• Title/Summary/Keyword: Multiplex culture

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Biological Hazard Analysis of Paprikas, Strawberries and Tomatoes in the Markets (유통중인 파프리카, 딸기 및 토마토의 생물학적 위해요소 분포 조사)

  • Yu, Yong-Man;Youn, Young-Nam;Hua, Quan Juan;Cha, Guang-Ho;Lee, Young-Ha
    • Journal of Food Hygiene and Safety
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    • v.24 no.2
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    • pp.174-181
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    • 2009
  • The consumption of "ready-to-eat" agricultural products is recently increasing and the safety of these agricultural products is forefront of public concerns. The 120 samples of paprikas, strawberries and tomatoes, which are the representative exported agricultural products, were purchased at the department stores and discount stores in Daejeon. And we determined the microbiological and parasitological contamination level of these agricultural products using culture media, multiplex PCR, commercial bacterial detection kit and microscopy, and also evaluated the decontamination method. Mean counts of total aerobic bacteria from these agricultural products ranged from $1.3{\times}10^4$ CFU/g to $1.8{\times}10^5$ CFU/g, and mean counts of coliforms ranged from $1.4{\times}10^3$ CFU/g to $9.6{\times}10^3$ CFU/g. There was no significant difference in the level of bacterial contamination between the agricultural products from department stores and the ones from discount stores. Strawberry showed the highest contamination level for the bacteria and we also found the unidentified parasite eggs. Enterobacter cloacae was the most frequently isolated bacteria strain, but no food poisoning pathogenic bacteria except Staphylococcus aureus was isolated from the products by multiplex PCR. Compared to unwashed products, tab water-washed ones showed 80% decrease of the counts of total aerobic bacteria on the agricultural products, and the rates decreased more by incorporating detergent or ultrasonic wave treatment. We concluded that the biological contamination levels among paprikas, strawberries and tomatoes were the highest in strawberries, but there were not significant difference according to distribution systems.

Diagnostic value of the Vesikari Scoring Sys­tem for predicting the viral or bacterial patho­gens in pediatric gastroenteritis

  • Shim, Dong Ho;Kim, Dong Yeon;Cho, Ky Young
    • Clinical and Experimental Pediatrics
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    • v.59 no.3
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    • pp.126-131
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    • 2016
  • Purpose: To evaluate the diagnostic value of the Vesikari Scoring System (VSS) as an early predictor of pathogens in children with acute gastroenteritis (AG). Methods: In this retrospective study, the VSS score, absolute neutrophil count (ANC), and C-reactive protein (CRP) levels were analyzed in 107 hospitalized children with AG, aged 6 months to 17 years. Patients were divided into nonspecific, viral, and bacterial groups according to the pathogens detected using a multiplex polymerase chain reaction (PCR) test. Results: Patients in the bacterial group had significantly higher CRP values and VSS scores compared to those in the viral group and significantly higher VSS scores compared to those in the nonspecific group (P<0.05). Patients in the viral group had significantly higher VSS scores than those in the nonspecific group (P<0.05). Logistic regression analysis revealed that VSS was the most effective diagnostic tool for predicting the type of pathogen (P<0.05). The area under the receiver operating characteristics curve of VSS was significantly greater than that for ANC and CRP (P<0.05). At a cutoff point of 10 in the VSS, an acceptable diagnostic accuracy could be achieved for distinguishing between bacterial and viral pathogens in AG. Conclusion: VSS can be considered a useful and reliable infectious marker for pediatric gastroenteritis. VSS may be a good early predictor of the type of pathogen, enabling development of a treatment plan before results from a stool culture or PCR test are available.

Inhibitory Effect of Gallic acid on Production of Interleukins in Mouse Macrophage Stimulated by Lipopolysaccharide (Gallic acid가 Lipopolysaccharide로 활성화된 마우스 대식세포의 인터루킨 생성에 미치는 영향)

  • Park, Wan-Su
    • Journal of Pharmacopuncture
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    • v.13 no.3
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    • pp.63-71
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    • 2010
  • Objectives: Gallic acid (GA) is the major component of tannin which could be easily founded in various natural materials such as green tea, red tea, grape juice, and Corni Fructus. The purpose of this study is to investigate the effect of Gallic acid (GA) on production of interleukin (IL) in mouse macrophage Raw 264.7 cells stimulated by lipopolysaccharide (LPS). Methods: Productions of interleukins were measured by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$ (multi-analyte profiling beads) technology. Firstly, cell culture supernatant was obtained after treatment with LPS and GA for 24 hour. Then, it was incubated with the antibody-conjugated beads for 30 minutes. And detection antibody was added and incubated for 30 minutes. And Strepavidin-conjugated Phycoerythrin (SAPE) was added. After incubation for 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System and concentration of interleukin was determined. Results: The results of the experiment are as follows. 1. GA significantly inhibited the production of IL-3, IL-10, IL-12p40, and IL-17 in LPS-induced mouse macrophage RAW 264.7 cells at the concentration of 25, 50, 100, 200 uM (p<0.05). 2. GA significantly inhibited the production of IL-6 in LPS-induced mouse macrophage RAW 264.7 cells at the concentration of 50, 100, 200 uM (p<0.05). 3. GA diminished the production of some cytokine such as IL-4, IL-5, and IL-13 in LPS-induced mouse macrophage RAW 264.7 cells. 4. GA did not show the inhibitory effect on the production of IL-$1{\alpha}$ and IL-9 in LPS-induced mouse macrophage RAW 264.7 cells. Conclusions: These results suggest that GA has anti-inflammatory activity related with its inhibitory effects on the production of interleukins such as IL-3, IL-10, IL-12p40, IL-17, and IL-6 in LPS-induced macrophages.

Detection of Foodborne Pathogens and Mycotoxins in Eggs and Chicken Feeds from Farms to Retail Markets

  • Lee, Minhwa;Seo, Dong Joo;Jeon, Su Been;Ok, Hyun Ee;Jung, Hyelee;Choi, Changsun;Chun, Hyang Sook
    • Food Science of Animal Resources
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    • v.36 no.4
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    • pp.463-468
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    • 2016
  • Contamination by foodborne pathogens and mycotoxins was examined in 475 eggs and 20 feed samples collected from three egg layer farms, three egg-processing units, and five retail markets in Korea. Microbial contamination with Salmonella species, Escherichia coli, and Arcobacter species was examined by bacterial culture and multiplex polymerase chain reaction (PCR). The contamination levels of aflatoxins, ochratoxins, and zearalenone in eggs and chicken feeds were simultaneously analyzed with high-performance liquid chromatography coupled with fluorescence detection after the post-derivatization. While E. coli was isolated from 9.1% of eggs, Salmonella species were not isolated. Arcobacter species were detected in 0.8% of eggs collected from egg layers by PCR only. While aflatoxins, ochratoxins, and zearalenone were found in 100%, 100%, and 85% of chicken feeds, their contamination levels were below the maximum acceptable levels (1.86, 2.24, and 147.53 μg/kg, respectively). However, no eggs were contaminated with aflatoxins, ochratoxins, or zearalenone. Therefore, the risk of contamination by mycotoxins and microbes in eggs and chicken feeds is considered negligible and unlikely to pose a threat to human health.

Inhibitory Effect of Gallic acid on Production of Chemokine and Growth Factor in Mouse Macrophage Stimulated by Lipopolysaccharide (Gallic acid가 Lipopolysaccharide로 활성화된 마우스 대식세포의 케모카인과 성장인자 생성에 미치는 영향)

  • Park, Wan-Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.24 no.4
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    • pp.586-591
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    • 2010
  • Chemokine and Growth Factor are major mediumtors of immuno-inflammatory pathway. The purpose of this study is to investigate whether productions of Chemokine and Growth Factor in lipopolysaccharide (LPS)-induced mouse macrophage RAW 264.7 cells are modulated by Gallic acid (GA), which is easily founded in tannin-containing natural materials such as red wine, green tea, grape juice, and Corni Fructus. Productions of Chemokine and Growth Factor were analyzed by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$ (multi-analyte profiling beads) technology. At first, cell culture supernatant was obtained after treatment with LPS and GA for 24 hour. Then, the antibody-conjugated beads were added and incubated for 30 minutes. After incubation, detection antibody was added and incubated for 30 minutes. And Strepavidin-conjugated Phycoerythrin (SAPE) was added. After incubation for 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System. Based on fluorescence intensity, concentrations of Chemokine and Growth Factor were determined. The results of the experiment are as follows. GA significantly inhibited the production of interferon-inducible protein (IP)-10, keratinocyte-derived chemokine(KC), and vascular endothelial growth factor (VEGF) in LPS-induced RAW 264.7 cells at the concentration of 25, 50, 100, 200 uM (p<0.05). GA significantly inhibited the production of monocyte chemoattractant protein-1(MCP-1) and macrophage-colony stimulating factor(M-CSF) in LPS-induced RAW 264.7 cells at the concentration of 50, 100, 200 uM (p<0.05). GA diminished the production of granulocyte macrophage-colony stimulating factor (GM-CSF) in LPS-induced RAW 264.7 cells. But GA did not show the inhibitory effect on the production of leukemia inhibitory factor (LIP) and macrophage inflammatory protein (MIP)-2 in LPS-induced RAW 264.7 cells. These results suggest that GA has the immuno-modulating activity related with its inhibitory effects on the production of IP-10, KC, MCP-1, VEGF, and M-CSF in LPS-induced macrophages.

The Effect of Bacillus-Fermented Scutellariae Radix Acupuncture Solution on Chemokine and Growth Factor Production in Mouse Macrophage Stimulated by Lipopolysaccharide (바실러스균 발효황금약침액이 마우스 대식세포의 케모카인 및 생성인자 생성에 미치는 영향)

  • Park, Wan-Su
    • Korean Journal of Acupuncture
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    • v.27 no.3
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    • pp.109-118
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    • 2010
  • Objectives : The purpose of this study is to investigate the effect of Bacillus-fermented Scutellariae Radix acupuncture solution (SB) on chemokine and growth factor production in RAW 264.7 mouse macrophages stimulated by lipopolysaccharide (LPS). Methods : Productions of chemokine and growth factor were measured by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on xMAP$^{(R)}$ technology. Firstly, cell culture supernatant was obtained after treatment with LPS (1 ${\mu}g$/mL) and SB for 24 hours. Then, it was incubated with the antibody-conjugated beads for 30 minutes. Detection antibody was then added and incubated for 30 minutes. After incubated for 30 minutes, strepavidin-conjugated phycoerythrin (SAPE) was added. After another 30 minutes incubation, the level of SAPE fluorescence was analyzed in Bio-plex Suspension Array System. Results : The results of the experiment are as follows. 1. SB significantly inhibited the LPS-induced production of vascular endothelial growth factor (VEGF), interferon-inducible protein (IP)-10, and granulocyte-colony stimulating factor (G-CSF) at the concentration of 25, 50, 100, and 200 ${\mu}g$/mL in RAW 264.7 cells (P < 0.05). 2. SB significantly inhibited the LPS-induced production of Eotaxin at the concentration of 25, 100, and 200 ${\mu}g$/mL in RAW 264.7 cells (P < 0.05). 3. SB significantly inhibited the LPS-induced production of MIP-$1\alpha$ at the concentration of 25 and 100 ${\mu}g$/mL in RAW 264.7 cells (P < 0.05). Conclusions : These results suggest that SB has immuno-modulatory property related with its inhibition of VEGF, IP-10, G-CSF, and Eotaxin production in macrophages.

Investigation of Flavor-Forming Starter Lactococcus lactis subsp. lactis LDTM6802 and Lactococcus lactis subsp. cremoris LDTM6803 in Miniature Gouda-Type Cheeses

  • Lee, Hye Won;Kim, In Seon;Kil, Bum Ju;Seo, Eunsol;Park, Hyunjoon;Ham, Jun-Sang;Choi, Yun-Jaie;Huh, Chul Sung
    • Journal of Microbiology and Biotechnology
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    • v.30 no.9
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    • pp.1404-1411
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    • 2020
  • Lactic acid bacteria (LAB) play an important role in dairy fermentations, notably as cheese starter cultures. During the cheese production and ripening period, various enzymes from milk, rennet, starter cultures, and non-starter LABs are involved in flavor formation pathways, including glycolysis, proteolysis, and lipolysis. Among these three pathways, starter LABs are particularly related to amino acid degradation, presumably as the origins of major flavor compounds. Therefore, we used several enzymes as major criteria for the selection of starter bacteria with flavor-forming ability. Lactococcus lactis subsp. lactis LDTM6802 and Lactococcus lactis subsp. cremoris LDTM6803, isolated from Korean raw milk and cucumber kimchi, were confirmed by using multiplex PCR and characterized as starter bacteria. The combinations of starter bacteria were validated in a miniature Gouda-type cheese model. The flavor compounds of the tested miniature cheeses were analyzed and profiled by using an electronic nose. Compared to commercial industrial cheese starters, selected starter bacteria showed lower pH, and more variety in their flavor profile. These results demonstrated that LDTM6802 and LDTM6803 as starter bacteria have potent starter properties with a characteristic flavor-forming ability in cheese.

Clinical Significance of Inflammatory Biomarkers in Acute Pediatric Diarrhea

  • Park, Yoonseon;Son, Minji;Jekarl, Dong Wook;Choi, Hyun Yoo;Kim, Sang Yong;Lee, Seungok
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.22 no.4
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    • pp.369-376
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    • 2019
  • Purpose: The aim of this study was to evaluate the clinical significance of inflammatory biomarkers in acute infectious diarrhea among children. Methods: Clinical parameters including fever, bacterial and viral etiology based on stool culture and multiplex polymerase chain reaction, and nine biomarkers including C-reactive protein (CRP), erythrocyte sedimentation rate (ESR) and leukocytes in blood and calprotectin, lactoferrin, myeloperoxidase, polymorphonuclear elastase, leukocytes, and occult blood in feces were evaluated in children who were hospitalized due to acute diarrhea without underlying disease. Results: A total of 62 patients were included. Among these patients, 33 had fever, 18 showed bacterial infections, and 40 patients were infected with 43 viruses. Of all the biomarkers, CRP was significantly correlated with fever (p<0.001). CRP, ESR, calprotectin, lactoferrin, myeloperoxidase, fecal leukocytes, and occult blood were significantly associated with infection with bacterial pathogens (p<0.001, p=0.04, p=0.03, p=0.003, p=0.02, p=0.03, p=0.002, respectively). The combination of CRP and fecal lactoferrin at their best cut-off values (13.7 mg/L and $22.8{\mu}g/mL$, respectively) yielded a sensitivity of 72.2%, and a specificity of 95.5% for bacterial etiology compared with their individual use. Conclusion: Blood CRP is a useful diagnostic marker for both fever and bacterial etiology in acute pediatric diarrhea. The combination of CRP and fecal lactoferrin yields better diagnostic capability for bacterial etiology than their use alone for acute diarrhea in children without underlying gastrointestinal disease.

The study of caries activity test by multiplex-quantity real time PCR with easy perio test (구강세균 유전자 검사(easy perio test)법을 이용한 치아우식 검사)

  • Yun, Han Gyeol;Park, Seong Gyu;Kim, Jin
    • Journal of Korean Academy of Dental Administration
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    • v.6 no.1
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    • pp.11-18
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    • 2018
  • The aim of this study was to evaluate the competency of the Easyperio test, a genetic test method based on real time PCR for the detection of bacteria that cause dental caries and periodontal disease. To verify the validity of this text, various dental health evaluations were administered to 33 boys between the ages of 12 to 14, as this age group commonly experiences dental caries. These evaluations included a dental caries experience survey, a first molar health evaluation, the Dentocult Streptococcus mutans (SM) strip mutans, the Dentocult Lactobacillus spp (LB) test, and the Easyperio test. The correlation coefficients between the level of the Dentocult SM strip mutans and the dental caries experience were DT (R=0.570, p=0.001), DMFT (R=0.376, p=0.031), and first molar health (R=-0.395, p=0.023). The correlation coefficients between the amount of SM in the Easyperio test and dental caries experience were DT (R=0.528, p=0.002), DMFT (R=0.369, p=0.035), and first molar health (R=-0.426, p=0.013). The correlation coefficients between the level of the dentocult SM strip mutans and the SM amounts of the Easyperio test were S.mi (R=0.564 p=0.001) and S.mu (R=0.621, p=0.002). The correlation coefficients between the level of the Dentocult LB test and the SM amount of Easyperio test was S.mi (R=0.495, p=0.003). In conclusion, Easyperio test may be an easy and effective method for the differentiation and diagnosis of dental caries through quantitative and qualitative analysis of oral bacteria.

Campylobacter Enteritis: Clinical Features and Laboratory Findings in Children Treated at a Single Hospital

  • Jang, Won Tae;Jo, Na Hyun;Song, Mi Ok;Eun, Byung Wook;Ahn, Young Min
    • Pediatric Infection and Vaccine
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    • v.26 no.1
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    • pp.22-31
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    • 2019
  • Purpose: Campylobacter species are common causes of bacterial enteritis. There is limited knowledge on its prevalence and clinical features because of its fastidious culture conditions. The purpose of this study was to identify the clinical features of Campylobacter enteritis in children. Methods: We obtained stool specimens from patients diagnosed with acute gastroenteritis in the Department of Pediatrics, Nowon Eulji Medical Center (NEMC) and identified the pathogens by performing cultures or polymerase chain reactions (PCR). We retrospectively reviewed the medical records of patients with Campylobacter enteritis at NEMC between January 2012 and December 2017. Results: Overall, 123 patients were diagnosed with Campylobacter enteritis (60 by culture and PCR in EnterNet and 110 by multiplex PCR). The median (interquartile range [IQR]) age of patients was 12 years (IQR, 8 to 16 years). The disease occurred all year round, but 69.9% from June to September. Symptoms included diarrhea (97.6%), fever (96.7%), abdominal pain (94.3%), vomiting (37.4%), and headache (34.1%). Compared with other treatments, treatment with azithromycin was associated with a shorter hospitalization period (P<0.05). Conclusions: Campylobacter enteritis is common during summer and mostly infects adolescent patients. It causes severe abdominal pain and fever preceding the onset of diarrhea. Prompt diagnosis and appropriate use of antibiotics reduces the duration of the disease.