• Research in Plant Disease
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• v.25 no.1
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• pp.8-15
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• 2019

Plum pox virus (PPV) is a significant viral disease in Prunus spp. worldwide. A nationwide survey was started in Prunus spp. orchards, since PPV was first detected from peach in Korea, 2015. During 2016-2017, samples were collected from 30,333 trees in 1,985 orchards of stone fruits in 8 provinces and 4 cities, Korea and tested by RT-PCR using specific PPV primer set. As a result, 21 trees including peach (9 trees), Japanese apricot (4 trees), plum (1 tree), apricot (7 trees) in 10 orchards were infected and controlled by eradication program. Amplicons of the expected size (547 bp) were obtained from total RNA of seven peach trees in 2016, and directly sequenced. BLAST analysis revealed the highest nucleotide (NT) identity (99%) with a PPV D isolates (LC331298, LT600782) in Genbank. The seven isolates from shared nt sequence identities of 98 to 100% with one another. Phylogenetic analysis showed the isolates in peach clustered closely with the PPV-D isolates from Korea, Japan, USA, and Canada. This is, to our knowledge, the first report of the presence of PPV in Prunus spp. orchards in Korea, 2016-2017, we hope that our results and efforts will contribute to effective measures for eradication of PPV.

• Pediatric Infection and Vaccine
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• v.28 no.2
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• pp.92-100
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• 2021

Purpose: Rapid detection of etiologic organisms is crucial for initiating appropriate therapy in patients with central nervous system (CNS) infection. This study aimed to evaluate the diagnostic value of the BioFire^® Meningitis/Encephalitis (ME) panel in detecting etiologic organisms in cerebrospinal fluid (CSF) samples from febrile infants. Methods: CSF samples from infants aged <90 days who were evaluated for fever were collected between January 2016 and July 2019 at the Seoul National University Children's Hospital. We performed BioFire^® ME panel testing of CSF samples that had been used for CSF analysis and conventional tests (bacterial culture, Xpert^® enterovirus assay, and herpes simplex virus-1 and -2 polymerase chain reaction) and stored at -70℃ until further use. Results: In total, 72 (24 pathogen-identified and 48 pathogen-unidentified) CSF samples were included. Using BioFire^® ME panel testing, 41 (85.4%) of the 48 pathogen-unidentified CSF samples yielded negative results and 22 (91.7%) of the 24 pathogen-identified CSF samples yielded the same results (enterovirus in 19, Streptococcus agalactiae in 2, and Streptococcus pneumoniae in 1) as those obtained using the conventional tests, thereby resulting in an overall agreement of 87.5% (63/72). Six of the 7 pathogen-unidentified samples were positive for human parechovirus (HPeV) via BioFire^® ME panel testing. Conclusions: Compared with the currently available etiologic tests for CNS infection, BioFire^® ME panel testing demonstrated a high agreement score for pathogen-identified samples and enabled HPeV detection in young infants. The clinical utility and cost-effectiveness of BioFire^® ME panel testing in children must be evaluated for its wider application.

• Korean Journal of Microbiology
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• v.44 no.1
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• pp.14-21
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• 2008

Bovine blood, cell, tissue, and organ are used as raw materials for manufacturing biopharmaceuticals, tissue engineered products, and cell therapy. Manufacturing processes for the biologicals using bovine materials have the risk of viral contamination. Therefore viral validation is, essential in ensuring the safety of the products. Bovine herpesvirus type 1 (BHV-1) is the most common bovine pathogen found in bovine blood, cell, tissue, and organ. In order to establish the validation system for the BHV-1 safety of the products, a real-time PCR method was developed for quantitative detection of BHV-1 in raw materials, manufacturing processes, and final products as well as BHV-1 clearance validation. Specific primers for amplification of BHV-1 DNA was selected, and BHV-1 DNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be $2\;TCID_{50}/ml$. The real-time PCR method was validated to be reproducible and very specific to BHV-1. The established real-time PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with BHV-1. BHV-1 DNA could be quantified in CHO cell as well as culture supernatant. Also the real-time PCR assay could detect $10\;TCID_{50}/ml$ of BHV-1 artificially contaminated in bovine collagen. The overall results indicated that this rapid, specific, sensitive, and robust assay can be reliably used for quantitative detection of BHV-1 contamination during the manufacture of biologics.

• Korean Journal of Poultry Science
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• v.18 no.3
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• pp.183-196
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• 1991

In order to establish ELISA method to detect antibody against IBV various factors involved were examined. Antigen was prepared from Massachusetts type IBV which is known to be one of serotypes distributed most widely. The virus was grown in embryonated SPF chicken eggs. Allantoic fluid harvested was processed to ultracentrifugation and sucrose density gradient centrifugation to produce a purified antigen The antisera selected from the field samples based on hemagglutination inhibition test were used as the standard positive and negative sera for this study and the results obtained were summarized as follows. 1 , It was found that ELISA test was satisfactory when the purified antigen was coated on the plate in the amount of about 40ng protein per well. In case of the phospholipase treated hemagglutinating antigen it gave satisfactory results when the each well wns coated with 1.2 to 2.5 hemagglutinating unit which was equivalent to 40 to 90ng of protein. 2. There was no significant difference in the ratio of optical density of positive to that of negative serum whether the coated antigen was held for 1 hour at 37$^{\circ}C$ or it was held overnight at 4$^{\circ}C$. The coated antigen could be kept in dried state without change of antigenecity for at least one month of experimental period at 4$^{\circ}C$. 3. There was a big variation in the optical density and P/N values depending on the maker of the plates and on the plate of the same maker. 4. It was found that background optical density was negligible when serum was diluted more than 1：50 and serum dilution of 1：100 appeared to be appropriate as a routine test dilution to screen the antibody. 5. Optical density was fairly constant 15 minutes afterward from the time substrate was treated and during the 4 hours after stopper was treated. 6. There was a low correlation(r＝0.42) between ELISA and HI test. However, when 74serum samples were tested for the IBV antibody, 98.7% were found to be positive by both tests in which titers of 2$^{6}$ or more by HI test and P/N values of 1.4 or more by ELISA were considered to be positive, 7 Day-old IBV vaccinated chickens shows a similar antibody decay and rising pattern until 8 weeks of age by the two tests, ELISA and HI.

• Clinical and Experimental Pediatrics
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• v.48 no.9
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• pp.976-985
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• 2005

Purpose : Although influenza is one of the most important causes of acute respiratory tract infections in children, effective antiviral therapies are not common and there are only a few clinical studies on treatment of influenza in children. We evaluated the efficacy of oseltamivir in the treatment of naturally aquired influenza in children during the first half of 2004 in Busan. Methods : From January 2004 to June 2004, throat swabs and nasal washes were performed and cultured for the isolation of influenza virus and tested by rapid antigen detection test(QuickVue influenza test) in children with suspected influenza infections. The children who responded positively to the QuickVue influenza test, we divided into two groups : an oseltamivir treatment group and a control group. We compared their clinical symptoms(including fever duration) and diagnosis. The medical records of patients with influenza virus infection were reviewed retrospectively. Results : A total of 621 individuals were suspected of influenza infection. Influenza viruses were isolated in 79(17.2 percent) out of 621 patients examined. QuickVue influenza tests were positive in 181 cases. The treatment group(83 individuals) received oseltamivir twice daily for 5 days, and the control group(99 individuals) were administered only symptom relief medicine. There was no differences between the two groups in clinical diagnosis and symptoms. Oseltamivir treatment reduced the fever duration and other respiratory symptoms. There were no adverse events associated with oseltamivir treatment. Conclusion : Our data suggest that oral oseltamivir treatment reduces the fever duration and other respiratory symptoms of acute influenza without side effects in children.

• Clinical and Experimental Pediatrics
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• v.51 no.8
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• pp.834-841
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• 2008

Purpose : This study was perfomed to analyze in detail the viral etiology of acute lower respiratory tract infections (ALRI) in Cheunan, Korea by multiplex RT-PCR, including human rhinovirus (hRV) and newly identified viruses such as human metapneumovirus (hMPV) and human coronavirus (HCoV-OC43, HCoV-229E/NL63). Method : Nasopharyngeal aspirates (NPA) were collected from 863 hospitalized children with ALRI on the first day of admission at Soonchunhyang University Cheonan Hospital and analyzed by multiplex RT-PCR from December 2005 to November 2006. Results : Viral agents were detected from 474 subjects (54.9%). The identified viral pathogens were hRV 9.2%, hMPV 6.8%, HCoV-229E/NL63 1.4%, and HCoV-OC43 2.1%. Coinfections with ${\geq}2$ viruses were observed in 108 patients (22.8%). The major period of viral ALRI was the first year of life. Clinical diagnoses of viral ALRI were pneumonia (59.5%), bronchiolitis (24.7%), tracheobronchitis (11.4%), and croup (4%). The most common causes of bronchiolitis was respiratory syncytial virus B (RSV B), whereas hMPV, hRV, HCoV-229E/NL63, and HCoV-OC43 were commonly found in patients with pneumonia. The number of hMPV infections peaked between March and May 2006. HCoV-OC43 was prevalent from November to February 2006, whereas HCoV-229E and hRV were detected throughout the year. Conclusion : Although the study was confined to one year, hMPV was not detected during winter and peaked between March and April, which was not consistent with previous studies'. This present study indicates that HCoV is a less common respiratory pathogen in cases of ALRI in Korean children

• Journal of Veterinary Clinics
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• v.29 no.5
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• pp.377-383
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• 2012

Bovine diarrhea is a major economical burden to the bovine industry in Korea. Since multiple infectious agents can be involved in bovine diarrhea, differential diagnosis is essential for effective treatment. Therefore, a panel of two multiplex real-time PCR assays which can simultaneously detect six major bovine enteric pathogens [i.e., bovine viral diarrhea virus (BVDV), bovine coronavirus (BCoV), group A bovine rotavirus (BRV), Salmonella spp., Escherichia coli (E. coli) $K99^+$, and Cryptosporidium parvum] was developed and applied to test 97 fecal samples collected from cattle farms in Korea. In addition, microscopic examination was also preformed on the samples to detect Coccidium oocyst. The estimated sensitivity of the multiplex PCR was 0.1 $TCID_{50}$ for BVDV, BCoV and group A BRV, 5 and 0.5 CFU for E. coli $K99^+$ and Salmonella, respectively, and 50 oocysts for Cryptosporidium. The amplification efficiency of the multiplex PCR ranged between 0.97 and 0.99 for each pathogen. Among 97 samples, 36 samples were positive for at least one of the 6 major pathogens and 6 samples were simultaneously positive for 2 pathogens by the multiplex PCR assay. Coccidium oocysts were also detected in 48 samples, which were all collected from over 1 month old calves. In conclusion, the multiplex real-time PCR panel can be a useful tool for fast and accurate diagnosis of calf diarrhea associated with BVDV, BCoV, group A BRV, E. coli $K99^+$, Salmonella, and/or Cryptosporidium and Coccidium may be an important target which needs to be included in the multiplex PCR panel in the future.

• Korean Journal of Clinical Laboratory Science
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• v.38 no.3
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• pp.196-202
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• 2006

Treatment of hepatitis B virus (HBV) with lamivudine is effective in suppressing virus replication and results in reduced inflammatory activity. However the most troublesome problem of lamivudine treatment is the emergence of lamivudine-resistant strains with amino acid substitution in the YMDD motif of DNA polymerase gene during the treatment. The aim of this study was to determine the mutation of YMDD motif (codon 552) and codon 528 in chronic HBV patients with lamivudine therapy using PCR-direct sequencing and to investigate the relationship between lamivudine mediated HBV mutation and HBeAg. HBV DNA was extracted from serum samples of HBV patients and amplified by nested PCR with two sets of primer pairs selected in HBV DNA polymerase gene. Amplified PCR product was analyzed by 2% agarose gel electrophoresis and direct sequencing. HBV mutation was detected in 124 out of 207 samples (60%). Single mutation was 50.8% for M552I, 43.5% for M552V, 5.7% for M552I/V and the L528M mutation was 67.0%. Double mutation was 43.6% for M552V/L528M, 33.1% for M552I/L528(wild type), 17.7% for M552I/L528M and 5.6% for M552I/V/L528M. Serine mutation at YMDD motif (M552S) was not found and the L528M mutation frequently accompanied M552V type. In this study, the typical difference of frequencies for HBV mutation depending on HBeAg was not found. Moreover, the PCR-direct sequencing method used in this study might be a powerful tool for the mutation study in clinical reference laboratories with high volume.

• Pediatric Infection and Vaccine
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• v.28 no.2
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• pp.101-109
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• 2021

Purpose: Common human coronaviruses (HCoVs) are relatively understudied due to the mild nature of HCoV infection. Given the lack of local epidemiology data on common HCoVs, we aimed to describe clinical and epidemiological characteristics of common HCoVs in children. Methods: Respiratory viral test results from 9,589 respiratory samples from Seoul National University Children's Hospital were analyzed from January 2015 to December 2019. Viral detection was done by the multiplex reverse transcription polymerase chain reaction. Demographics and clinical diagnosis were collected for previously healthy children tested positive for HCoVs. Results: Of the 9,589 samples tested, 1 or more respiratory viruses were detected from 5,017 (52.3%) samples and 463 (4.8%) samples were positive for HCoVs (OC43 2.8%, NL63 1.4%, 229E 0.7%). All 3 types co-circulated during winter months (November to February) with some variation by type. HCoV-OC43 was the most prevalent every winter season. HCoV-NL63 showed alternate peaks in late winter (January to March) and early winter (November to February). HCoV-229E had smaller peaks every other winter. Forty-one percent of HCoV-positive samples were co-detected with additional viruses; human rhinovirus 13.2%, respiratory syncytial virus 13.0%, influenza virus 4.3%. Common clinical diagnosis was upper respiratory tract infection (60.0%) followed by pneumonia (14.8%), croup (8.1%), and bronchiolitis (6.7%). Croup accounted for 17.0% of HCoV-NL63-positive children. Conclusions: This study described clinical and epidemiological characteristics of common HCoVs (OC43, NL63, 229E) in children. Continuing surveillance, perhaps by adding HKU1 in the diagnostic panel can further elucidate the spectrum of common HCoV infections in children.

• Journal of Digital Convergence
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• v.10 no.9
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• pp.1-13
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• 2012

This study aims to analyze actual conditions of the present national defense information network operation, the structure and management of the system, communication lines, security equipments for the lines, the management of network and software, stored data and transferred data and even general vulnerable factors of our army tactical C4I system. Out of them, by carrying out an extensive analysis of the army tactical C4I system, likely to be the core of future information warfare, this study suggested plans adaptive to better information security, based on the vulnerable factors provided. Firstly, by suggesting various information security factor technologies, such as VPN (virtual private network), IPDS (intrusion prevention & detection system) and firewall system against virus and malicious software as well as security operation systems and validation programs, this study provided plans to improve the network, hardware (computer security), communication lines (communication security). Secondly, to prepare against hacking warfare which has been a social issue recently, this study suggested plans to establish countermeasures to increase the efficiency of the army tactical C4I system by investigating possible threats through an analysis of hacking techniques. Thirdly, to establish a more rational and efficient national defense information security system, this study provided a foundation by suggesting several priority factors, such as information security-related institutions and regulations and organization alignment and supplementation. On the basis of the results above, this study came to the following conclusion. To establish a successful information security system, it is essential to compose and operate an efficient 'Integrated Security System' that can detect and promptly cope with intrusion behaviors in real time through various different-type security systems and sustain the component information properly by analyzing intrusion-related information.