인체 노로바이러스의 한국분리주 Hu/NLV/Gunpo/2006/KO의 분자생물학적 특성

정아용,윤상임,지영미,강윤성,이영민,Jeong, Ah-Yong,Yun, Sang-Im,Jee, Young-Mee,Kang, Yoon-Sung,Lee, Young-Min 한국미생물학회 2009 미생물학회지 Vol.45 No.2

노로바이러스는 급성 위장염을 일으키는 Caliciviridae 과(family)에 속하는 바이러스로 유전자형이 매우 다양하다. 본 연구에서는 노로바이러스 국내분리주의 게놈 RNA로부터 3개의 open reading frame (ORF) 모두의 염기서열을 분석하고, 유전학적 계통분석을 통하여 분자생물학적 특성을 분석하였다. 본 연구에 사용된 노로바이러스(Hu/NLV/Gunpo/2006/KO)는 바이러스성 식중독, 장염 증세를 보이는 2세 여아 가검물로부터 분리되었다. 역전사반응과 PCR 증폭을 통해서 바이러스의 게놈 RNA를 3개의 중첩되는 cDNA 단편으로 합성하였으며, 합성된 cDNA를 염기서열 분석에 직접 사용하였다. 시퀀싱 결과 Hu/NLV/Gunpo/2006/KO는 3개의 ORF (ORF1, 5,100 bp; ORF2, 1,647 bp; ORF3, 765 bp)로 구성되어 있음을 알 수 있었다. 35개의 노로바이러스 국외 분리주와 비교한 결과, ORF1은 ORF2 또는 ORF3에 비해서 상대적으로 염기의 변이율이 낮았으며, 특히 ORF2와 ORF3의 C-말단 부위에서 높은 변이율을 관찰하였다. 유전학적 계통도를 분석한 결과, Hu/NLV/Gunpo/2006/KO는 genogroup II 에 속하며, Saitama U1, Gifu'96, Mc37, Vietnam 026과 같은 클러스터를 형성하는 것을 알 수 있었다. 본 연구를 통하여 노로바이러스 Hu/NLV/Gunpo/2006/KO의 3개의 ORF 염기서열을 모두 밝힘으로써, 앞으로 노로바이러스의 검출법 개발과 유전학적 상관관계뿐 아니라, 유전자의 기능 분석과 관련된 기초연구에 중요한 기초자료를 제공할 수 있을 것으로 기대한다. Norovirus (NV) with a variety of genotypes, a member of the family Caliciviridae, causes acute nonbacterial gastroenteritis in humans. We determined the nucleotide sequence of three open reading frames (ORFs) of a NV Korean strain and characterized the genetic relationship with others. The Korean strain designated Hu/NLV/Gunpo/2006/KO was isolated from the stool specimen of a 2-year-old female suffering from gastroenteritis. By performing reverse transcription and PCR amplification, three overlapping cDNAs were synthesized and used for direct sequencing. We found that like other NVs, this strain contains three ORFs: ORF1, 5,100 bp; ORF2, 1,647 bp; ORF3, 765 bp. Of 35 NVs, ORF1 had a level of genetic diversity lower than ORF2 and ORF3, of which the C-termini of the ORF2 and ORF3 showed a relatively high degree of genetic diversity. Phylogenetic analyses indicated that the Korean strain belonged to genogroup II, with Saitama U1, Gifu'96, Mc37, and Vietnam 026 being formed a single genetic cluster. The nucleotide sequence information of three ORFs of a NV Korean isolate will be useful not only for the development of a diagnostic tool and understanding of genetic relationship, but also provide important basic information for the functional analysis of their gene products.

경기도내 물놀이형 수경시설 중 미생물 분포 조사 연구

정아용(Ah-Yong Jeong),박명기(Myoung-Ki Park),김윤성(Yun-Sung Kim),이창희(Chang-Hee Lee),이정희(Jung-Hee Lee),이혜연(Hye-Yeoun Lee),김영숙(Young-Suk Kim) 한국환경보건학회 2020 한국환경보건학회지 Vol.46 No.6

Objectives: We analyzed water in waterscape facilities to investigate contamination levels of water-borne pathogens and four test items (pH, turbidity, residual chlorine, and Escherichia coli) at facilities including play fountains, splash parks, and artificial streams from June to October in Suwon City and in the whole of Gyeonggido. Methods: A total of 62 waterscape facility samples were collected from 36 sites and tested for pathogenic Escherichia coli and water-borne viruses that cause hand-foot-and-mouth disease, eye disease, and acute enteritis. Results: None of the water-borne pathogens were detected in waterscape facility samples collected from across Gyeonggi-do that were for pre-inspection for facility management. However, the results of samples from Suwon collected in hot weather and during the school vacation period showed five total inconsistencies in turbidity (four cases) and Escherichia coli (one case). Three out of the four inconsistent samples in turbidity were from the same facility which operated a sand filtration system due to its locational factors close to mountains. Conclusion: We suggest that the waterscape facilities in Gyeonggi-do are managed properly in the respect of microbial contamination and water quality.

Real-time PCR을 이용한 환경 중 물 시료의 레지오넬라 분석법 연구

이정희,이정희,박명기,김윤성,윤희정,이창희,정아용,윤미혜 한국환경보건학회 2019 한국환경보건학회지 Vol.45 No.5

Objectives: The standard method for the enumeration of environmental Legionella is culturing, which has several disadvantages, including long incubation and poor sensitivity. The purpose of this study is to demonstrate the usefulness of real-time PCR and to improve the standard method. Methods: In 200 environmental water samples, a real-time PCR and culture were conducted to detect and quantify Legionella. Using with the results of the survey, we compared the real-time PCR with the culture. Results: Each real-time PCR assay had 100% specificity and excellent sensitivity (5 GU/reaction). In the culture, 36 samples were positive and 164 samples were negative. Based on the results of the culture, real-time PCR showed a high negative predictive value of 99%, 35 samples were true positive, 105 samples were true negative, 59 samples were false positive and one sample was a false negative. Quantitative analysis of the two methods indicated a weak linear correlation (r 2 =0.29, r 2 =0.61, respectively). Conclusions: Although it is difficult to directly apply quantitative analysis results of real-time PCR in the enumeration of environmental Legionella, it can be used as a complementary means of culturing to rapidly screen negative samples and to improve the accuracy of diagnosis.

Clinical Characteristics and Etiology of Travelers’ Diarrhea among Korean Travelers Visiting South-East Asia

안지용,정진원,장규진,유명환,채진성,강영아,김성한,정혜숙,천두성,정아용,최은석 대한의학회 2011 Journal of Korean medical science Vol.26 No.2

The morbidity of travelers’ diarrhea (TD) is still high. This study examined the incidence of common pathogens and characteristics of TD among Korean travelers who visited South-East Asian countries. We performed a prospective study involving 479 Korean travelers with diarrheal disease from February 2009 to April 2009 and stool samples were examined and questionnaire surveys were done after arrival. Enterotoxigenic Escherichia coli (ETEC)was found in 36.0% of TD cases, as were the following: Enteroaggregative Escherichia coli (EAEC) in 27.0%, Vibrio parahaemolyticus in 13.1%, and Norovirus in 11.5%. The detected rate of classic TD was higher in men (P = 0.007), in patients who had a shorter duration trip (P = 0.023) and in patients who drank more than 1 liter of water per day (P = 0.037). Positive stool culture rates were higher in men (P = 0.005), in hospitalized patients (P = 0.013). and in those who consumed impure water or raw foods (P = 0.033). A higher severity of disease corresponded to a significantly higher culture positivity rate (P = 0.029). We should consider the possibility of other pathogens in addition to ETEC in patients with TD who visit South-East Asia. Travelers need to educate about risk factors associated with TD.

컬러푸드 중 쿼세틴 및 쿼세틴 배당체 함량조사

박예지(Ye-ji Park),김기철(Ki-Cheol Kim),조상훈(Sang-Hun Cho),문선애(Sun-Ae Moon),김한택(Han-Taek Kim),정아용(Ah-Yong Jeong),김아람(A-Ram Kim),채경석(Kyung-Suk Chae),김지원(Ji-Won Kim),박명기(Myoung-Ki Park) 한국식품영양과학회 2023 한국식품영양과학회지 Vol.52 No.7

본 연구는 국내에서 주로 소비되고 유통되는 컬러푸드의 쿼세틴 및 배당체 함량을 조사하기 위한 것이다. 이를 위해 QuEChERS kit인 ENK1-SC와 WT186008075를 사용하여 추출 및 정제하는 전처리법을 확립하였고, LC-MS/MS를 이용해 21분 동안 negative 모드로 분석하는 기기 분석법을 확립하였다. 서류 1품목, 과일류 4품목, 채소류 8품목(총 100건)의 쿼세틴 및 배당체 함량을 분석한 결과, 브로콜리를 제외한 12품목에 함유되었음을 확인하였다. 총 쿼세틴 함량은 블루베리, 체리, 오디, 토마토, 양파, 포도, 고추, 파프리카, 애호박, 고구마, 콜리플라워, 양배추 순으로 높게 나타났다. Rutin의 함량이 가장 높은 항목은 체리(135.88±7.27μg/g FW)였고, 블루베리에서 hyperin(74.22±14.69 μg/gFW), isoquercitrin(58.48±12.30 μg/g FW), quercitrin(5.47±0.73 μg/g FW)의 함량이 가장 높게 검출되었다. 쿼세틴 함량은 양파(4.96±0.36 μg/g FW)에서 높게 나타났다. 또한, 품목은 같지만 색이 다른 시료의 함량을 비교한 결과, 포도의 isoquercitrin 함량은 청포도보다 약 9배 높게 검출되었고, 청고추의 quercitrin 함량은 홍고추보다 약 8배 높게 나타났다. 쿼세틴 함량이 가장 높은 양파를 부위별로 나누어 함량을 비교한 결과, 과육, 뿌리, 껍질 순으로 isoquercitrin과 쿼세틴의 함량이 증가하였다. 본 연구 결과는 국내 다소비 컬러푸드의 쿼세틴 및 배당체 함량에 대한 영양학적 기초자료로 활용하고자 한다. This study was undertaken to determine the contents of quercetin and quercetin glycosides in colored foods. We developed a sample extraction and separation method using the quick, easy, cheap, effective, rugged, and safe (QuEChERS) kit, and the analytical method was performed by liquid chromatography coupled to mass spectrometry (LC-MS/MS). Totally, 13 colored foods were analyzed; quercetin and quercetin glycosides were obtained in 12 foods, except broccoli. The total quercetin contents of colored foods were high, in the sequential order blueberry, cherry, mulberry, tomato, onion, grape, pepper, sweet pepper, zucchini, sweet potato, cauliflower, and cabbage. Among the colored foods surveyed, the highest content was obtained for rutin, a quercetin glycoside, in cherries (135.88±7.27 μg/g FW). The highest contents obtained in blueberries were hyperin (74.22±14.69 μg/g FW), isoquercitrin (58.48±12.30μg/g FW), and quercitrin (5.47±0.73 μg/g FW). Quercetin content was the highest in onions (4.96±0.36 μg/g FW). Comparing the contents of quercetin and quercetin glycosides according to color, the isoquercitrin content of purple grapes was determined to be about 9 times higher than in green grapes (P<0.05). Furthermore, the contents of rutin, isoquercitrin, and quercitrin were significantly higher in green pepper than in red pepper (P<0.05).