• Pediatric Infection and Vaccine
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• v.30 no.3
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• pp.180-187
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• 2023

In Korea, >90% of children and adolescents aged <19 years have been infected with the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) since 2020. Among confirmed cases of pediatric coronavirus disease 2019 (COVID-19), 40-60% of patients developed neurologic symptoms such as seizures, headache, and encephalitis. Herein, we report the case of a 3-year-old female patient with SARS-CoV-2 infection who presented with seizures and altered consciousness and was diagnosed with COVID-19 encephalitis. The patient recovered after treatment with intravenous immunoglobulin, high-dose steroids, anti-seizure drugs, and an anti-viral agent. She was discharged after regaining the ability to speak words and walk alone on hospital day 39. Complete recovery was observed at the 1-year follow-up. The findings in this case suggest that early detection and active intervention is associated with better outcomes in patients with COVID-19 encephalitis.

• Journal of Microbiology and Biotechnology
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• v.28 no.11
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• pp.1928-1936
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• 2018

Recently, human infections caused by severe fever with thrombocytopenia syndrome virus (SFTSV), which can lead to fatality, have dramatically increased in East Asia. With the unavailability of vaccines or antiviral drugs to prevent and/or treat SFTSV infection, early rapid diagnosis is critical for prevention and control of the disease. Here, we report the development of a simple, rapid and sensitive portable detection method for SFTSV infection applying reverse transcription-loop mediated isothermal amplification (RT-LAMP) combined with one-pot colorimetric visualization and electro-free reaction platform. This method utilizes a pocket warmer to facilitate diagnosis in a resource-limited setting. Specific primers were designed to target the highly-conserved region of L gene of SFTSV. The detection limit of the RT-LAMP assay was approximately $10^0$ viral genome copies from three different SFTSV strains. This assay exhibited comparable sensitivity to qRT-PCR and 10-fold more sensitivity than conventional RT-PCR, with a rapid detection time of 30 to 60 minutes. The RT-LAMP assay using SFTSV clinical specimens has demonstrated a similar detection rate to qRT-PCR and a higher detection rate compared to conventional RT-PCR. Moreover, there was no observed cross-reactive amplification of other human infectious viruses including Japanese Encephalitis Virus (JEV), Dengue, Enterovirus, Zika, Influenza and Middle East Respiratory Syndrome Coronavirus (MERS-CoV). This highly sensitive, electro- and equipment-free rapid colorimetric visualization method is feasible for resource-limited SFTSV field diagnosis.

• Parasites, Hosts and Diseases
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• v.51 no.3
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• pp.269-277
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• 2013

Amoebic keratitis (AK) caused by Acanthamoeba is one of the most serious corneal infections. AK is frequently misdiagnosed initially as viral, bacterial, or fungal keratitis, thus ensuring treatment delays. Accordingly, the early detection of Acanthamoeba would contribute significantly to disease management and selection of an appropriate anti-amoebic therapy. Recently, the loop-mediated isothermal amplification (LAMP) method has been applied to the clinical diagnosis of a range of infectious diseases. Here, we describe a rapid and efficient LAMP-based method targeting Acanthamoeba 18S rDNA gene for the detection of Acanthamoeba using clinical ocular specimens in the diagnosis of AK. Acanthamoeba LAMP assays detected 11 different strains including all AK-associated species. The copy number detection limit for a positive signal was 10 DNA copies of 18S rDNA per reaction. No cross-reactivity with the DNA of fungi or other protozoa was observed. The sensitivity of LAMP assay was higher than those of Nelson primer PCR and JDP primer PCR. In the present study, LAMP assay based on directly heat-treated samples was found to be as efficient at detecting Acanthamoeba as DNA extracted using a commercial kit, whereas PCR was only effective when commercial kit-extracted DNA was used. This study showed that the devised Acanthamoeba LAMP assay could be used to diagnose AK in a simple, sensitive, and specific manner.

• Journal of fish pathology
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• v.23 no.1
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• pp.85-98
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• 2010

Ministry of Agriculture, Fisheries and Food (MAFF) and National Fisheries Research and Development Institute (NFRDI) have inspected the hatchery-reared seeds of 22 marine species and 11 freshwater species for aquatic animal diseases in stock enhancement program in 2009. Results showed that total 12 local selfgovernments have been restocking the sea with cultured juveniles. Gyeongsangnam-do, Jeollanam-do, Jejudo and Chungcheongnam-do have a preference for marine species seeds to freshwater species. On the contrary, freshwater species were released mostly in Gyeonggi-do, Jeollabuk-do and Chungcheongbuk-do. In the marine species group, abalone was the most abundant as 24.5%, and then sea cucumber (15.2%), olive flounder (11.5%), swimming crab (5.6%), black sea bream and rockfish (6.8%), rock bream (5.1%), black rockfish (4.6%) and scorpionfish (4.5%) were followed. Crucian carp was the most abundant as 19.4%, and then eel (17.0%), Korean bullhead (12.3%), melanian snail (12.0%), catfish (8.4%) were followed in the freshwater species group. The total number of inspection cases in this study were 1,080 and disqualification cases were 19 by detection of aquatic animals pathogens such as red sea bream iridovirus (RSIV), koi herpesvirus (KHV) or white spot syndrome virus (WSSV).

• Pediatric Infection and Vaccine
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• v.20 no.2
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• pp.81-88
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• 2013

Purpose : This study was performed to investigate the epidemiology of enterovirus (EV) infections in children at a secondary hospital during recent 5 years. Methods : We collected the cerebrospinal fluid, stool and throat swab samples from the pediatric patients with suspected EV infections in KEPCO Medical Center, Seoul, Korea from July 2006 to September 2010. EV detection and genotype identification were performed by RT-PCR at Korea Centers for Disease Control and Prevention. Results : A total of 386 samples were collected from 277 patients during study period. Ninety-eight patients (35.4%) were diagnosed with EV infections. The RT-PCR positive rate was the highest in throat swab samples (48.3%). The median age of patient was 4.7 years (range, 0.1-12.5 years). Aseptic meningitis (50, 51.0%) was the most common clinical manifestation; herpangina (22, 22.4%) and hand-foot-mouth disease (18, 18.4%). One hundred EVs were isolated from 98 patients and 20 genotypes of EV were identified; Echovirus 30 (28 cases, 28%), Enterovirus 71 (12 cases, 12%), Echovirus 25 (10 cases, 10%), Echovirus 9 (9 cases, 9%) and Coxsackievirus A6 (8 cases, 8%). Aseptic meningitis caused by Echovirus 30 was the most common manifestation in 2008. There was no complicated case caused by Enterovirus 71. Conclusion : This study showed the epidemiology of confirmed EV infection in children from 2006 to 2010. There is a need for continuous surveillance of EV infections and its clinical manifestations.

• Korean Journal of Microbiology
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• v.47 no.2
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• pp.117-123
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• 2011

The purpose of this study was to evaluate and compare different elution and concentration methods for optimization of human rotavirus (HRV) detection method using real-time RT-PCR and cell culture techniques. The leafy vegetable samples (lettuce, Chinese cabbage) were artificially inoculated with HRV. Viruses were extracted from the vegetables by two different elution buffers, buffer A (100 mM Tris-HCl, 50 mM glycine, 3% beef extract, pH 9.5) and buffer B (250 mM Threonine, 300 mM NaCl, pH 9.5), and the extracted viruses were concentrated by filtration and PEG precipitation sequentially. To determine infectivity of the viruses, the viruses recovered from the samples were infected to the MA-104 cells, and integrated cell culture real-time RT-PCR was performed at 1, 48, 72, 96, 120, 144, 168 h post-infection (p.i.). The elution buffer A was more efficient in extracting the virus from the produce samples tested than the buffer B, 29.54% and 18.32% of recoveries, respectively. The sensitivity of real-time RT-PCR method was markedly improved when the virus was concentrated by the filtration method. When the viruses were eluted and concentrated by buffer A and filtration, respectively, the average recovery rate was approximately 51.89%. When the viruses recovered from samples were infected to MA-104 cell, infectious HRV was detected within 48 h p.i. by ICC/real-time RT-PCR, whereas cytopathic effects were not observed until 72 h p.i. The optimized detection method evaluated in this study could be useful for rapid and reliable detection of HRV in fresh produce products and applied for detection of other food-borne viruses.

• 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.

• Korean Journal of Veterinary Research
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• v.38 no.4
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• pp.793-802
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• 1998

Salmonellosis caused by a number of serotypes of Salmonella is an infectious, acute or chronic, zoonotic disease and characterized by enteritis and diarrhea, septicemia in animal. In these studies we investigated the prevalent serotypes of Salmonella causing animal salmonellosis in Korea and the 71 strains of Salmonella spp. were isolated from materials such as mesenteric lymph nodes, fecal samples from slaughtered animal. With the identification test results, the most prevalent serotypes were, in order, S stanley 31 strains (43.7%), S typhimurium 19 strains (26.8%) and S montevideo 11 strains (15.5%), respectively. And we could establish the method for detection of antibodies to broad variety of Salmonella serotypes. Lipopolysaccharide(LPS) antigen extracted from Salmonella was more sensitive and specific than outer membrane protein antigen from that for detection of Salmonella antibody by using an indirect ELISA. The optimal concentration of antigen was 100ng/ml of LPS, the dilutions of conjugate and serum were 1 : 1,000~2,000 and 1 : 200~400, respectively. The mix LPS-ELISA which was used by mixing LPS from S typhimurium (group B), S choleraesuis (group C) and S enteritidis (group D) were more rapid and effective than that used LPS from individual strain for detection of Salmonella serogroup O4, O7 and O9 antibody at the same time. We could obtain the high values of optical density ($0.73{\pm}0.32$) by mix LPS-ELISA on the farm which had occurred salmonellosis, but very low values of $0.17{\pm}0.06$ on the negative farm of salmonellosis. So, the mix LPS-ELISA may be used to monitor the serological surveillance for the presence of infection with a number of serotypes of Salmonella and would be useful for prevention and control of salmonellosis in animal.

• Journal of Korean Society of Environmental Engineers
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• v.29 no.8
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• pp.904-908
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• 2007

The protozoan pathogen Giardia lamblia has been major cause of waterborne enteric disease. In this study, we tried to identify G. lamlbia of human infectious species and to detect viable C. lamblia in river water samples including three sites of Han River mainstream and an its creek using PCR and RT-PCR technique. The PCR/RT-PCR methods were performed by using giardin primer based on the giardin gene targeting ventral disk of Giardia. Sensitivity testing in the DNA/RNA extraction and PCR/RT-PCR amplification steps showed that it was possible to detect a single cyst of G. lamblia and viable G. lamblia. The PCR/RT-PCR methods were compared with immunofluorescence(IF) assay by analyzing 48 samples collected from the mainstream water and the creek water. The mean concentration of the total cysts were 6.3 cysts/10 L(arithmetic mean, n = 48) and the positive detection rate were 62.5%(30/48). And the mean concentration of the cysts excluding empty cysts were 4.5 cysts/10 L and the positive detection rate were 52.1%(25/48). It resulted that 24 of 48 samples included Giardia lamblia by PCR assay and 10 of 48 samples included viable G. lamblia by RT-PCR assay. It resulted that the PCR/RT-PCR technique would be available to river water samples with low concentration of Giardia cysts. And it could support the Korean protozoan standard method, which provides useful information for species and viability.

• Parasites, Hosts and Diseases
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• v.52 no.3
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• pp.305-310
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• 2014

Ascidian soft tunic syndrome (AsSTS) caused by Azumiobodo hoyamushi (A. hoyamushi) is a serious aquaculture problem that results in mass mortality of ascidians. Accordingly, the early and accurate detection of A. hoyamushi would contribute substantially to disease management and prevention of transmission. Recently, the loop-mediated isothermal amplification (LAMP) method was adopted for clinical diagnosis of a range of infectious diseases. Here, the authors describe a rapid and efficient LAMP-based method targeting the 18S rDNA gene for detection of A. hoyamushi using ascidian DNA for the diagnosis of AsSTS. A. hoyamushi LAMP assay amplified the DNA of 0.01 parasites per reaction and detected A. hoyamushi in 10 ng of ascidian DNA. To validate A. hoyamushi 18S rDNA LAMP assays, AsSTS-suspected and non-diseased ascidians were examined by microscopy, PCR, and by using the LAMP assay. When PCR was used as a gold standard, the LAMP assay showed good agreement in terms of sensitivity, positive predictive value (PPV), and negative predictive value (NPV). In the present study, a LAMP assay based on directly heat-treated samples was found to be as efficient as DNA extraction using a commercial kit for detecting A. hoyamushi. Taken together, this study shows the devised A. hoyamushi LAMP assay could be used to diagnose AsSTS in a straightforward, sensitive, and specific manner, that it could be used for forecasting, surveillance, and quarantine of AsSTS.