• 대한수의학회지
• /
• 제38권2호
• /
• pp.336-344
• /
• 1998

Eight monoclonal antibodies(MAbs) against the transmissible gastroenteritis virus (TGEV) were produced and characterized. Four of the MAbs were produced against a reference TGEV, Purdue strain(P115) and the others were produced against the Korean vaccine virus, Pyungtaek strain. Only one MAb(5C8) produced against P115 had neutralizing activity and was found to be E2 protein-specific. The other seven MAbs(4E2, 4G8, 5H6, 1F8, 2C6, 5H5, and 3A6) had specificity of nucleocapsid protein and no neutralizing activity. All MAbs reacted with different strains of TGEV, but none of the MAbs was reactive with porcine enteropathogenic viruses such as rotavirus, epidemic diarrhea virus and enterovirus by fluorescence antibody(FA) test.

• Clinical and Experimental Pediatrics
• /
• 제55권3호
• /
• pp.77-82
• /
• 2012

Human astrovirus (HAstV) is a major cause of acute diarrhea among children, resulting in outbreaks of diarrhea and occasionally hospitalization. Improved surveillance and application of sensitive molecular diagnostics have further defined the impact of HAstV infections in children. These studies have shown that HAstV infections are clinically milder (diarrhea, vomiting, fever) than infections with other enteric agents. Among the 8 serotypes of HAstV identified, serotype 1 is the predominant strain worldwide. In addition to serotype 1, the detection rate of HAstV types 2 to 8 has increased by using newly developed assays. HAstV is less common compared with other major gastroenteritis viruses, including norovirus and rotavirus; however, it is a potentially important viral etiological agent with a significant role in acute gastroenteritis. A better understanding of the molecular epidemiology and characteristics of HAstV strains may be valuable to develop specific prevention strategies.

• Pediatric Infection and Vaccine
• /
• 제7권1호
• /
• pp.113-119
• /
• 2000

Purpose : Rotavirus is a most common etiologic agent of pediatric gastroenteritis. The standard method to diagnose rotavirus infection was the detection of viral particles in specimens through electron microscopy. But it was complex. Enzyme immunoassay and latex agglutinin are preferred because they are relatively handy, inexpensive and take a short time, in comparison with electron microscopy. However, several reports have shown that the use of ELISA to diagnose rotavirus infection in neonates can result in false positive reactions. The main purpose of this study is to compare ELISA and RT-PCR in the diagnosis of neonatal rotavirus infection. Methods : Data presented in this study were obtained form 123 newborn babies in the nursery of the Fatima Hospital, Masan, Korea, form Jury to December, 1997. We obtained two samples of stool from each of the newborn babies and then performed the Rotazyme test and the RT-PCR. In the Rotazyme test, the results were interpreted according to visual findings. The samples were used for the RT-PCR test after at stock $-30^{\circ}C$ to identify rotavirus group A. The result of the two tests were compared. Results : The informations are divided into 73 males and females. Out of the total informations 15 were transferred from other hospitals. Their average gestational age was $38.5{\pm}1.6$ weeks. The average birth weight was $3134.8{\pm}539gm$. In the Rotazyme test, 75 samples turned out to be positive. Out of them, 55 samples(75.3%) were positive and 18 samples(24.7%) were negative in the RT-PCR. On the other hand, in the Rotazyme test, 50 samples turned out be negative. Out of them, 27 samples(54%) were positive and 23 samples(46%) were negative in the RT-PCR. Conclusion : Rotavirus infection in uncommon in neonates. The diagnosis based on visual findings using Rotazyme test has a disadvantage in the sense that it can result in false positive reactions and false negative reactions in the diagnosis of neonatal rotavirus infection.

• Pediatric Infection and Vaccine
• /
• 제22권2호
• /
• pp.81-90
• /
• 2015

목적: 한국에 백신이 도입된 이후 장염으로 입원한 소아에서 확인된 로타바이러스 유전형의 분포를 알아보고자 하였다. 방법: 2009년 8월부터 2013년 6월 사이에 급성 위장관염으로 입원한 소아에게서 수집된 201개의 로타바이러스 양성 대변 검체를 대상으로 로타바이러스 유전형 분석이 시행되었다. 결과: G 유전형은 G9 (33.3%), G1 (22.4%), G3 (15.9%), G2 (6.0%), G4 (3.0%), G10 (1.5%)이었고 혼합형(15.4%) 및 분류불능(2.5%)이 관찰되었다. P 유전형은 P[4] (45.3%)과 P[8] (43.8%)가 주로 검출되었고 혼합형(10.4%)과 P[2] (0.5%)가 발견되었다. G9P[4] 유전형이 2010/2011년에 자주 검출되었으나 이후에 G1P[8] 유전형이 주로 검출되었다. 결론: 로타바이러스 백신 도입 이후 2009-2013년 동안 G1, G3 및 G9이 번갈아가며 주로 검출되는 것을 확인하였다.

• Pediatric Gastroenterology, Hepatology & Nutrition
• /
• 제9권1호
• /
• pp.1-13
• /
• 2006

목 적: 소아의 급성 위장관염은 매우 흔하고 입원율이 높은 중요한 질환이다. 본 연구에서는 지역 사회에서 소아의 급성 위장관염을 일으키는 대표적인 바이러스 및 세균을 대상으로 1년간 발생 경향에 대한 최근 자료를 제시하고자 하였다. 방 법: 서울대학교 보라매병원 소아과에 3일 이내의 급성 설사를 주소로 내원한 15세 미만의 소아를 대상으로 임상 검사 및 대변 검사를 시행하였다. 입원 소아는 2003년 4월부터 2004년 3월까지 1년간, 외래 소아는 2003년 4월부터 7월까지 단일 외래를 방문한 환아들에 국한하여 연구하였다. 방문 2일 이내에 대변 검체를 얻었으며 바이러스성 원인균 검사로는 로타바이러스, 장 아데노바이러스 항원 검출법을, 세균성 원인균 검사로는 Salmonella, Shigella, Vibrio, Campylobacter 균종 및 Yersinia enterocolitica 대해 배양검사를 시행하였고 병원성 E.coli (ETEC, EHEC, EPEC) 검출을 위해서는 PCR 법을 이용하였다. 입원 환아들은 전해질 검사를 포함한 혈액 검사를 같이 시행받았다. 결 과: 전체 입원 환아는 130명, 외래 환아는 28명이 대상에 포함되었으며, 6세 미만의 환아가 각각 94.6% (123명), 92.8% (26명)로 대부분을 차지하였다. 모두 단일 원인균이 분리되었으며 그 중 로타바이러스가 가장 흔한 원인균으로 입원 환아의 42.3% (55명)에서, 외래 환아의 29.6% (8명)에서 검출되었다. 대변 배양 검사에서는 비장티푸스성 살모넬라균이 입원 환아의 3.9% (4명), 외래 환아의 3.6% (1명)에서 배양되었다. 병원성 대장균은 입원 환아의 2.1% (2명/97명), 외래 환아의 25.0% (3명/12명)에서 분리되었으며 EPEC (4명), ETEC (1명) 순이었고 EHEC는 분리되지 않았다. 이외에 장 아데노바이러스와 Campylobacter, Yersinia, Shigella 균종은 한 예도 없었다. 결 론: 로타바이러스는 국내 지역 사회 병원에서 소아기 위장관염으로 입원하게 되는 가장 흔한 원인균이며 세균성 장염의 원인균으로는 비장티푸스성 살모넬라와 병원성 대장균, 특히 EPEC와 ETEC가 중요하다. Campylobacter 균종은 급성 설사로 입원하게 되는 환아에서는 흔하게 분리되지 않는 것으로 보여지나 외래 환아를 다수 포함한 대단위의 연구가 더 필요하리라 생각된다.

• Journal of Microbiology and Biotechnology
• /
• 제12권3호
• /
• pp.463-469
• /
• 2002

Rotaviruses are the world-wide leading causative agents of severe dehydrating gastroenteritis in young children and animals. The outer capsid glycoprotein VP7 and inner capsid glycoprotein VP6 of rotaviruses are highly antigenic and immunogenic. An SFV-based expression system has recently emerged as a useful tool for heterologous protein production in mammalian cells, exhibiting a much more efficient performance compared to other gene expression systems. Accordingly, the current study adopted an SFV-based expression system to express the VP7 of a group A human rotavirus from a Korean isolate, and the VP6 of a group B bovine rotavirus from a Korean isolate, in mammalian cells. The genes of the VP6 and VP7 were inserted into the SFV expression vector pSFV-1. The RNA was transcribed in vitro from pSFV-VP6 and pSFV-VP7 using SP6 polymerase. Each RNA was then electroporated into BHK-21 cells along with pSFV-helper RNA containing the structural protein gene without the packaging signal. The expression of VP6 and VP7 in the cytoplasm was then detected by immunocytochemistry. The recombinant virus was harvested by ultracentrifugation and examined under electron microscopy. After infecting BHK-21 cells with the defective viruses, the expressed proteins were separated by SDS-PAGE and analyzed by a Western blot. The results indicate that an SFV-based expression system fur the VP6 and VP7 of rotaviruses is an efficient tool for developing a diagnostic kit and/or preventive vaccine.

• Journal of Microbiology and Biotechnology
• /
• 제8권4호
• /
• pp.369-377
• /
• 1998

Rotavirus is a major cause of severe gastroenteritis in young children and animals throughout the world. The VP7 of rotavirus is thought to induce the synthesis of neutralizing antibodies and to be responsible for determining viral serotypes. The cDNA coding for the VP7 capsid protein of human rotavirus, obtained from Korean patients (HRV-Y14), was cloned and its nucleotide sequence was determined. Comparative analysis of the nucleotide sequences between VP7 of Y14 and that of other foreign isolates showed $92.7~95.2\%$ homology to G1 serotypes (RV-4, KU, K8, WA), $74.2\%$ homolgy to G2 serotype HU-5, $76.4\%$ homology to G3 serotype SA-11, and $77.6\%$ homology to G4 serotype A01321. These data suggest that HRV-Y14 can be classified as a G1 serotype. cDNA coding for VP7 of HRV-YI4 was subcloned into the baculovirus vector and the VP7 glycoprotein was expressed in insect cells. The expressed proteins in Sf9 cell extract and tissue culture fluid were separated on SDS-PAGE, and Western blot analysis with monoclonal antibody raised against the synthetic peptide containing 21 amino acids within the VP7 conserved region was performed. The molecular weight of recombinant VP7 was estimated to be 36 kDa which is about the same size as the native VP7. Addition of tunicamycin in the culture media caused a reduction of the molecular weight of the recombinant VP7 indicating that the expressed protein was glycosylated.

• Pediatric Infection and Vaccine
• /
• 제15권2호
• /
• pp.121-128
• /
• 2008

목 적 : 인플루엔자바이러스는 소아에서 급성 호흡기 감염증을 일으키며 주로 겨울철에 유행을 하며, 로타바이러스는 영유아에서 설사를 유발하는 감염성 장염으로 역시 겨울철에 유행하나 2000년대 이후 봄에서 초여름까지 두 바이러스가 분리되어 이에 두 바이러스의 유행 및 임상 양상에 대해 분석하고자 하였다. 방 법 : 2001년 9월부터 2005년 8월까지 한림대학교 의료원에 급성 하기도 감염증을 주소로 입원한 환아를 대상으로 비인두 흡인물을 채취하여 바이러스 배양 및 단일 클론 항체를 이용한 면역 형광 검사법으로 인플루엔자바이러스를 분리한 143례와 같은 기간동안 장염으로 진단 받고 대변 항원 검사에서 로타바이러스 양성인 환아 1,047례의 의무기록을 후향적으로 분석하였다. 결 과 : 호흡기 바이러스 배양 검사를 시행한 3,121명의 비인두 흡인물 중 578명(18.5%)에서 호흡기 바이러스가 배양되었고, 인플루엔자바이러스는 143례로 A형이 96례, B형이 47례였다. 인플루엔자바이러스의 유행 시기는 12월에서 이듬해 6월까지였다. 환아의 남녀 성비는 1.3:1로 남아가 더 많았으며, 중앙연령은 17개월로, 환아의 59%가 24개월 미만에서 발생하였다. 감염시 나타나는 임상증상으로는 기침, 발열, 가래, 콧물 순이었으며 A형과 B형에서 의미있는 차이를 보이지 않았다. 임상진단은 세기관지염 47례(33%), 폐렴 40례(29%), 크룹 25례(18%), 기관기관지염 15례(11%), 합병증을 동반하지 않은 인플루엔자 5례(3%), 중이염 3례(2%), 천식 악화 3례(2%), 부비동염 3례(2%), 열성 경련 1례(1%) 순이었다. 한편 같은 기간동안 설사를 주소로 소아과에 입원한 환아는 3,850명이었으며 이중 1,047명(27%)에서 로타바이러스가 분리되었다. 로타바이러스는 12월에 분리되기 시작하여 3월에 정점을 이룬 뒤 6월까지 분리되었으며 임상증상으로는 설사 912례(87.1%), 구토 768례(73.4%), 발열 496례(47.4%), 두통 52례(5.0%) 순이었다. 결 론 : 인플루엔자바이러스 및 로타바이러스의 유행시기가 한 계절에만 국한되지 않고 연중 분포하는 양상을 보여 이에 대한 감시체계를 좀더 보완하여야 하겠고 예방접종의 시기에 대해서도 고려를 해 보아야 하겠다.

• 한국동물위생학회지
• /
• 제43권4호
• /
• pp.237-244
• /
• 2020

Porcine transmissible gastroenteritis (TGE) has been a significant cause of economic losses in pig farming industry since 1950s. Although transmissible gastroenteritis virus (TGEV) has declined in recent years, it should not be excluded because of its characteristics; the frequency of gene mutation, the mortality in piglets, and the possibility for sudden incidence. Therefore, the herd-level monitoring of the virus is important to prevent further circulation of TGE. The aim of this study is to develop a large-scale sandwich enzyme-linked immunosorbent assay (ELISA) with high specificity to rapidly detect TGEV in feces by using monoclonal antibodies (Mabs). The TGEV specific Mabs were produced in hybridoma cells. Among the Mabs belonged to the IgG class developed by this study, the final selected 8H6, 1B7, 4G3, and 1F8 were identified to have the neutralization ability against TGEV. The sandwich ELISA was established using 8H6 as a reporter antibody and 1B7 and the reported 5C8 as a capture antibody. The developed sandwich ELISA was able to distinguish TGEV from other pathogenic diarrheal agents (porcine rotavirus, porcine reovirus, porcine epidemic diarrhea virus (PEDV), E. coli, and C. perfringens) in tissue culture as well as fecal samples. And the detection rate of TGEV in feces was 80% compared with RT-PCR. The results suggested that the developed sandwich ELISA may be useful in the herd-level monitoring for effective preventive measures due to the early diagnosis of TGEV using a large amount of samples.

• 한국미생물·생명공학회지
• /
• 제47권2호
• /
• pp.317-322
• /
• 2019

Gastroenteritis with diarrhea is one of the most infectious diseases in the world following respiratory infections. Notably, diarrhea-causing viruses (DVs) cause more than 70% of such cases. In this study, 3,065 stool specimens from patients with diarrhea (median age, 1.1 years; range, 0.0-91.1 years), who were admitted to the DanKook University Hospital, were examined using multiplex reverse transcription PCR (mRT-PCR). The target viruses were astrovirus (AstV), enteric adenovirus (EAdV), group A rotavirus (RotV), norovirus GI (NoV-GI), and norovirus GII (NoV-GII). The mRT-PCR results were analyzed based on various factors such as seasonality, age, presence of co-infection, and analyzed trends. The detection rate of the DVs during the study period was found to be 30.8% (n = 943/3,065). When the detection rate was analyzed monthly, the DV detection rate was found to be highest between December to January. Of the detected DVs, NoV-GII was the most common, accounting for 45.5% of the detected viruses (n = 446/980). Notably, 86.5% (n = 848/980) of the pathogens were detected in individuals who were less than 5 years of age. During the study period, NoV-GII and RotV showed alternating trends. In addition, both the number and rate of co-infections increased.