• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제16권3호
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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.

• 대한의생명과학회지
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• 제24권3호
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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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• 제10권2호
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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.

• 미생물학회지
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• 제37권4호
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• pp.289-293
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• 2001

감염성 바이러스의 발생은 세계적인 현상으로 어린이는 물론 성인의 건강을 위협하고 있는 설정이다. 1998년-2000년 사이에 부산지역 바이러스성 전염병 유행예측사업의 과정에서 소화기계 바이러스가 탐색되었다. 의심되는 환자의 대변 및 뇌척수액, 인후가검물에서 세포배양, Latex 응집반응, 간접면역형광항체법, 전자현미경 관찰 등을 행하여 바이러스를 확인하였다. 총 검체 중에서 바이러스의 확인 율은 12.5% 이었다. 이 과정을 통하여 3 사례의 장 adenovirus 및 , 23 사례의 echovirus, 31 사례의 coxsackievirus, 36 사례의 rotavirus, 45 사례의 small round structued virus (SRSV), 7 사례의 poliovirus가 확인되었다. 확인된 주요 혈청형으로는 장 adenovirus 41형 및 echovirus 6, 9, 11, 25, 30형, coxsackievirus B2, B3, B4, B6 형 등이 탐색되었다. 각 바이러스의 월별 발생별로는 SRSV는 12월에서는 다음해 4월 사이, echovirus와 coxsackievirus는 4월에서 10월 사이에, rotavirus는 1월에서 4월사이에 각각 분리 율이 높았다. 전자현미경 관찰에서는 30-80 nm의 작은 크기의 바이러스들이 확인되었다.

• 농업과학연구
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• 제47권3호
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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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• 제44권2호
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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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• 제18권6호
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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.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2008년도 International Meeting of the Microbiological Society of Korea
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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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• 제3권1호
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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.

• 미생물학회지
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• 제38권3호
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• pp.186-191
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• 2002

장내바이러스(enterovirus),로타바이러스(rotavirus),그리고 아데노바이러스(adenovirus)등 물을 통해 전파되는 환경바이러스의 신속한 검출 및 1 차적인 분류를 위해 올리고뉴클레오티드 DNA chip을 이용한 분석 시스템의 유용성에 대하여 연구하였다. BGM 세포배양실험에서 바이러스성 세포병변효과(cytopathic effect) 양성으로 판정된 세포단층으로부터 접종배양 3 일 후 세포내 모든 RNA를 분리하여 중합효소연쇄반응과 DNA chip으로 검출여부 및 유전형을 비교 분석한 결과 세포배양에서 양성으로 판정된 10개의 시료 중 3개가 바이러스 음성으로, 7개가 바이러스 양성으로 나타났다. 중합효소연쇄반응과 DNA chip에 의해 양성으로 나타난 7개 시료의 유전형은 두 방법에 의해 모두 장내바이러스로 동정되어 DNA chip에 의한 1차 동정의 유용성도 증명하였다. 세포배양 후 전기영동분석 없이 일차적인 동정과정까지 불과 3∼4 시간 이내에 수행해낼 수 있는 DNA chip분석은 특이성, 신속성, 및 경제성을 고루 갖추어 환경바이러스 검출방법론의 새로운 영역을 구성할 것으로 사료된다.