• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.16 no.3
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• pp.162-170
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• 2013

Purpose: To detect major acute gastroenteritis virus (rotavirus, norovirus, astrovirus, and enteric adenovirus) and non-enteric type of adenovirus (AdV) in the stools of intussusception patients and to investigate the clinical role of detected viruses. Methods: From March 2012 to February 2013, major acute gastroenteritis virus and non-enteric type of AdV were isolated from stool samples that collected from 44 patients treated for intussusception in Chungnam National University Hospital. Patients were divided according to age and isolated virus. Results: Virus was detected in 28 (63%) stool specimens. The virus detection rate was significantly lower in patients aged under 12 months (p = 0.04). Twenty-two patients (78.6%) had non-enteric adenovirus, 4 (14.3%) had norovirus, 1 (3.6%) had sapovirus, and 1 (3.6%) had astrovirus. AdV subgroup C (AdV 1, 2, 5, and 6) comprised the majority with 20 cases (90.9%). A monthly increment-and-decrement pattern of intussusception was similar to that of viral detection in the stool samples. Enema reductions were successful in 39 patients and surgical manual reductions were performed in 5 patients. Virus was detected in 24 patients (61.5%) of enema reduction group and 4 patients (80.0%) of surgical manual reduction group. All of the detected viruses were non-enteric adenovirus subgroup C (AdV 1, 5, and 6) in surgical reduction patients. Conclusions: The virus detection rate was high in the stools of intussusception patients. The pattern of seasonal intussusception occurrence rate was parallel with seasonal these viral detection rate in the stool samples. These findings suggest that viral infection plays an important role in the development of intussusception and further research is warranted.

• Biomedical Science Letters
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• v.24 no.3
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• pp.230-238
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• 2018

Human enteric Adenovirus-41 (HuEAdV-41) causes gastroenteritis, which detected by the polymerase chain reaction (PCR) base diagnostic system for clinical, food, environmental, fish and shellfish samples. We developed improved PCR and nested PCR primer set which had high specificity, sensitivity and reduced times. In this study, we compared seventeen conditions reported in the previous study that was using the PCR based HuEAdV-41 detection system, and non-enteric Adenovirus were detected in nine conditions. The most sensitive detection condition was up to 25 copies however it took 184 minutes of PCR reaction time. In this study, the PCR primer set developed had same level of sensitivity, it reduced the time of detection for clinical, food and seafood samples to 112 minutes. Developed nested PCR primer set needed 112 minutes but detected up to approximately 1 copy. In addition, developed PCR and nested PCR primer set was validated with twenty samples of underground water at random, of which ten samples showed specific band without non-specific reaction. We expect this study will be used to diagnose HuEAdV-41 from various samples.

• Journal of Life Science
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• v.10 no.2
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• pp.17-20
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• 2000

Adenovirus which is an important infectious viral agent in respiratory and alimentary tract was investigated in Pusan, 1999. Fifteen cases of adenovirus were detected from stools and throat swabs of suspected patients. Two cases of enteric adenovirus were detected from a 5 years old boy and a 6-month-old boy. Thirteen cases of respiratory adenoviruses were detected from children aged under 10 years old and one adult. From respiratory specimens, 1 case of adenovirus type 2, 1 case of type 5, and 11 cases of type 3 were found. Enterotype 41 was detected from fecal preparations. Adenoviruses appeared mostly during winter months, January, February and December. Adenovirus showed a slowly progressive cytopathic effect on HEp-2 cells, Vero cells and BGM cells at 37$^{\circ}C$, in a 5-7% $CO_{2}$ incubation. An electron microscopic observation exhibited non-enveloped icosahedron with a diameter of 70nm. No significant differences on cytopathic effect and morphological features have been found from specimens of either alimentary tract or respiratory secretions.

• Korean Journal of Microbiology
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• v.37 no.4
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• pp.289-293
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• 2001

Incidence of infectious viruses is ensuing throughout the world and threatening the health of children as well as adults. The outbreaks of viral diseases of alimentary tract in Pusan from 1998 to 2000 were detected. Viruses were isolated from stool specimens, cerebrospinal fluid and throat swabs from suspicious patients and confirmed by cell culture, latex agglutination test, indirect immunofluorescent test and electron microscopic observation. The average isolation rate was 12.5% from the suspected specimens. From this work, 2 cases of enteric adenoviruses, 23 cases of echovirus, 31 cases of coxsackivirus 36 cases of rotavirus, 45 cases of SRSV, and 7 cases of poliovirus were detected. The major serotypes of coxsackievirus were B2, B3, B4, B6 and echovirus of serotypes 6, 9, 11, 25, and 30 were examined. Two cases of enteric adenovirus type 41 were also confirmed. The incidence of SRSV was mostly concentrated between December through following March, April through October with echovirus and coxsackievirus, and January through April with rotavirus, respectively. Electron micrograph of negative-stained viruses showed typical appearance with 30-80 nm in diameter.

• Korean Journal of Agricultural Science
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• v.47 no.3
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• pp.673-681
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• 2020

Human enteric Adenovirus 41 (HueAdV-41) is a major waterborne virus that causes human gastroenteritis and is classified as a viral group I double-strand DNA virus, Adenoviridae. HueAdV-41 has been detected with the polymerase chain reaction (PCR) in various samples such as ground water. However, the PCR-based diagnostic method has problems such as reaction time, sensitivity, and specificity. Thus, the loop-mediated isothermal amplification (LAMP) assay has emerged as an excellent method for field applications. In this study, we developed a LAMP system that can rapidly detect HueAdV-41 with high specificity and sensitivity. HueAdV-41 specific LAMP primer sets were tested through a specific, non-specific selection and sensitivity test for three prepared LAMP primer sets, of which only one primer set and optimum reaction temperature were selected. The developed LAMP primer set condition was confirmed as 63℃, and the sensitivity was 1 copy. In addition, to confirm the system, a LAMP positive reaction was developed with the restriction enzyme Taq I (T/GCC). The developed method in this study was more specific, rapid (typically within 2 - 3 hours), and highly sensitive than that of the conventional PCR method. To evaluate and verify the developed LAMP assay, an artificial infection test was done with five cDNAs from groundwater samples, and the results were compared to those of the conventional PCR method. We expect the developed LAMP primer set will be used to diagnose HueAdV-41 from various samples.

• Journal of Microbiology
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• v.44 no.2
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• pp.162-170
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• 2006

Oysters are known to be carriers of food-born diseases, but research on viruses in Korean oysters is scarce despite its importance for public health. We therefore tested oysters cultivated in Goheung, Seosan, Chungmu, and Tongyeong, for viral contamination using cell culture and integrated cell culture PCR (ICC-PCR) with Buffalo green monkey kidney (BGMK) and human lung epithelial (A549) cells. Additional screens via PCR, amplifying viral nucleic acids extracted from oysters supplemented our analysis. Our methods found 23.6 %, 50.9 %, and 89.1 % of all oysters to be positive for adenoviruses when cell culture, ICC-PCR, and direct PCR, respectively, was used to conduct the screen. The same methodology identified enteroviruses in 5.45%, 30.9%, and 10.9% of all cases. Most of the detected enteroviruses (81.3%) were similar to poliovirus type 1; the remainder resembled coxsackievirus type A1. A homology search with the adenoviral sequences revealed similarities to adenovirus subgenera C (type 2, 5, and 6), D (type 44), and F (enteric type 40 and 41). Adenovirus-positive samples were more abundant in A549 cells (47.3%) than in BGMK cells (18.2 %), while the reverse was true for enteroviruses (21.8 % vs. 14.5 %). Our data demonstrate that Korean oysters are heavily contaminated with enteric viruses, which is readily detectable via ICC-PCR using a combination of A549 and BGMK cells.

• Journal of Microbiology and Biotechnology
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• v.18 no.6
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• pp.1156-1163
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• 2008

We performed RT-nested PCR to study the distribution of human enteric viruses in urban rivers in Korea. During 2002-2003, water samples were collected from four rivers in Gyeonggi Province, South Korea. Among 58 samples, 45 (77.6%), 32 (55.2%), 12 (20.7%), 2 (3.4%), 4 (6.9%), and 4 (6.9%) showed positive results with adenoviruses (AdVs), enteroviruses (EVs), reoviruses (ReVs), hepatitis A viruses (HAVs), rotaviruses (RoVs), and sapoviruses (SVs), respectively. According to the binary logistic regression model, the occurrence of each enteric virus, except ReVs and HAVs, was not statistically correlated with the water temperature and levels of fecal coliforms (P<0.05). AdVs were most often detected; only 4 samples (6.9%) were negative for AdVs while positive for other enteric viruses in the studied sites. Our results indicated that monitoring human enteric viruses is necessary to improve microbial quality, and that AdVs detection by PCR can be a useful index for the presence of other enteric viruses in aquatic environments.

• Proceedings of the Microbiological Society of Korea Conference
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• 2008.05a
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• pp.171-171
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• 2008

We monitored the occurrence of human enteric viruses in urban rivers by cell culture-PCR and RT-nested PCR. Water samples were collected monthly or semimonthly between May 2002 and March 2003 in four urban tributaries. Enteric viruses were detected by RT-nested PCR and cell culture-PCR based on a combination of Buffalo Green monkey kidney (BGMK) and A549 cell lines, followed by phylogenetic analysis of amplicons. By RT-nested PCR analysis, 45 (77.6%), 32 (55.2%), 32 (55.2%), 26 (44.8%), 12 (20.7%), 2 (3.4%), 4 (6.9%), and 4 (6.9%) of 58 samples showed positive results with adenoviruses, enteroviruses, noroviruses (NV) genogroup I (GI) and II (GII), reoviruses, hepatitis A viruses, rotaviruses and sapoviruses, respectively. Adenoviruses were most often detected and only eight (13.8%) samples were negative for adenoviruses and positive for other enteric viruses in the studied sites. Thirty-one (77.5%) of the 40 samples were positive for infectious adenoviruses and/or enteroviruses based on cell culture-PCR, and the frequency of positive samples grown on A549 and BGMK (65.0%) was higher than that grown on BGMK alone (47.5%). The occurrence of each enteric virus, except reoviruses and hepatitis A viruses was not statistically correlated with the water temperature and levels of fecal coliforms according to Binary logistic regression model. By sequence analysis, most strains of adenoviruses and enteroviruses detected in this study are similar to the causative agent of viral diseases in Korea and most NV GI- and GII-grouped strains were closely related to the reference strains from China and Japan, and GII/4-related strains had similar sequences to strains recognized as a worldwide epidemic outbreak. Our results suggested that monitoring human enteric viruses is necessary to improve microbial quality and cell culture-PCR using the combination of A549 and BGMK cells and the adenovirus detection by PCR could be useful for monitoring viral contamination in the aquatic environment.

• 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.

• Korean Journal of Microbiology
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• v.38 no.3
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• pp.186-191
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• 2002

The usefulness of oligonucleotide DNA chip was evaluated for detection and primary level identification of major waterborne viruses in environmental samples. The enteric waterborne viruses included enterovirus, adenovirus, and rotavirus. Total intracellular RNA of 10 BGM cell plates showing virus-specific cytopathic effects was extracted at the third day after inoculation. The intracellular RNA was then subjected to either enterovirus-specific RT-PCR followed by sequencing analysis, or the DNA chip. Seven out of 10 positive samples in cell culture were positive but the other three sample were turned out to be negative by both RT-PCR and DNA chip analyses. Nucleotide sequencing results and the DNA chip hybridization results of the RT-PCR product were in complete agreement in the identification of the 7 positive samples as enteroviruses. Using the DNA chip, it took only 3∼4 hr to complete detection and primary level identification of target viruses and additional procedures such as gel electrophoresis or nucleotide sequencing were not necessary. We believe that the DNA chip system can be employed as a highly effective and new detection methodology for environmental viruses.