frequent etiologic disease on all age group, followed by ketolytic defects and various organic acidopathies. The number and diversities of organic acidopathies emphasize meticulous evaluation of basic routine laboratory examinations and organic acid analysis with initial sample on every seizure patient.

• Korean Journal of Clinical Laboratory Science
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• v.39 no.2
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• pp.113-121
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• 2007

Hearing loss is a common congenital disorder that is frequently associated with mutations in the Cx26 gene (GJB2). Recently, the mutation analysis of GJB2 has been used in a newborn screening test for the detection of hearing impairment. Population-based studies should be performed before the application of genetic testing for the identification of deaf newborns. In this study, 8 positions of GJB2 mutations-including 35delG, 167delT, 235delC, V27I, V37I, M34T, E114G, and I203T-were analyzed using PCR-direct sequencing in a total of 437 healthy Korean neonates. DNAs from dried blood spots were extracted using a commercial DNA extraction kit. The PCR-amplified products (783 bps) of the GJB2 gene were detected using 2% agarose gel electrophoresis and subjected to direct sequencing. The sequences were compared with those in the GenBank database by using the BLAST program. In this study, 5 GJB2 mutations -including V27I (79G>A), V37I (109G>A), E114G (341A>G), I203T (608T>C), and 235delC- were found. Of the 437 neonate samples, 301 subjects showed GJB2 mutations (68.9%, 301/437). The V27I mutation was found in 271 subjects and was the most frequent (62.0%, 271/437). The E114G, I203T and V37I mutations were shown in 146, 17 and 14 subjects, respectively. The 235delC mutation was found in 1 subject. The E114G mutation was frequently accompanied by the V27I mutation. V27I/E114G (97.2%, 143/147) was the most common double mutation and 3 subjects had the double mutation V27I/I203T. A triple mutation, V27I/E114G/I203T, was found in 1 subject. In conclusion, PCR-direct sequencing is a convenient tool for the rapid detection of GJB2 mutations and this data might provide information for the genetic counseling of the GJB2 gene.

• Journal of Food Hygiene and Safety
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• v.24 no.1
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• pp.56-62
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• 2009

Patulin, a mycotoxin mainly produced by Penicillium and Aspergillus, is found in various foods. In the present, a maximum acceptable level for patulin is established at $50{\mu}g/kg(ppb)$ in apple juices and apple concentrates in Korea. But patulin may be detected in foods produced with other fruits. In the present study, patulin contamination was analyzed in 520 samples of fruit juices and beverages, and 50 samples of fruit juice concentrates. High performance liquid chromatography(HPLC) was applied to quantitatively analyze patulin levels in samples and liquid chromatography-mass spectrometry(LC/MS/MS) was used to remove false positive results. The results showed that three samples of 520 fruit juices and beverages and five samples of 50 fruit juice concentrates were contaminated by patulin, $9.8-18.0{\mu}g/kg$ and $4.7-18.2{\mu}g/kg$ respectively. Contaminated samples were produced with apple, orange or pear. This indicates that it is necessary to extend the regulatory range of patulin. In the other hands, the present study confirmed the effectiveness of LC/MS/MS analytical method to remove false positive results.

• Pediatric Infection and Vaccine
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• v.13 no.2
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• pp.106-114
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• 2006

Purpose : Studying the serotypes and emm genotype of group A streptococci(GAS) hold a key role in the investigation of epidemiology, pathogenesis, and resistance to antibiotics. We planed to find out the possible differences in serotypes and genotypes between the invasive and the non-invasive GAS infection. Methods : We obtained 39 isolates from the patients hospitalized in 2004 with the diagnosis of invasive GAS infectious diseases in the Severance Hospital, Yonsei University. We sent the isolates to the WHO Collaboratory Center in University of Minnesota and analyzed T serotypes and emm genotypes. These results were compared with non-invasive GAS infections in our hospital. Results : Compared this data with the results of 2003 to 2004, T2/28, T3, T5/27/44, T9 and NT were more prevalent. The NT showed statistically significant difference(P=0.019). The emm 3, emm 9, emm 18, emm 44 and emm provision type STG485 were more frequent genotypes compared with the study in 2003~2004, relatively. The emm provision type STG485 had statistically significant difference(P=0.000). The incidence of T12 plus T4, known as erythromycin(EM) resistant serotypes, was as low as 17.9% in this study, compared with those of non-invasive strains. Conclusion : The incidence of EM-resistant strains is supposed not to be higher in invasive GAS infection. We could find out possible higher incidence of rheumatogenic strains in the invasive GAS infections. It seems to be important to have an effort on studying of serotyping and genotyping for the monitoring of strains and to know the epidemiologic characteristics.

• Pediatric Infection and Vaccine
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• v.16 no.2
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• pp.175-182
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• 2009

Purpose : A number of countries have experienced an increase in pertussis during the past decade. In particular, there has been an increase in the incidence rate among adolescents and adults. To learn more about the current epidemiology of pertussis, we studied the prevalence and clinical characteristics of pertussis in children in Cheonan, South Korea. Methods : We collected nasopharyngeal aspirates of 118 patients who were treated for respiratory symptoms at Dankook Univeristy Hospital between March 2008 and September 2009. We performed multiplex PCR for detection of Bordetella pertussis in those aspirates. Results : Of the 118 patients, 10 (8%) were positive by PCR for B. pertussis. Six episodes occurred during the period July to September 2009. Nine of the 10 patients were less than 3 months old. Seven of them had not received DTaP vaccine. The mean duration of coughing before diagnosis was 10.9${\pm}$5.2 days. Ten patients (100%) had paroxysmal cough and 8 (80%) had post-tussive vomiting. Only one patient had fever. One who had complications that include pneumonia, atelectasis and pneumomediastinum developed an absolute increase in leukocyte count (84,400/$mm^3$). There was a statistically significant relation between vaccine being received and development of complications (P =0.033). Conclusion : We suspect that there was an epidemic of pertussis between July and September 2009. Further investigation by a pediatric or nationwide surveillance system is needed to monitor the changing epidemiology for pertussis.

• Pediatric Infection and Vaccine
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• v.16 no.2
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• pp.150-161
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• 2009

Purpose : Staphylococcus aureus causes a variety of infections, ranging from benign skin infections to fatal invasive infections. Recently, methicillin-resistant S. aureus (MRSA) infections have emerged in patients who do not have established risk factors. This study was conducted to characterize S. aureus infections in children with an emphasis on communityassociated MRSA infections at a tertiary care pediatric facility during a 3-year period. Methods : Four hundred twenty-nine cases of S. aureus infections diagnosed at the Seoul National University Children's Hospital between January 2004 and December 2006 were retrospectively reviewed. The cases were classified as hospitalonset (HO) or community-onset (CO), healthcare-associated (HA), or community-associated (CA) infections. Results : Among the 206 cases <1 year of age, 72%, 7%, and 21% were HO-HA, CO-HA, and CA infections, respectively, as compared to 48%, 28%, and 24% among the 223 cases >1 year of age. The proportion of CO-HA infections among HA infections (8.6% vs. 37.1%, P<0.001) and the proportion of HA infections among the CO infections (24.5% vs. 54.3%, P <0.001) were greater in older children than in infants. Overall, 57% of the isolates were methicillin-resistant. Twenty-nine (30%) of 96 CA strains were MRSA, and the most common site of CA-MRSA infection was the skin and soft tissues (26 cases). Conclusion : The methicillin resistance rate of S. aureus from CA infections was high and CA-MRSA was most often associated with skin and soft tissue infections.

• Pediatric Infection and Vaccine
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• v.24 no.3
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• pp.134-140
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• 2017

Purpose: Severe combined immunodeficiency (SCID) is the most serious form of primary immunodeficiency. Infants with SCID are susceptible to life-threatening infections. To establish newborn screening for SCID in Korea, we performed a screening test for T-cell receptor excision circle (TREC) and ${\kappa}$-deleting recombination excision circle (KREC) in neonates and investigated the awareness of SCID among their parents. Methods: Collections of dried blood spots from neonates and parent surveys were performed at the Samsung Medical Center and Cheil General Hospital & Women's Healthcare Center in Korea. The amplification crossing point (Cp) value <37.0 was defined as TREC/KRECpositive based on cutoff values from measuring multiplex real-time polymerase chain reaction. A Cp value >39.0 was defined as negative. Results: For TREC/KREC screening, 141 neonates were enrolled; 63 (44.7%) were male. One hundred forty neonates (99.3%) had positive TREC/KREC results at the time of the initial test; 82.3% and 75.9% were positive and 17.0% and 23.4% were weakly positive for TREC and KREC, respectively. In one neonate (0.7%), the initial TREC/KREC test result was negative. However, repeated tests obtained and confirmed a positive result. For an awareness survey, 168 parents were engaged. Only 2% of parents (3/168) knew that the newborn screening test for SCID had been introduced and performed in other countries. Eighty-four percent of parents (141/168) replied that nationwide newborn SCID screening should be performed in Korean newborns. Conclusions: In this study, newborn SCID screening was performed along with assessment of public awareness of the SCID test in Korea. The study results showed that newborn SCID screening can be readily applied for clinical use at a relatively low cost in Korea.

• Pediatric Infection and Vaccine
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• v.25 no.2
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• pp.61-71
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• 2018

Purpose: Research on the clinical role of Staphylococcus aureus as a pathogen in acute gastroenteritis (AGE) in children has been scarce. This study aimed to clarify the prevalence and clinical correlation of S. aureus detection in children with AGE. Methods: Fecal samples were collected from children with symptoms of AGE who visited a secondary hospital between January 2012 and December 2015. The samples were sent to the Seoul Metropolitan Government Research Institute of Public Health and Environment to test for pathogenic organisms. Clinical patterns were analyzed through medical record review. Results: Among the 663 participants, the bacteria detection rate was 26.2% (n=174), the virus detection rate was 29.7% (n=197), and the non-detection rate was 43.1% (n=286). S. aureus was tested positive from 102 cases and was confirmed as a single pathogen in 53 cases. It was the third most common pathogen. The prevalence by age was highest (45.3%) in 0-2 year-olds. Most cases occurred in summer. Symptoms included diarrhea (71.7%), vomiting (67.9%), fever (49.1%), and abdominal pain (37.7%). Only vomiting showed a significant difference between the S. aureus group and the non-detection group (67.9% vs. 43.0%; P=0.001). Among enterotoxins, the higher incidence of vomiting was associated with classical staphylococcal enterotoxins (SEA, SEB, SEC, SED, and SEE) and SEH (P=0.027). Conclusions: S. aureus was the bacteria commonly isolated from children with AGE. Our study identified cases of staphylococcal AGE in children based on fecal samples and confirmed the characteristic symptoms, affected age groups, seasonal distribution, and correlation with enterotoxins.

• Journal of radiological science and technology
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• v.39 no.3
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• pp.421-427
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• 2016

We aim to offer basic materials about infection management through conducting bacterial contamination test about general radiography equipment and CT equipment installed in ER of three general hospitals with 100 sickbeds or more located in Gyeongsangbuk-do Province, and suggest management plan. It had been conducted from 1st December 2015 to 31st December, and objects were general radiography equipment and CT equipment of emergency room located in Gyeongsangbuk-do Province. For general radiography equipment, sources were collected from 4 places such as upper side of control box which employees use most, upper side of exposure button, whole upper side of table which is touching part of patient's skin, upper side of stand bucky's grid, and where patients put their jaws on. For CT equipment, sources were collected from 3 places such as upper side of control box which radiography room employees use most, X-ray exposure button, whole upper side of table which is touching part of patient's skin, and gantry inner. Surface contamination strain found at general radiography equipment in emergency room of radiology are Providencia stuartii(25%), Stenotrophomonas maltophilia(18%), Enterobacter cloacae(8%), Pseudomonas species(8%), Staphylococcus epidermidis(8%), Gram negative bacilli(8%), and ungrown bacteria at incubator after 48 hours of incubation (67%) which is the biggest. Most bacteria were found at upper side of stand bucky-grid and stand bucky of radiology's general radiography equipment, and most sources of CT equipment were focused at patient table, which means it is contaminated by patients who have various diseases, and patients who have strains with decreased immunity may get severe diseases. Thus infection prevention should be made through 70% alcohol disinfection at both before test and after test.