• 제목/요약/키워드: Pathogen detection

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Diagnosis of Enteropathogens in Children with Acute Gastroenteritis: One Year Prospective Study in a Single Hospital (소아의 급성 위장관염의 원인균 진단: 단일 병원에서 1년간의 전향적 연구)

  • Chang, Ju Young;Choi, Ji Eun;Shin, Sue;Yoon, Jong Hyun
    • 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.

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Quantitative PCR for Etiologic Diagnosis of Methicillin-Resistant Staphylococcus aureus Pneumonia in Intensive Care Unit

  • Kwon, Sun-Jung;Jeon, Tae-Hyeon;Seo, Dong-Wook;Na, Moon-Joon;Choi, Eu-Gene;Son, Ji-Woong;Yoo, Eun-Hyung;Park, Chang-Gyo;Lee, Hoi-Young;Kim, Ju-Ock;Kim, Sun-Young;Kang, Jae-Ku
    • Tuberculosis and Respiratory Diseases
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    • v.72 no.3
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    • pp.293-301
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    • 2012
  • Background: Ventilator-associated pneumonia (VAP) requires prompt and appropriate treatment. Since methicillin-resistant Staphylococcus aureus (MRSA) is a frequent pathogen in VAP, rapid identification of it, is pivotal. Our aim was to evaluate the utility of quantitative polymerase chain reaction (qPCR) as a useful method for etiologic diagnoses of MRSA pneumonia. Methods: We performed qPCR for mecA, S. aureus-specific femA-SA, and S. epidermidis-specific femA-SE genes from bronchoalveolar lavage or bronchial washing samples obtained from clinically-suspected VAP. Molecular identification of MRSA was based on the presence of the mecA and femA-SA gene, with the absence of the femA-SE gene. To compensate for the experimental and clinical conditions, we spiked an internal control in the course of DNA extraction. We estimated number of colony-forming units per mL (CFU/mL) of MRSA samples through a standard curve of a serially-diluted reference MRSA strain. We compared the threshold cycle (Ct) value with the microbiologic results of MRSA. Results: We obtained the mecA gene standard curve, which showed the detection limit of the mecA gene to be 100 fg, which corresponds to a copy number of 30. We chose cut-off Ct values of 27.94 (equivalent to $1{\times}10^4$ CFU/mL) and 21.78 (equivalent to $1{\times}10^5$ CFU/mL). The sensitivity and specificity of our assay were 88.9% and 88.9% respectively, when compared with quantitative cultures. Conclusion: Our results were valuable for diagnosing and identifying pathogens involved in VAP. We believe our modified qPCR is an appropriate tool for the rapid diagnosis of clinical pathogens regarding patients in the intensive care unit.

Quantitative Microbial Risk Assessment for Clostridium perfringens in Natural and Processed Cheeses

  • Lee, Heeyoung;Lee, Soomin;Kim, Sejeong;Lee, Jeeyeon;Ha, Jimyeong;Yoon, Yohan
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.8
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    • pp.1188-1196
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    • 2016
  • This study evaluated the risk of Clostridium perfringens (C. perfringens) foodborne illness from natural and processed cheeses. Microbial risk assessment in this study was conducted according to four steps: hazard identification, hazard characterization, exposure assessment, and risk characterization. The hazard identification of C. perfringens on cheese was identified through literature, and dose response models were utilized for hazard characterization of the pathogen. For exposure assessment, the prevalence of C. perfringens, storage temperatures, storage time, and annual amounts of cheese consumption were surveyed. Eventually, a simulation model was developed using the collected data and the simulation result was used to estimate the probability of C. perfringens foodborne illness by cheese consumption with @RISK. C. perfringens was determined to be low risk on cheese based on hazard identification, and the exponential model ($r=1.82{\times}10^{-11}$) was deemed appropriate for hazard characterization. Annual amounts of natural and processed cheese consumption were $12.40{\pm}19.43g$ and $19.46{\pm}14.39g$, respectively. Since the contamination levels of C. perfringens on natural (0.30 Log CFU/g) and processed cheeses (0.45 Log CFU/g) were below the detection limit, the initial contamination levels of natural and processed cheeses were estimated by beta distribution (${\alpha}1=1$, ${\alpha}2=91$; ${\alpha}1=1$, ${\alpha}2=309$)${\times}$uniform distribution (a = 0, b = 2; a = 0, b = 2.8) to be -2.35 and -2.73 Log CFU/g, respectively. Moreover, no growth of C. perfringens was observed for exposure assessment to simulated conditions of distribution and storage. These data were used for risk characterization by a simulation model, and the mean values of the probability of C. perfringens foodborne illness by cheese consumption per person per day for natural and processed cheeses were $9.57{\times}10^{-14}$ and $3.58{\times}10^{-14}$, respectively. These results indicate that probability of C. perfringens foodborne illness by consumption cheese is low, and it can be used to establish microbial criteria for C. perfringens on natural and processed cheeses.

Detection of Viral Hemorrhagic Septicemia Virus (VHSV) from marine fish in the South Western Coastal Area and East China Sea (남.서해안과 동중국해 자연산 어류에서 Viral Hemorrhagic Septicemia Virus(VHSV)검출)

  • Lee, Wol-La;Yun, Hyun-Mi;Kim, Seok-Ryel;Jung, Sung-Ju;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.20 no.3
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    • pp.201-209
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    • 2007
  • Viral hemorrhagic septicemia (VHS) is one of the most serious viral disease of farmed rainbow trout and some marine fishes in Europe and North America. It has been reported in various marine fish species of Asian countries and induced cause mass mortality in Japanese flounder (Paralichthys olivaceus) culturing in Korea. The aims of this study were to monitor VHSV in wild marine fishes and to give critical information for controling the disease through prophylactic methods. Prevalence of the viral disease, geological distribution and reservoir of the virus were investigated using wild marine fishes captured in southern coast and east china sea for two years. (Reverse Transcriptase Polymerase Chain Reaction) RT-PCR results showed that VHSV were detected in 17 (10.6%) out of 160 fish. G gene sequences of viral strains isolated in this study were closely related to that of a reference strain, KVHS01-1, belonging to VHSV genotype Ⅰ. The results suggest that some of wild marine fishes are VHSV carriers and may spread the pathogen directly to fish farmed in coastal area.

Detection of Red Sea Bream Iridovirus (RSIV) from marine fish in the Southern Coastal Area and East China Sea (남.서해안과 동중국해 자연산 어류에서 Red Sea Bream Iridovirus (RSIV)의 검출)

  • Lee, Wol-La;Kim, Seok-Ryel;Yun, Hyun-Mi;Kitamura, Shin Ichi;Jung, Sung-Ju;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.20 no.3
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    • pp.211-220
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    • 2007
  • Red sea bream iridovirus disease (RSIVD) cause massive economic losses in marine aquaculture industry in Korea. The causative agent of this disease (RSIV) infects a wide range of fish species. The aims of this study were to monitor RSIV in wild marine fishes and to give critical information for controling the disease through prophylactic methods. Prevalence of the viral disease, geological distribution and reservoir of the virus were investigated using wild marine fishes captured in southern coast and east china sea for two years. (Polymerase Chain Reaction) PCR results showed that RSIV were detected in 39 (24.3%) out of 160 fish. MCP gene sequences of viral strains isolated in this study were closely related to that of a reference strain, red seabream-K, belonging to Megalocytivirus subgroup Ⅲ. The results suggest that some of wild marine fishes are RSIV carriers and may spread the pathogen directly to fish farmed in coastal area.

Incidence of Citrus vein enation virus in Citrus spp. and Poncirus trifoliata in Korea (국내 감귤류에 발생한Citrus vein enation virus 분포조사)

  • Kim, Bong-Sub;Yang, Hee-Ji;Lee, Su-Hyun;Ko, Seung-Hyun;Park, Kyo Nam;Choi, Eun Jin;Lee, Seong-Jin
    • Research in Plant Disease
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    • v.25 no.4
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    • pp.233-236
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    • 2019
  • Citrus vein enation virus (CVEV), which was regulated as a quarantine virus in Korea, was firstly found on Jeju Island in 2017. In February 2018, a survey was carried out to determine the distribution of CVEV in the main commercial areas growing Citrus spp. and Poncirus trifoliata. The survey was performed at 203 groves in the southern Korean Peninsula and Jeju Island. CVEV infection was determined by reverse transcription polymerase chain reaction detection and sequencing. The coat protein (CP) gene sequences obtained from the CVEV-infected samples showed high similarities (more than 98%) to the previously reported CVEV CP sequences. In summary, CVEV was detected in 136 groves (67%), in which 85.4% of Citrus junos and 77.8% of Citrus unshiu were infected by CVEV. In Jeju Island, the infection rate of CVEV was relatively higher (90.6%). Our result revealed that CVEV has spread widely in Citrus and Poncirus in Korea. Based on the result, the Korean quarantine agency decide to exclude CVEV from quarantine in Korea.

Identification of putative periodontal pathogens in Korean chronic periodontitis patients (한국인 만성 치주염 환자에서 치주질환 원인균의 동정)

  • Yun, Jeong-Ho;Park, Jung-Eun;Kim, Doo-Il;Lee, Syng-Ill;Choi, Seong-Ho;Cho, Kyoo-Sung;Lee, Dae-Sil
    • Journal of Periodontal and Implant Science
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    • v.38 no.2
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    • pp.143-152
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    • 2008
  • Purpose: Specific bacteria are believed to play an important role in chronic periodontitis. Although extensive microbial analyses have been performed from subgingival plaque samples of periodontitis patients, systemic analysis of subingival microbiota has not been carried out in a Korean population so far. The purpose of this study was to investigate the prevalence of 29 putative periodontal pathogens in Korean chronic periodontitis patients and evaluate which pathogens are more associated with Korean chronic periodontitis. Material and Methods: A total of 86 subgingival plaque samples were taken from 15 chronic periodontits(CP) patients and 13 periodontally healthy subjects in Korea. CP samples were obtained from the deepest periodontal pocket (>3 mm probing depth[PD]) and the most shallow periodontal probing site ($\leq$3 mm PD) in anterior tooth and posterior tooth, respectively, of each patient. Samples in healthy subjects were obtained from 1 anterior tooth and 1 posterior tooth. Polymerase chain reaction (PCR) of 16S ribosomal DNA (rDNA) of subgingival plaque bacteria was performed. Detection frequencies(% prevalence) of 29 putative periodontal pathogens were investigated as bacterium-positive sites/total sites. Results: With the exception of Olsenella profuse and Prevotella nigrescens, the sites of diseased patients generally showed higher prevalence than the healthy sites of healthy subjects for all bacteria analyzed. Tanerella forsythensis (B.forsythus), Campylobacter rectus, Filifactor alocis, Fusobacterium nucleatum, Porphyromonas endodontalis and Porphyromonas gingivalis were detected in more than 80% of sites with deep probing depths in CP patients. In comparison between the sites (deep or shallow PD) of CP patients and the healthy sites of healthy subjects, there was statistically significant difference(P<0.05) of prevalence in T.forsythensis (B.forsythus), C.rectus, Dialister invisus, F.alocis, P.gingivalis and Treponema denticola. Conclusion: Our results demonstrate that the four putative periodontal pathogens, T.forsythensis (B.forsythus), C.rectus, P.gingivalis and F.alocis are closely related with CP patients in the Korean population.

Prevalence of major enteric pathogens in different feeding groups of pig in Korean pig farms (국내 양돈장의 사육구간별 주요 소화기질병 원인체 유병율 조사)

  • Jung, Youn-Soo;Park, Yu-Ri;Kang, Dae-Young;Han, Do-Hyun;Yoon, Duhak;Jung, Byeong-Yeal;Park, Choi-Kyu
    • Korean Journal of Veterinary Service
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    • v.39 no.4
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    • pp.211-219
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    • 2016
  • For determining the prevalence of major enteric pathogens, clinical examination and etiological diagnosis were carried out on 75 Korean pig farms. Enteric disease-suspected signs were observed in 90.7% of the farms and the incidence and severity were higher in younger age groups of the pigs. Five of seven pathogens were detected in 375 fecal samples collected from the 75 farms, and the farm-level prevalence of porcine rotavirus group A (PoRVA), pathogenic Escherichia (E.) coli, Lawsonia (L.) intracelluraris, Salmonella spp., and Brachyspira (B.) hyodysenteriae was 54.7%, 54.7%, 16.0%, 10.7% and 2.7%, respectively. PoRVA was extensively infected in suckling and weaning pig groups. The prevalence of pathogenic E. coli was highest in suckling period, and after the period, it exhibited a tendency to decrease. Salmonella spp. and L. intracelluraris were detected in all feeding groups of pigs in a ratio of 1.3~6.7%. B. hyodysenteriae was detected in 1.3~2.7% of growing and fattening pig groups but not detected in suckling and weaning pig groups. At least one or more pathogens were detected in 30.1% of 375 fecal samples. Among these, 25.0% or 5.1% of cases were single or mixed infection. Enteric disease signs of the pigs were significantly co-related with the detection of PoRVA, pathogenic E. coli or Salmonella spp. (P<0.01) but not with L. intracelluraris or B. hyodysenteriae (P>0.05). Conclusively, it will be expected that these data obtained in this study are very useful for subsequent studies and prevention strategies for swine enteric disease in Korean pig farms.

Analysis of Bacterial Wilt Symptoms using Micro Sap Flow Sensor in Tomatoes (식물 생체정보 센서를 활용한 토마토 풋마름병 증상 분석)

  • Ahn, Young Eun;Hong, Kue Hyon;Lee, Kwan Ho;Woo, Young Hoe;Cho, Myeong Cheoul;Lee, Jun Gu;Hwang, Indeok;Ahn, Yul Kyun
    • Journal of Bio-Environment Control
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    • v.28 no.3
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    • pp.212-217
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    • 2019
  • Bacterial wilt caused by Ralstonia solanacearum is a major disease that affects tomato plants widely. R. solanacearum is a soil born pathogen which limits the disease control measures. Therefore, breeding of resistant tomato variety to this disease is important. To identify the susceptible variety, degree of disease resistance has to be determined. In this study, micro sap flow sensor is used for accurate prediction of resistant degree. The sensor is designed to measure sap flow and water use in stems of plants. Using this sensor, the susceptibility to bacterial wilt disease can be identified two to three days prior to the onsite of symptoms after innoculation of R. solanacearum. Thus, this find of diagnosis approach can be utilized for the early detection of bacterial wilt disease.

Safety and Efficacy of Fowl Adenovirus Serotype-4 Inactivated Oil Emulsion Vaccine (닭 유래 아데노 바이러스 혈청형 4형(FAdV-4) 사독 오일 백신의 안전성 및 효능 평가)

  • Kim, Ji-Ye;Kim, Jong-Nyeo;Mo, In-Pil
    • Korean Journal of Poultry Science
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    • v.37 no.3
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    • pp.255-263
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    • 2010
  • Inclusion body hepatitis-hydropericardium syndrome (IBH-HPS) is an acute viral disease usually found in broilers aged from 3 to 5 weeks and causes up to 75% mortality. Among the 12 serotypes of fowl adenovirus group 1, serotype-4 (FAdV-4) was identified as a primary agent of IBH-HPS and was usually isolated in IBH-HPS cases in Korea since 2007. To prevent these IBH-HPS outbreaks in Korea, we developed the FAdV-4 inactivated vaccine using Korean isolate (ADL070244) and evaluated the efficacy of this vaccine. For the efficacy test, 2-week-old specific-pathogen-free (SPF) chickens intramuscularly inoculated with 1 or 2 dose of inactivated vaccine were used and challenged with FAdV-4 through either intramuscular or oral route at 2 weeks after vaccination. The vaccine induced good seroconversion which was confirmed by agar gel precipitation (AGP) test. In addition, the vaccine could decrease the FAdV-4 detection rate and histological lesion severity such as lymphocyte infiltration and necrosis in the liver comparing with those of non-vaccination group. Based on the current results, the developed FAdV-4 inactivated vaccine in this study was effective in the terms of reduction of virus detection rate and histological lesions severity. However, it was difficult to confirm the efficacy of the vaccine clearly because of no mortality and clinical signs in the non-vaccinated group after challenge. Therefore, we need further study to develop a standard challenged model system which could clearly evaluate the efficacy of the vaccines for FAdV-4.