• Pediatric Infection and Vaccine
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• v.3 no.1
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• pp.76-85
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• 1996

Adenoviruses(Ad) are considered to be second only to rotaviruses as the most significant cause of gastroenteritis in young children in Korea and thus it is essential to know the full spectrum of Ad serotypes routinely present in stool specimens from symptomatic patients. Sixty-six Ad isolates and three questionable ones collected over a 2-year peiord were typed by standard microneutralization, restriction endonuclease digestion and PCR of viral DNA to be able to evaluate these assays comprehensively for their ability to identify Ad associated with gastroenteritis. A total of sixty-one isolates(88.4%) were typed: the predominant types were Ad type 41(Ad41)(26.2%), Ad2(19.7%), Ad40(14.8%), Ad5(9.8%), and Ad7(9.8%) which together accounted for almost 80% of the isolates. The remaining virus isolates were typed as Ad1, 31, 34, 3, 25 and a mixture of 40/41. The incidence of Ad31(4.9%) or Ad3(1.6%) was relatively insignificant. DNA restriction analysis(77.5%) proved to be better than serum neutralization but not so when compared to a PCR-based assay for identification of the enteric Ad serotypes(90%) in stool specimens. In this work, the PCR-based assay was evaluated as a tool for the rapid, yet highly sensitive identification of adenoviral DNA sequences in fresh clinical stool specimens.

• The Journal of Korean Society of Virology
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• v.26 no.1
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• pp.23-30
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• 1996

대략 36,000 base pairs (bp)의 두 가닥짜리 DNA를 지놈으로 가진 사람 아데노바이러스 (Ad)는 DNA 상동성(相同性) 및 생물학적/생화학적 성격이 특이한 49개의 혈청형이 알려져 있는데, 이들 대부분의 Ad가 영유아군 및 면역능이 저하된 성인에서 치사적 결과를 초래할 수 있다. Ad의 세포향성(向性)(tropism)은 매우 다양하여 종류에 따라 상기도 감염, 각결막염, 영유아 장염등을 유발하는데 최근 Ad의 다양한 병원성에 대한 원인을 분자생물학적 수준에서 규명하려는 노력의 일환으로 지역에 따라 주되게 출현하는 Ad형 규명이 활발히 이루어지고 있다. Ad 동정/확인은 표면을 이루고 있는 group 공통항원인 hexon 단백질을 탐지하는 효소면역 측정법 (EIA)에 의하며, Ad형별은 Ad fiber의 세포독성 중화시험에의 한다. 그러나, 세포독성 중화시험이 엄청난 노동력 및 시간을 요구하면서도 민감도/특이도가 만족스럽지 못하여 이를 개선하기 위하여 검체 또는 세포배양에서 Ad DNA를 추출하여 제한효소 절단형태를 비교하는 방법이 개발되었는데 이는 세포배양에 잘 자라지 않는 바이러스주의 형별뿐만 아니라 지역 분리주들의 지놈 변형주를 관찰하는 분자생물학적/분자역학적 연구에도 도움이 되고 있다. 국내에는 Ad와 관련된 소아장염의 빈도가 rotavirus에 의한 것 다음으로 빈번한데도 Ad40/41외에 주되게 출현하는 장내 Ad형들이 전혀 규명된 바 없고, 한국형 Ad들의 지놈형태가 전혀 보고된 바가 없다. 또한 세계적으로 Ad형별 조사지역이 늘어감에 따라 유아장염과 연관된 Ad 역시 Ad40, 41이 외의 형들이 Ad40, 41을 능가하는 것으로 보고되고 있는 지역도 있으나 국내에서는 Ad40, 41이외의 형들은 그 역학적 중요도가 전혀 알려져 있지 않다. 이로서 본 연구의 목적은 Ad주들에 특이 중화항체를 이용한 세포독성 중화시험과 Ad DNA 절단법을 적용하여 한국형 장내 Ad주들의 형별을 처음으로 시도함과 동시에 1989-1991사이 출현한 Ad들의 유전적 변형을 관찰하려는 것이었다. 두 방법 모두 사용하였을 때 주되게 출현하는 장내 Ad형들은 Ad4l, Ad2, Ad7, Ad5, 및 Ad40이었다. Ad40/41-양성 검체를 제외한 Ad hexon-EIA양성들의 77.5%를 형별 할 수 있었던 Ad DNA의 제한효소 절단방법은 형들간의 교차중화로 특이성이 낮았던 중화방법 (47.5%)보다 매우 효율적이어서 두 가지 방법을 함께 적응하였을 때는 40주중의 81.5%인 35주를 형별 할 수 있었다. 또한Ad DNA 제한 효소 절단방법은 Ad7 변이주 (Ad7b)도 탐지 할 수 있었다.

• Korean Journal of Microbiology
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• v.37 no.4
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• pp.294-298
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• 2001

Highly effective polymerase chain reaction (PCR) often brings about false positivity caused by contamination of the sample with target nucleic acids. To solve this problem, in situ PCR (ISPCR) has been developed and applied onto various tissue sections and suspension cultures. With combination of PCR and in situ hybridization, this method amplifies the nucleic acid targets in situ and detect the amplified products inside the cells over the background of various cell types. In order to amplify the nucleic acid targets inside the cells, permeabilisation of a sample is required for the entry of amplification reactants into a cell. Treatments of a sample for the purpose allow not only the entry of reactants into the cell but also the exit of amplification products out of the cell. As a means to reduce the leakage of the amplification products, two methods were applied to suspension cultures of HIV-infected Molt/LAV and U 1.1 cells, in which modified, tailed primers produced long linear amplificants whereas biotinylated dUTP instead of dTTP did bulky products.

• Proceedings of the Microbiological Society of Korea Conference
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• 2000.05a
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• pp.94-101
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• 2000

We investigated the viral contamination of water environment including tap water in Korea. River water used for source water was analyzed about monthly between 1997 and 1999 over a period 26 months. A total of 22 tap water samples were collected in 10 sites in 2 urban areas between 1997 and 1998 over a 11 months. All samples were examined for infectious enteroviruses and adenoviruses by a cell culture technique followed by PCR amplification. To identify the recovered viruses from tap water, sequence analysis of PCR products was performed. Infectious viral particles were detected in river water all year round, ranging from 0.93 to 17.3 Most Probable Number of Infectious Unit (MPNIU) /100L. Tap water samples also contained infectious viral particles. The frequency of enteroviruses and adenoviruses in tap water were $50.0\%$ (11/22) and $36.7\%$ (8/22), respectively. Both enteroviruses and adenoviruses were detected in five tap water samples $(22.7\%)$. The level of viral contamination in tap water was quite high, ranging from 0.2 to 2.9 MPNIU/100L, far above the recommended virus level in drinking water set by the U.S. EPA. Poliovirus type 1 derived from vaccine was frequently detected and the remainder comprised coxsackievirus B type or echovirus type 6, which were causative agents of aseptic meningitis in Korea in 1997 and 1998, respectively. Several types of adenovirus were detected in tap water samples and some water samples were found to contain adenoviruses which were closely related to enteric adenovirus type 40 and 41. This stusy shows that surface water and tap water in Korea may be exposed to the risk of viral contamination, especially from recently recognized viruses and this constitutes a potential public health hazard.

• Pediatric Infection and Vaccine
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• v.21 no.3
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• pp.207-213
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• 2014

Purpose: This study aimed to investigate the association between respiratory virus infection and pneumococcal colonization in children. Methods: From May 2009 to June 2010, nasopharyngeal (NP) aspirates were obtained from patients under 18 years old who visited Seoul National University Children's Hospital for respiratory symptoms. NP samples were used to detect respiratory viruses (influenza virus A and B, parainfluenza virus 1, 2 and 3, respiratory syncytial virus A and B, adenovirus, rhinovirus A/B, human metapneumovirus, human coronavirus 229E/NL63 and OC43/HKU1) by RT-PCR and pneumococcus by culture. Results: Median age of the patients was 27 months old. A total of 1,367 NP aspirates were tested for respiratory viruses and pneumococcus. Pneumococcus was isolated from 228 (16.7%) of samples and respiratory viruses were detected from 731 (53.5%). Common viruses were rhinovirus (18.4%), respiratory syncytial virus (RSV) A (10.6%), adenovirus (6.9%), influenza virus A (6.8%). Pneumococcal isolation rate was significantly higher in the cases of positive virus detection than negative detection [21.3% (156/731) vs. 11.3% (72/636), P <0.001]. For individual viruses, pneumococcal isolation rate was positively associated with detection of influenza virus A [24.7% (23/93) vs 16.1% (205/1274), P=0.001], RSV A [28.3% (41/145) vs 15.3% (187/1222), P=0.001], RSV B [31.3% (10/32) vs 16.3% (218/1335), P=0.042], rhinovirus A/B [22.6% (57/252) vs 15.3% (171/1115), P=0.010]. Conclusion: The study revealed that pneumococcal isolation from NP aspirates is related with respiratory virus detection. The result of this study could be used to investigate how respiratory viruses and pneumococcus cause clinical diseases.

• Journal of Dairy Science and Biotechnology
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• v.41 no.3
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• pp.87-102
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• 2023

Globally, acute gastroenteritis is responsible for two million pediatric deaths. In particular, viral gastroenteritis is the most common cause of acute diarrhea, and most children aged <5 years are infected at least once. The common symptoms include profuse watery diarrhea, vomiting, abdominal pain, and fever. Viral gastroenteritis is generally caused by rotavirus, norovirus, astrovirus, and adenovirus. Recently, probiotics use has increased rapidly worldwide due to its inhibitory effect against viral gastroenteritis. In addition, probiotics are known to have anti-inflammatory and anti-allergic effects and enhance immunity without any side effects. Therefore, this review focuses on the anti-viral effects of probiotics on viral gastroenteritis. Furthermore, this review would provide basic data that could be used for developing new products that have improved functionality by addition of probiotics to milk and dairy food.

• Parasites, Hosts and Diseases
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• v.55 no.5
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• pp.513-521
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• 2017

Infectious diarrhea is endemic in most developing countries. We aimed to investigate the protozoan, viral, and bacterial causes of acute diarrhea in Taif, Saudi Arabia. A cross-sectional prospective 1-year study was conducted on 163 diarrheal patients of various ages. Stool samples were collected, 1 per patient, and tested for 3 protozoa, 3 viruses, and 9 bacteria with the Luminex Gastrointestinal Pathogen Panel. Overall, 53.4% (87/163) of samples were positives (20.8% protozoa, 19.6% viruses, 2.8% bacteria, and 9.8% mixed). Rotavirus (19.6%), Giardia duodenalis (16.5%), and Cryptosporidium spp. (8.5%) were the mostly detected pathogens. Adenovirus 40/41 (4.2%), Salmonella (3%), Shiga toxin-producing Escherichia coli (3%), and Entamoeba histolytica (2.4%) were also detected. Norovirus GI/II, Vibrio cholerae, Yersinia enterocolitica, and Clostridium difficile toxin A/B were not detected in any patients. All pathogens were involved in coinfections except E. histolytica. Giardia (5.5%) and rotavirus (3%) were the most commonly detected in co-infections. Enterotoxigenic E. coli (2.4%), Campylobacter spp. (2.4%), E. coli 0157 (1.8%), and Shigella spp. (1.2%) were detected in patients only as co-infections. Infections were more in children 0-4 years, less in adults <40 years, and least >40 years, with statistically significant differences in risk across age groups observed with rotavirus (P<0.001), Giardia (P=0.006), and Cryptosporidium (P=0.036) infections. Lastly, infections were not significantly more in the spring. This report demonstrates the high burden of various enteropathogens in the setting. Further studies are needed to define the impact of these findings on the clinical course of the disease.

• Korean Journal of Microbiology
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• v.39 no.2
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• pp.89-94
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• 2003

Respiratory viruses were isolated from patients with acute respiratory infections in Busan during 2000-2001 and characterized for their antigenic properties. In 2000, 39 out of 43 isolated viruses were identified as influenza viruses and the others were adenoviruses. Among the isolated influenza viruses,23 were type A influenza viruses and 16 were type B influenza viruses. As a result of antigenic characterization, the influenza viruses were determined to A/Sydney/05/97(H3N2)-like, A/Beijing/262/95(H1N1)-like, and B/Harbin07/94-like viruses and serotypes of the isolated adenoviruses were type 1, 2, and 5. In 2001, 56 viruses were isolated and all of the viruses were identified as influenza viruses. They were A/panama/253/99(H3N2)-like and A/Newcaledonia/2007/99(H1Nl)-like viruses when determined by their antigenic properties. The sex distribution of the patients is as follows, 14 males (32.56%),23 females (67.44％) in 2000, and 23 males (41.07%), 33 females (58.93%) in 2001. Occurrence rate was found to be higher in female patients in both years. Age distribution of patients, in 2000, 48.84％ of infection occurred in 0 to 1 year old while in 2002, 33.93% occurred among 11-20 year olds. In 2000, occurrence rate was found to be high in January and again in April and various types of viruses were isolated. These results may be useful for vaccine development and establishment of reliable epidemic data.

• Journal of Periodontal and Implant Science
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• v.41 no.5
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• pp.218-226
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• 2011

Purpose: Micro-computed tomography (micro-CT) has been widely used in the evaluation of regenerated bone tissue but the reliability of micro-CT has not yet been established. This study evaluated the correlation between histomorphometric analysis and micro-CT analysis in performing new bone formation measurement. Methods: Critical-size calvarial defects were created using a 8 mm trephine bur in a total of 24 Sprague-Dawley rats, and collagen gel mixed with autogenous rat bone marrow stromal cells (BMSCs) or autogenous rat BMSCs transduced by adenovirus containing bone morphogenic protein-2 (BMP-2) genes was loaded into the defect site. In the control group, collagen gel alone was loaded into the defect. After 2 and 4 weeks, the animals were euthanized and calvaria containing defects were harvested. Micro-CT analysis and histomorphometric analysis of each sample were accomplished and the statistical evaluation about the correlation between both analyses was performed. Results: New bone formation of the BMP-2 group was greater than that of the other groups at 2 and 4 weeks in both histomorphometric analysis and micro-CT analysis (P=0.026, P=0.034). Histomorphometric analysis of representative sections showed similar results to histomorphometric analysis with a mean value of 3 sections. Measurement of new bone formation was highly correlated between histomorphometric analysis and micro-CT analysis, especially at the low lower threshold level at 2 weeks (adjusted $r^2=0.907$, P<0.001). New bone formation of the BMP-2 group analyzed by micro-CT tended to decline sharply with an increasing lower threshold level, and it was statistically significant (P<0.001). Conclusions: Both histomorphometric analysis and micro-CT analysis were valid methods for measurement of the new bone in rat calvarial defects and the ability to detect the new bone in micro-CT analysis was highly influenced by the threshold level in the BMP-2 group at early stage.

• Nuclear Medicine and Molecular Imaging
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• v.41 no.1
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• pp.54-58
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• 2007

Purpose: Luciferase is one of the most commonly used reporter enzymes in the field of in vivo optical imaging. D-luciferin, the substrate for firefly luciferase has very high cost that allows this kind of experiment limited to small animals such as mice and rats. In this current study, we validated local injection of D-luciferin in the articular capsule for bioluminescence imaging in rabbits. Materials and Methods: Chondrocytes were cultured and infected by replication-defective adenoviral vector encoding firefly luciferase (Fluc). Chondrocytes expressing Fluc were injected or implanted in the left knee joint. The rabbits underwent optical imaging studies after local injection of D-luciferin at 1, 5, 7, 9 days after cellular administration. We sought whether optimal imaging signals was could be by a cooled CCD camera after local injection of D-luciferin. Results: Imaging signal was not observed from the left knee joint after intraperitoneal injection of D-luciferin (15 mg/kg), whereas it was observed after intraarticular injection. Photon intensity from the left knee joint of rabbits was compared between cell injected and implanted groups after intraarticular injection of D-luciferin. During the period of imaging studies, photon intensity of the cell implanted group was 5-10 times higher than that of the cell injected group. Conclusion: We successfully imaged chondrocytes expressing Fluc after intraarticular injection of D-luciferin. This technique may be further applied to develop new drugs for knee joint disease.