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Development of Multiplex PCR Assay for Identification of Eight Species from Meats in Korea (국내에서 유통되는 8종의 식육감별을 위한 multiplex PCR법 개발)

  • Heo, Eun-Jeong;Ko, Eun-Kyung;Yoon, Hyang-Jin;Kim, Yeon-Hwa;Kim, Young-Jo;Park, Hyun-Jung;Wee, Sung-Hwan;Moon, Jin-San
    • Journal of Food Hygiene and Safety
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    • v.31 no.1
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    • pp.28-35
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    • 2016
  • Species identification of animal tissues in meat products is an important issue to protect the consumer from illegal and/or undesirable adulteration; for economic, religious and health reasons. In this reason, accurate analytical methods are needed for the labeling of meat products with requiring simple and fast procedure. Recently, applications of PCR in food analysis have been increased because of their simplicity, specificity and sensitivity. Therefore, in this study, a multiplex PCR assay was developed for the simultaneous identification of eight species of cow, pig, chicken, duck, goat, sheep, horse and turkey from raw meats. The primers were designed in different regions of mitochondrial 16S RNA after alignment of the available sequences in the GenBank database. Two multiplex primer sets were designed as Set 1 (cow, pig, chicken, duck) and Set 2 (goat, sheep, horse, turkey), respectively. Total 274 samples from cow (n = 55), pig (n = 30), chicken (n=30), and duck (n = 30), goat (n = 40), sheep (n = 33), horse (n = 41), and turkey (n = 15) were tested. The primers generated specific fragments of 94, 192, 279, 477 bp (pig, chicken, cow, duck), 670, 271, 152, 469 bp (goat, sheep, horse, turkey) lengths for eight species, respectively. The animal species specificity was 100% in all eight samples in the multiplex PCR assay. The detection limit of the multiplex PCR assay showed from 100 fg to 1 pg of template DNA from extracted from raw meats. When applying multiplex PCR assays to sample from pork/beef and pork/chicken, beef/chicken tested raw mixed meats and heat-treated ($83^{\circ}C$ for 30min, $100^{\circ}C$ for 20min, and $121^{\circ}C$ for 10min) mixtures, detection limit was 0.1% level beef, pork and pork in beef and chicken in pork and 1.0% level pork in chicken. This study suggest that the developed multiplex PCR assay can be used for rapid and simultaneous species identification of cow, pig, chicken, duck, goat, sheep, horse and turkey from meats.

Survey on Comparative Awareness between Media and Industry on Occurrence of Food Safety Incident (식품안전사건 발생 시 미디어와 산업의 인식도 조사)

  • Shin, Won-Jung;Lee, Yu-Si;Oh, Se-Ra;Park, Tae-Gyun;Kim, Renee;Kim, Ho-Sik;Lee, Jung-Ho;Lee, Seung-Yong;Ha, Sang-Do
    • Journal of Food Hygiene and Safety
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    • v.28 no.2
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    • pp.108-114
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    • 2013
  • To describe the comparative awareness between media and industry, we conducted a survey with a professional group including 28 food safety managers at food corporations and 17 food journalists to find out the differences in perception of 'reporting terms' and 'correspondence terms' when food safety incident occurs. We surveyed by questionnaire methods and did the statistical analysis by SPSS. The results showed that 70.6% of the journalists and 92.9% of the corporation workers answered 'yes indeed' about the question whether the media has a great influence when food safety incident occurs. In the perception that use of the stimulating terms of the media was the cause of company damage, 83% of the corporate workers answered 'yes indeed', and 70.6% of the journalists answered 'yes'. It means that both groups agreed on stimulating terms causing company damage. In the question whether the 'reporting terms' of the media and the 'correspondence terms' of the government and academia made a difference in the consumers' perception and the spread of the incident, 92.9% of the corporate workers and 35.3% of the journalists answered 'yes indeed'. It means that both groups showed a common opinion that the difference in choice of words expanded the perception of consumers, and the spread of the incident. However, the corporate workers agreed with this in a more serious manner than the journalist group. In standardization of 'reporting terms' and restricting use, corporate workers were overall at a positive stand while journalists showed a big difference in opinion within the group. In conclusion, it is necessary to establish a measure to standardize the terms used by corporations, journalists, and professionals in order to decrease the differences of terms and the damage of consumers and the company when food safety incident occurs.

Effect of Frying Methods under Reduced Pressures on the Oxidative Stability of Frying Oils (감압 튀김 방법이 산패 변화에 미치는 영향)

  • Lee, Bo-Bae;Lee, Jin-Won;Park, Jang-Woo;Chung, Yoon-Kyung;Lee, Hyun-Joo
    • The Korean Journal of Food And Nutrition
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    • v.26 no.1
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    • pp.15-21
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    • 2013
  • This study investigated the oxidative stability of oils when dough was fried under a lower pressure than the ambient atmosphere. The pressure during the frying process was controlled at measures of 760, 560, 360 or 160 mmHg. The oil containing the dough was heated at $180^{\circ}C$ for 48 hours. Rancidity values, including acid value, peroxide value, fatty acid analysis, color changes, and browning of oil samples, were measured every 8 hours. As the frying process continued at all 4 pressure levels, the acid values (AV) increased. However, compared to the other pressure levels, the increase in AV was the least at 160 mmHg. In addition, the peroxide value at 160 mmHg was only 0.81 meq/kg compared to 1.52 meq/kg at 760 mmHg. For all pressure levels, stearic acid, oleic acid, ${\omega}$-6 linolenic acid were increased, while linoleic acid and ${\omega}$-3 linolenic acid were decreased. In terms of color, a-values representing redness were decreased, whereas b-values were increased as the frying proceeded. These results revealed that the oxidation of frying oil was decreased under reduced pressure condition. Thus, the usage of frying oil may be extended, owing to less oxidative concerns. This leads to a lower cost to the manufacturer, and furthermore, helps the environment by reducing industrial wastes.

Influence of Environmental Living Standards on Helicobacter pylori Infection in Korean Elementary School Children (서울 지역 초등학생의 생활환경과 Helicobacter pylori 양성률)

  • Kim, Je-Woo;Kim, Hyo-Shin;Chung, Ki-Sup
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.4 no.1
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    • pp.10-17
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    • 2001
  • Purpose: We measured anti-H. pylori IgG in Korean elementary school children living in Shinchon area of Seoul, Korea to evaluate the influence of environmental living standards on H. pylori infection. Methods: IgG antibodies to H. pylori were measured in plasma using a commercial ELISA kit (GAP IgG Helicobacter pylori, Bio-Rad Laboratories Inc., Hercules, CA, USA). Information on environmental status such as place of birth, parental income, type of housing, number of persons in the household, parents' occupation, family history of peptic ulcer disease and gastric cancer was obtained. Statistical analysis was done by Chi-square and logistic regression test using SPSS $7.0^{TM}$ for Windows. Results: Study subjects consisted of 571 children, and the age distribution ranged from 6.0 to 13.6 years with a mean of $9.6{\pm}1.8$ years. Male-to-female ratio was 1.1:1. The seropositive rates of H. pylori infection ranged from 10.4% in children aged 6 years to 30.9% in 12 year-old group, overall 16.8%. The prevalence of H. pylori infection progressively increased with age, but there was no significant difference in seropositive rates among children in different age groups (p=0.06). Seropositive rates of anti-H. pylori IgG on the basis of gender, place of birth, parental income, type of housing, parents' occupation, family history of peptic ulcer disease and gastric cancer showed no statistically significant difference. Interestingly, however, seropositive rate of anti-H. pylori IgG showed statistical significance in relation to number of persons in the household (p=0.003; Odds ratio 1.50 by logistic regression test). Conclusion: These results suggest that number of persons in the household is the most important factor among environmental living standards, and that risk of H. pylori infection increases by increment of 1.5 times as the number of persons in the household increases by one.

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Analysis of the Tumor Necrosis Factor-${\alpha}$ Promoter Polymorphism in Children with Henoch-Sch$\"{o}$nlein Purpura (Henoch-Sch$\"{o}$nlein 자반증에서 Tumor Necrosis Factor-${\alpha}$ 유전자 다형성 분석)

  • Yang, Hye-Ran;Ko, Jae-Sung;Seo, Jeong-Kee
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.10 no.1
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    • pp.11-19
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    • 2007
  • Purpose: Henoch-Sch$\"{o}$nlein purpura (HSP) is a systemic vasculitis involving the skin, joints, gastrointestinal tract, and kidney. Although the pathogenesis of HSP is still unclear, tumor necrosis factor (TNF-${\alpha}$) is regarded as an important cytokine contributing to the disease. The goal of this study was to determine the role of TNF-${\alpha}$ in the pathogenesis of HSP, and to evaluate the TNF-${\alpha}$ polymorphism for genetic susceptibility to HSP. Methods: From March 2004 to November 2005, 40 children with HSP and 32 healthy controls were included. Serum TNF-${\alpha}$ levels were measured using the ELISA method during the acute and convalescent phase of HSP. The genotypic and allelic frequencies of the TNF-${\alpha}$ gene polymorphisms at positions -308 and -238 were evaluated in patients and controls. Results: Serum TNF-${\alpha}$ levels were $23.17{\pm}11.31$ pg/mL in the acute phase of children with HSP and $10.56{\pm}5.59$ pg/mL in the convalescent phase (p=0.000). There was no significant correlation between the serum TNF-${\alpha}$ levels and the clinical scores of HSP (r=0.310, p=0.070). The genotypic frequency of the TNF-${\alpha}$ -308 polymorphism in children with HSP was not significantly different compared to healthy controls (GG 80%, GA 20% vs. GG 93.8%, GA 6.2%; p=0.094). The genotypic frequency of the TNF-${\alpha}$ -238 polymorphism in children with HSP was not significantly different (GG 97.5%, GA 2.5% vs. GG 93.8%, GA 6.3%; p=0.429). Conclusion: TNF-${\alpha}$ is assumed to be the main cytokine associated with the pathogenesis of HSP during the acute phase. However, the presence of TNF-${\alpha}$ gene polymorphisms at positions -308 and -238 did not distinguish children with HSP from normal controls.

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Analysis of 1,000 Cases of Abdominal Ultrasonography Performed by a Pediatrician (소아과 의사에 의해 시행된 복부 초음파 검사 1,000예에 대한 분석)

  • Bae, Sang-In;Park, Jae-Hong
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.10 no.1
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    • pp.28-35
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    • 2007
  • Purpose: The aim of this study was to evaluate the clinical usefulness of ultrasound examination of children performed by a pediatrician. Methods: One thousand children who presented with symptoms of a gastrointestinal disorder and underwent abdominal ultrasound evaluation in the Department of Pediatrics, between January 2003 and June 2006, were included in this study. We analyzed the patient's medical records and ultrasound results retrospectively. Results: Among the 1,000 patients, 58.4% were male and 41.6% were female. The mean age of the patients was $4.7{\pm}4.0$ years. The main reasons for ultrasound were abdominal pain (43.9%), vomiting (17.3%), elevated liver enzymes (11.8%), and jaundice (9.8%). Abnormal ultrasound findings were present in 57.9% of cases. The major abnormal findings were mesenteric lymphadenitis (29.2%), fatty liver (12.1%), hepatitis (6.4%), hepatosplenomegaly (6.2%), and acute appendicitis (4.8%). The time interval between the initial medical evaluation and the ultrasound evaluation was within 24 hours in most cases (78.5%). The main findings in children with abdominal pain were mesenteric lymphadenitis (32.6%), fatty liver (5.9%), intussusception (2.7%), and acute appendicitis (2.7%). The main findings in children with vomiting were mesenteric lymphadenitis (12.7%), hypertrophic pyloric stenosis (10.4%), and acute appendicitis (3.5%). The major ultrasound findings in children with urinary tract diseases were hydronephrosis (45.4%), urolithiasis (21.5%) and cystic renal disease (18.1%). Conclusion: Ultrasound examination played an important role as a non-invasive and prompt screening examination for detection of abdominal diseases. Ultrasound was an important tool for pediatricians to determine timely information for patient management.

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Adenoviral Lower Respiratory Tract Infections in Children; Serotypes and Clinical Characteristics (소아의 아데노바이러스 하기도 감염증: 혈청형 및 임상상)

  • Shin, Seon Hee;Lee, Hoan Jong;Yoon, Hae Sun
    • Pediatric Infection and Vaccine
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    • v.9 no.2
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    • pp.182-192
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    • 2002
  • Purpose : This study was performed to characterize the epidemiologic and clinical features of acute adenoviral lower respiratory tract infections(LRTIs). Methods : Virological analysis was done from respiratory specimens obtained from patients with LRTIs hospitalized to other hospitals and referred to the Department of Pediatrics, Seoul National University Children's Hospital(SNUCH) from June 1998 to July 2000. Viral diagnosis was made by isolation of viruses employing HEp-2 cell culture and indirect immunofluorescent staining with monoclonal antibodies. Serotypes of adenoviruses were determined by neutralization test using antiserum for types 1, 2, 3, 4, 5, 6, 7 and 11. Medical records of children admitted to the SNUCH were reviewed retrospectively. Results : Adenovirus was isolated from 118(9.0%) of 1,305 children with LRTIs. Serotypes were 3(39.0%), 7(16.9%), 1(11.0%), 2(7.6%), 4(7.6%), 6(5.9%), 11(2.5%), and 5(0.8%) and 10 strains(8.5%) were not neutralized by antisera included in the study. Infections by type 3 and type 7 occurred in outbreaks. Male to female ratio was 1.0:0.9 and mean age was 1.95 years. The clinical diagnoses were pneumonia(83%), acute tracheobronchitis(12%) and bronchiolitis(5%). Associated symptoms, signs and abnormal laboratory findings included cough(100%), sputum(73.5%), fever(54.2%), rale(59.3%), wheezing(34%), anemia(35%) and leukopenia(15.8%). Mortality was in 13.5%. Residual radiologic sequelae was identified in 32.6% of the patients followed. Conclusion : These data confirms that adenovirus may cause severe lower respiratory tract diseases, and infections by type 3 and 7 may occured in outbreaks.

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Diagnostic Value of Serum Procalcitonin in Febrile Infants Under 6 Months of Age for the Detection of Bacterial Infections (발열이 있는 6개월 미만의 영아에서 세균성 감염에 대한 procalcitonin의 진단적 가치)

  • Kim, Nam Hyo;Kim, Ji Hee;Lee, Taek Jin
    • Pediatric Infection and Vaccine
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    • v.16 no.2
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    • pp.142-149
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    • 2009
  • Purpose : The aim of this study was to determine the diagnostic value of serum procalcitonin (PCT) compared with that of C-reactive protein (CRP) and the total white blood cell count (WBC) in predicting bacterial infections in febrile infants<6 months of age. Methods : A prospective study was performed with infants <6 months of age who were admitted to the Department of Pediatrics with a fever of uncertain source between July and September 2008. Spinal taps were performed according to clinical symptoms and physical examination. Serum PCT levels were measured using an enzyme-linked fluorescent assay. Results : Seventy-one infants (mean age, 2.62 months) were studied. Twenty-six infants (36.6%) had urinary tract infections (UTIs), and 22 infants (31.0%) had viral meningitis. The remaining infants had acute pharyngitis (n=1), herpangina (n=1), upper respiratory tract infections (n=7), acute bronchiolitis (n=8), acute gastroenteritis (n=4), and bacteremia (n=2). The median WBC and CRP levels were significantly higher in infants with UTIs than in infants with viral meningitis. However, there were no differences in the median PCT levels between the groups (0.14 ng/mL vs. 0.11 ng/mL, P=0.419). The area under the receiver operating characteristic curve was 0.792 (95% CI, 0.65-0.896) for WBC, 0.77 (95% CI, 0.626-0.879) for CRP, and 0.568 (95% CI, 0.417-0.710) for PCT. An elevated WBC count (>11,920/${\mu}L$) and an increased CRP level (>1.06mg/dL) were significant predictors of UTIs based on multiple logistic regression analysis. Conclusion : Serum PCT concentrations should be interpreted with caution in infants <6 months of age with a fever of uncertain source.

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A Retrospective Analysis of Characteristics of Probiotics Associated with Invasive Bacterial Infections in Children (소아청소년에서 발생한 유산균 제제에 포함된 균주에 의한 침습성 감염증의 후향적 분석)

  • Koh, June Young;Seo, Euri;Lee, Jina
    • Pediatric Infection and Vaccine
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    • v.24 no.3
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    • pp.168-177
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    • 2017
  • Purpose: The purpose of this study was to analyze the clinical features and risk factors of invasive infections caused by Lactobacillus spp. and Saccharomyces spp., components of commercially available probiotics. Methods: We analyzed demographic and clinical data from children ${\leq}18$ years of age with an invasive infection caused by Lactobacillus spp. or Saccharomyces spp. at the Asan Medical Center Children's Hospital from January 1998 to June 2016. Probiotic consumption data were also analyzed. Results: During the study period, a total of 24 episodes of invasive infections were caused by Lactobacillus spp. (n=16) and Saccharomyces cerevisiae (n=8). Along with the increase of probiotic use (755,594 [days/1,000 patient-admission days] in 2001 to 2005, 1,444,066 in 2006 to 2010, and 6,904,736 in 2011 to 2016), the incidence of probiotic-associated invasive infection increased ($R^2=0.70$). The median age of the patients was 1.8 years (range, 2 months to 17 years), and most of them had underlying medical conditions. The 30-day mortality rate was 20.8% (5/24), and 11 (45.8%) of these patients resulted from a severe invasive infection. We determined the risk factors for invasive infection to be: previous intensive care unit stay (odds ratio [OR], 3.0; 95% confidence interval [CI], 1.5 to 6.1] and the presence of a central venous catheter (OR, 2.2; 95% CI, 1.2 to 4.3). Conclusions: Although the probiotic-associated invasive infections rarely occurred in children, the incidence has increased along with probiotic pressure. Judicious use of probiotics is mandatory, especially in young children with underlying medical conditions and continuous surveillance will be needed to minimize the safety concerns.

Evaluation of Antibodies Against Haemophilus influenzae Type b in Korean Adults (우리나라 성인에서 Haemophilus influenzae type b에 대한 항체 평가)

  • Lee, Ji Hyen;Kim, Han Wool;Lee, Soyoung;Kim, Kyung-Hyo
    • Pediatric Infection and Vaccine
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    • v.24 no.3
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    • pp.125-133
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    • 2017
  • Purpose: After the introduction of Haemophilus influenzae type b (Hib) vaccine in 1995 in Korea, it was included in the national immunization program in 2013. In the post-Hib vaccine era, some studies in other countries reported that invasive Hib disease affects adults, especially the elderly and immunocompromised persons, more often than it affects children. To evaluate disease susceptibility, quantitative and qualitative analysis of anti-polyribosylribitol phosphate (PRP) antibodies were carried out in Korean adults aged 20 to 85 years. Methods: Sera were collected from 39 healthy adults (20 to 50 years of age) and from 30 elderly adults (75 to 85 years of age) who did not have immune-compromising conditions. The concentration of anti-PRP immunoglobulin G (IgG) and serum bactericidal indices (SBIs) were measured by enzyme-linked immunosorbent assay and serum bactericidal assay. Results: Geometric mean concentrations of anti-PRP IgG and geometric mean SBIs were $0.88{\mu}g/mL$ (95% confidence interval [CI], 0.17 to 3.85) and 354 (95% CI, 50 to 2,499) in young adults and $1.67{\mu}g/mL$ (95% CI, 0.53 to 5.24) and 449 (95% CI, 146 to 1,376) in elderly adults, respectively. When the threshold of seropositivity for anti-PRP IgG was applied as 0.15 or $1.0{\mu}g/mL$, which is the protective antibody level in children, seropositive rates were 87.2% or 53.8% in young adults and 100% or 60% in elderly adults. The seropositivity rates of the SBI ($SBI{\geq}4$) were 82.1% and 100% in the groups, respectively. Conclusions: Most subjects in the adult and elderly adult groups display immunity to Hib based on quantitative and qualitative antibody levels, but not all. Because high immunization and low Hib circulation rates may reduce the natural Hib immunity in the population, monitoring Hib immunity as well as disease are needed continuously.