• 한국식품위생안전성학회지
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• 제30권3호
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• pp.289-294
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• 2015

본 연구에서는 건강기능성 식품원료로서 이용 빈도가 증가하고 있는 하수오, 백수오 및 이엽우피소에 대한 종 특이 프라이머를 개발하였다. 개발된 종 특이 프라이머는 하수오, 백수오 및 이엽우피소에 대해 원물뿐만 아니라 육안 확인이 어려운 가공식품 등을 대상으로 사용원료의 진위여부를 빠르고 정확하게 확인할 수 있었다. 따라서 본 연구에서 개발한 종 특이 PCR방법은 기존의 일반 프라이머를 사용하는 방법이 가지고 있는 식품 적용 한계를 극복할 수 있을 것이라고 판단하였다.

• 한국자원식물학회지
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• 제32권2호
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• pp.201-206
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• 2019

의도적 도입 또는 비의도적 도입에 의해 국내에 확산되고 있는 침입외래식물은 경제적, 환경적인 해를 가할 수 있으며 자생종의 감소나 절멸을 초래하는 등 생물다양성에 부정적인 영향을 미치는 것으로 알려져 있다. 특히 습지나 저수지 및 하안 생육지는 스트레스와 교란에 의해 외래식물 유입과 확산이 용이하므로 국립수목원에서는 이를 관리하기 위한 연구를 지속적으로 수행하고 있다. 본 연구에서는 수원시 하천변에서 발견된 바늘꽃과에 속하는 L. peploides subsp. montevidensis (Spreng.) P.H. Raven가 생태계교란종으로 확산될 우려가 있어 본 종에 대한 기재, 도해, 칼라도판 등을 첨부하여 침입외래식물 관리를 위한 기초자료를 확보하고자 한다.

• Parasites, Hosts and Diseases
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• 제54권3호
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• pp.307-313
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• 2016

Serosurveillance for zoonotic diseases in small mammals and detection of chiggers, the vector of Orientia tsutsugamushi, were conducted from September 2014 to August 2015 in Gwangju Metropolitan Area. Apodemus agrarius was the most commonly collected small mammals (158; 91.8%), followed by Myodes regulus (8; 4.6%), and Crocidura lasiura (6; 3.5%). The highest seroprevalence of small mammals for O. tsutsugamushi (41; 26.3%) was followed by hantaviruses (24; 15.4%), Rickettsia spp. (22; 14.1%), and Leptospira (2; 1.3%). A total of 3,194 chiggers were collected from small mammals, and 1,236 of 3,194 chiggers were identified with 7 species of 3 genera: Leptotrombidium scutellare was the most commonly collected species (585; 47.3%), followed by L. orientale (422; 34.1%), Euchoengastia koreaensis (99; 8.0%), L. palpale (58; 4.7%), L. pallidum (36; 2.9%), Neotrombicula gardellai (28; 2.3%), and L. zetum (8; 0.6%). L. scutellare was the predominant species. Three of 1,236 chigger mites were positive for O. tsutsugamushi by PCR. As a result of phylogenetic analysis, the O. tsutsugamushi strain of chigger mites had sequence homology of 90.1-98.2% with Boryong. This study provides baseline data on the distribution of zoonotic diseases and potential vectors for the development of prevention strategies of vector borne diseases in Gwangju metropolitan area.

• Parasites, Hosts and Diseases
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• 제51권6호
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• pp.689-694
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• 2013

Opisthorchis viverrini and Clonorchis sinensis are parasites known to be carcinogenic and causative agents of cholangiocarcinoma in Asia. The standard method for diagnosis for those parasite infections is stool examination to detect parasite eggs. However, the method has low sensitivity, and eggs of O. viverrini and C. sinensis are difficult to distinguish from each other and from those of some other trematodes. Here, we report a multiplex real-time PCR coupled with high resolution melting (HRM) analysis for the differentiation of O. viverrini and C. sinensis eggs in fecal samples. Using 2 pairs of species-specific primers, DNA sequences from a portion of the mitochondrial NADH dehydrogenase subunit 2 (nad 2) gene, were amplified to generate 209 and 165 bp products for O. viverrini and C. sinensis, respectively. The distinct characteristics of HRM patterns were analyzed, and the melting temperatures peaked at $82.4{\pm}0.09^{\circ}C$ and $85.9{\pm}0.08^{\circ}C$ for O. viverrini and C. sinensis, respectively. This technique was able to detect as few as 1 egg of O. viverrini and 2 eggs of C. sinensis in a 150 mg fecal sample, which is equivalent to 7 and 14 eggs per gram of feces, respectively. The method is species-specific, rapid, simple, and does not require fluorescent probes or post-PCR processing for discrimination of eggs of the 2 species. It offers a new tool for differentiation and detection of Asian liver fluke infections in stool specimens.

• 한국균학회지
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• 제27권2호통권89호
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• pp.132-137
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• 1999

본 연구는 느타리버섯 갈반병 병원균인 pseudomonas tolaasii의 진단을 위한 분자 marker를 개발하기 위해 수행되었다. 세균의 반복염기서열과 펙틴 분해효소 유전자로부터 제작된 여러개의 primer들을 이용해 식용버섯으로부터 분리된 Pseudomonas종들로부터 DNA 다형성을 유도한 바펙틴 분해 효소로부터 제작된 PEU1 primer는 다른 Pseudomonas종들로부터 P. tolaasii를 구분시키는 다형성 밴드를 생성하였다. P. tolaasii 6균주에 공통적으로 나타나는 1.0kb와 0.4kb의 두 가지 밴드를 pGEM-T에 클로닝하고 이들을 각기 pPTOP1과 pPTOP2로 명명하고 probe로 이용하였다. 0.4kb 크기의 pPTOP2의 삽입 DNA는 P. tolaasii 6균주와 hybridization이 이루어진 반면 다른 Pseudomonas종과는 반응하지 않았다. 0.4kb크기의 pPTOP2의 삽입 DNA를 probe로 이용해 dot blot hybridization한 결과 P. tolaasii는 $1.5{\times}10^3\;cfu$까지 검출이 가능함을 확인하였다.

• 한국임상수의학회지
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• 제18권4호
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• pp.329-333
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• 2001

새로 개발되거나 혹은 기존의 어떤 진단검사를 다양한 임상상황에 적용하기 위해서는 먼저 이들 검사법의 진단적 정확도를 추정하는 연구가 반드시 선행되어야 한다. 진단의 정확도에 대한 추정치를 모른다면 검사결과를 해석하는 것이 불가능하기 때문이다. 특히 동일한 개체에서 감염부위별로 3개 이상의 시료를 얻어 진단검사를 적용하는 경우 각 시료의 검사결과는 독립적인 측정시료가 아니라 개체내에서 연관성이 매우 높은 종속적인 시료에 해당 한다. 즉 동일한 개체에서 얻은 시료일수록 검사결과에서 유사한 반응을 보이며 이 경우 분석의 단위는 각각의 개체가 아니라 검사부위가 되는데 이는 의학연구에서 매우 흔하다. 본 연구에서는 Helicobacter 균에 의한 감염을 검출하기 위하여 동일한 개로부터 위의 해부학적 구조상 pyloric antrum, body 및 cardia의 생검시료에 대하여 urease 검사와 PCR 검사를 적용하여 각 검사의 진단적 정확도를 추정하였다. urease 검사의 민감도와 특이도는 0.74 (95% 신뢰구간: 0.64-0.84)와 0.87 (95% 신뢰구간: 0.67-1.00)이었으며 PCR 검사의 민감도와 특이도는 0.95 (95% 신뢰구간: 0.89-1.00)와 0.90 (95% 신뢰구간: 0.70-1.00)로 두 검사의 특이도는 높은 것으로 나타났다. 그러나 PCR 검사에 비하여 urease 검사의 경우 가음성 (false negative)의 가능성이 높기 때문에 진단결과에 대한 신중한 해석이 요구된다.

• 대한본초학회지
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• 제23권1호
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• pp.109-116
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• 2008

Objectives : This study was performed to investigate the discriminative criteria of 6 kinds of Achyranthis Radix by HPLC/DAD. Methods : 20-hydroxyecdysone is isolated by silica gel column chromatography ($CHCl_3$:MeOH, 7:1-1:1 v/v) and identified by nuclear magnetic resonance, A high-performance liquid chromatographic method with diode array detection was used to identify 20-hydroxyecdysone in A. japonica. The analysis was performed using $C_{18}$ column with isocratic elution consisted of 18% acetonitrile and 82% water and the detection was carried out by DAD at 254 nm. 6 kinds of Achyranthis Radix from different locations were extracted in MeOH. Each extracts was analyzed by HPLC in same condition as used in analysis of 20-hydroxyecdysone. The identities of each extracts were determined by comparing the retention time and UV spectrum with that of reference compound. Results : 1. A. japonica and A. bidentata showed the similar patterns of HPLC chromatogram and 20-hydroxycedysone was present in both of them because the peaks having the same retention time and UV spectrum as 20-hydroxyecdysone were shown in the HPLC chromatograms of A. japonica and A. bidentata 2. Cyathula officinalis and C. capitata showed the similar patterns of HPLC chromatogram. The peak having the same retention time and UV spectrum as 20-hydroxyecdysone was shown in the HPLC chromatogram of C. capitata but not shown in the HPLC chromatogram of C. officinalis. 3. Two species of medicinal drugs from Sacheon province showed similar patterns of HPLC chromatogram. Achyranthis Radix from Sacheon(wild) did not have 20-hydroxycedysone but Achyranthis Radix from Sacheon(cultivated) showed the peak having the same retention time as 20-hydroxyecdysone but UV spectrum of the peak was different from that of 20-hydroxyecdysone. Conclusions : These results suggested that 20-hydroxyecdysone could be the discriminative criteria for Achyranthis Radix contain 20-hydroxyecdysone though they belong to different genus and species. And the patterns of HPLC chromatogram also could be the discriminative criteria as the different species of Achyranthis Radix belonging to the same genus showed similar patterns of HPLC chromatogram.

• 한국미생물·생명공학회지
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• 제32권1호
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• pp.91-95
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• 2004

원유로 오염된 토양의 생물학적 복원과정 동안 접종된 Nocardia sp. Hl7-1 균주를 확인하기 위해 165 rDNA sequence에 기초하여 균주에 특이적인 primer를 제작하였다 14균주의 16S rDNA sequence비교를 통해 제작된 4개의 primer set는 Hl7-1 균주를 특이적으로 검출할 수 있었다. 특히 NH169F-NH972R과 NH575F-NH972R의 primer set는 50 fg의 DNA와 $1.2${\times}$10^4$ cfu/g-soil의 균체농도까지 민감하게 검출할 수 있었다. 이 두 primer set는 원유로 오염된 토양의 bioremediation과정 동안 접종된 Hl7-1 균주의 특이적 검출을 가능케 하였으며, 이는 사용된 primer set에 의해 증폭된 PCR산물을 제한효소(EcoRI)로 절단한 결과와 DGGE를 통한 Hl7-1 균주의 확인을 통해 본 연구에서 제작된 primer set의 특이성을 검증하였다.

• Parasites, Hosts and Diseases
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• 제55권5호
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• pp.523-532
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• 2017

A field survey studying intestinal parasites in humans and microbial pathogen contamination at environment was performed in a Laotian rural village to identify potential risks for disease outbreaks. A parasitological investigation was conducted in Ban Lak Sip village, Luang Prabang, Lao PDR involving fecal samples from 305 inhabitants as well as water samples taken from 3 sites of the local stream. Water analysis indicated the presence of several enteric pathogens, i.e., Aeromonas spp., Vibrio spp., E. coli H7, E. coli O157: H7, verocytotoxin-producing E. coli (VTEC), Shigella spp., and enteric adenovirus. The level of microbial pathogens contamination was associated with human activity, with greater levels of contamination found at the downstream site compared to the site at the village and upstream, respectively. Regarding intestinal parasites, the prevalence of helminth and protozoan infections were 68.9% and 27.2%, respectively. Eight helminth taxa were identified in fecal samples, i.e., 2 tapeworm species (Taenia sp. and Hymenolepis diminuta), 1 trematode (Opisthorchis sp.), and 5 nematodes (Ascaris lumbricoides, Trichuris trichiura, Strongyloides stercoralis, trichostrongylids, and hookworms). Six species of intestinal protists were identified, i.e., Blastocystis hominis, Cyclospora spp., Endolimax nana, Entamoeba histolytica/E. dispar, Entamoeba coli, and Giardia lamblia. Questionnaires and interviews were also conducted to determine risk factors of infection. These analyses together with a prevailing infection level suggested that most of villagers were exposed to parasites in a similar degree due to limited socio-economic differences and sharing of similar practices. Limited access to effective public health facilities is also a significant contributing factor.

• 한국어병학회지
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• 제21권3호
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• pp.259-270
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• 2008

Disease surveillance was performed to monitor the prevalence of fish pathogens in wild marine fish caught in coastal offshore water in Korea. A total of 333 of fish samples were collected at set net or fish market at landing port in Pohang (East Sea), Taean (Western Sea), Goseong and Tongyeong (Southern Sea) and 21 species of pathogens causing clinical infections to farmed fish were investigated. The detection rates of fish pathogens from Mugili formes, Tetraodontiformes, Pleuroneciformes, Sorpaeniformes, erciformes and Clupeiformes were 90.9, 61.1, 47.6, 43.6, 37.2 and 11.8%, respectively. Comparing with prevalence of diseases seasonally, both the detection rates of bacteria and parasite were higher than those of virus in April but the detection rates of parasites were distinctively higher than those of bacteria in August with high water temperature. Virus were detected in fish samples caught in the Western and Southern Sea in April. The detected parasites were Trichodina, Ichthyophthirius, Dactylogyrus, Microcotyle, Bivagina, Caligus, Alella and Myxobolus. Among the bacterial pathogens, Vibrio, Streptococcus, Photobacterium, Psuedomonas were predominant. Viral nervous necrosis virus (VNNV) and flounder lymphocystis disease virus (FLDV) were detected from the 6 species of fish virus examined in this study.