• 제목/요약/키워드: nested-PCR

검색결과 311건 처리시간 0.029초

새우 양식장 주변 생물의 흰점바이러스 보유율 및 두토막눈썹참갯지렁이의 대하 Fenneropenaeus chinensis에 대한 흰점바이러스 전달 효과 (Prevalence of White Spot Syndrome Virus (WSSV) in Marine Organisms around the Shrimp Farm and Polychaete Worm-Mediated Transmission of WSSV to Fenneropenaeus chinensis)

  • 김근식;박상용;이일로;남윤권;방인철
    • 한국어병학회지
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    • 제22권1호
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    • pp.15-21
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    • 2009
  • White spot syndrome virus (WSSV) is a strong causative agent for high mortality in cultured and wild shrimps. From this study, the WSSV prevalence in marine organisms around shrimp farm as well as live feed-mediated transmission of WSSV to farmed shrimps were investigated. Based on nested-PCR method, WSSV was detected in wide array of marine organisms including Perinereis aibuhitensis (81.3% of prevalence rate, 13/16), Enedrias fangi (100%, 16/16), Ruditapes philippinarum (20%, 2/10), crab larvae (100%, 10/10), copepoda (30%, 3/10), Periophthalmus modestus (50%, 5/10), Pachygrapsus crassipes (10%, 1/10), Helice tridens (20%, 2/10) and Neomysis sp. (70%, 7/10). On the other hand, WSSV was not detected in Bullacta exarata, Uca arcuata, and Reishia clavigera. The percent prevalence of WSSV in wild shrimps, Fenneropenaeus chinensis was only 6%, but markedly increased up to 56% after a feeding trial using polychaete worms for one month, indicating that the live feed is one of significant carriers of WSSV to shrimps under practical farming conditions.

Diagnosis and Molecular Analysis on Imported Plasmodium ovale curtisi and P. ovale wallikeri Malaria Cases from West and South Africa during 2013-2016

  • Shin, Hyun-Il;Ku, Bora;Kim, Yu Jung;Kim, Tae Yun;Cho, Shin-Hyeong;Lee, Sang-Eun
    • Parasites, Hosts and Diseases
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    • 제58권1호
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    • pp.61-65
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    • 2020
  • Majority of the imported malaria cases in Korea is attributed to Plasmodium falciparum and P. vivax infections, whereas P. malariae and P. ovale infections are very rare. Falciparum and ovale malaria are mostly imported from Africa, while most of the vivax malaria cases are imported from Southeast Asia. Here, we report 6 Korean imported ovale malaria cases (4 males and 2 females) who had visited in Africa during 2013-2016. These subjects were diagnosed with P. ovale based on microscopic findings, Plasmodium species-specific nested-PCR, and phylogenetic clade using 18S rRNA gene sequences. We identified 2 P. ovale subtypes, 1 P. ovale curtisi (classic type) and 5 P. ovale wallikeri (variant type). All patients were treated with chloroquine and primaquine, and no relapse or recrudescence was reported for 1 year after treatment. With increase of travelers to the countries where existing Plasmodium species, the risk of Plasmodium infection is also increasing. Molecular monitoring for imported malaria parasites should be rigorously and continuously performed to enable diagnosis and certification of Plasmodium spp.

INDUCTION OF MITOCHONDRIAL DNA DELETION BY IONIZING RADIATION IN HUMAN LUNG FIBROBLAST IMR-90 CELLS

  • Eom, Hyeon-Soo;Jung, U-Hee;Park, Hae-Ran;Jo, Sung-Kee
    • Journal of Radiation Protection and Research
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    • 제34권2호
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    • pp.49-54
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    • 2009
  • Mitochondrial DNA (mtDNA) deletion is a well-known marker for oxidative stress and aging and also contributes to their unfavorable effects in cultured cells and animal tissues. This study was conducted to investigate the effect of ionizing radiation (IR) on mtDNA deletion and the involvement of reactive oxygen species (ROS) in this process in human lung fibroblast (IMR-90) cells. Young IMR-90 cells at population doubling (PD) 39 were irradiated with $^{137}Cs$ $\gamma$-rays and the intracellular ROS level was determined by 2',7'-dichlorofluorescein diacetate (DCFH-DA) and mtDNA common deletion (4977bp) was detected by nested PCR. Old cells at PD 55 and $H_2O_2$-treated young cells were compared as the positive control. IR increased the intracellular ROS level and mtDNA 4977 bp deletion in IMR-90 cells dose-dependently. The increases of ROS level and mtDNA deletion were also observed in old cells and $H_2O_2$-treated young cells. To confirm the increased ROS level is essential for mtDNA deletion in irradiated cells, the effects of N-acetylcysteine (NAC) on IRinduced ROS and mtDNA deletion were examined. 5 mM NAC significantly attenuated the IR-induced ROS increase and mtDNA deletion. These results suggest that IR induces the mtDNA deletion and this process is mediated by ROS in IMR-90 cells.

한국 이유자돈에서 검출된 돼지 유행성 설사 바이러스의 유전자 서열 분석 (Genetic sequence analysis of Porcine epidemic diarrhea virus (PEDV) detected from postweaning pigs in Korea)

  • 신현근;김영훈;서태원;한정희
    • 한국동물위생학회지
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    • 제32권1호
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    • pp.11-18
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    • 2009
  • Porcine epidemic diarrhea virus (PEDV), an enveloped single stranded RNA virus in the family Coronaviridae, causes acute viral enteric disease in piglets. Recently outbreaks of porcine epidemic diarrhea (PED) have been rare in Europe but frequent in Asia. In Korea, the increase of PED prevalence is showing specially in postweaning pigs. The purpose of this study was to investigate nucleotide sequence of nucleocapsid protein gene of PEDV field isolates from postweaning pigs in Korea and get more information about the viruses. A total of 15 postweaing pigs clinically suspected of PEDV infection by severe watery diarrhea and dehydration were used in this study. Viral RNA was extracted from small intestines and stools of the pigs. The N gene was amplified by nested RT-PCR, purificated, sequenced, analyzed and then compared with published sequences of other PEDV strains. Three PEDVs were isolated from the suspected postweaning pigs. The N gene of three PEDV field isolates consisted of 483 nucleotides. These PEDV field isolates showed nucleotide sequence homology range from 99.6% to 95% with Chinese strains, from 99.8% to 95.2% with Korean strains, from 97.3% to 95.7% with Japanese strains and from 96.5% to 95.7% with Belgium and British strains. The encoded pritein shared range from 98.8% to 95.6% with Chinese strains, from 99.4% to 95% with Korean strains, from 97.5% to 96.3% with Japanese strains, from 95.6% to 95% with Belgium and British strains. By phylogenetic tree analysis based on nucleotide sequence, three PEDV field isolates were clustered into two groups which were Chinese isolate groups and other Korean isolate groups. These results indicated that some of PEDV field isolates prevailing in Korean postweaning pigs may be associated with those of Chinese strains and other Korean strains.

Molecular Prevalence and Genotypes of Cryptosporidium parvum and Giardia duodenalis in Patients with Acute Diarrhea in Korea, 2013-2016

  • Ma, Da-Won;Lee, Myoung-Ro;Hong, Sung-Hee;Cho, Shin-Hyeong;Lee, Sang-Eun
    • Parasites, Hosts and Diseases
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    • 제57권5호
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    • pp.531-536
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    • 2019
  • Cryptosporidium parvum and Giardia duodenalis are the main diarrhea-causing parasitic pathogens; however, their prevalence in Korea is unknown. Here, we conducted a survey to determine the prevalence and genotype distribution of these 2 pathogens causing acute diarrhea in 8,571 patients hospitalized in 17 Regional Institute of Health Environment sites in Korea, during 2013-2016. C. parvum and G. duodenalis were detected and genotyped by nested PCR, and the isolate were molecularly characterized by sequencing the glycoprotein 60 (Gp60) and ${\beta}-giardin$ genes, respectively. The overall prevalence of C. parvum and G. duodenalis was 0.37% (n=32) and 0.55% (n=47), respectively, and both pathogens were more prevalent in children under 9 years old. Molecular epidemiological analysis showed that the C. parvum isolates belonged to the IIa family and were subtyped as IIaA13G2R1, IIaA14G2R1, IIaA15G2R1, and IIaA18G3R1. Analysis of the ${\beta}-giardin$ gene fragment from G. duodenalis showed that all positive strains belong to assemblage A. This is the first report on the molecular epidemiology and subtyping of C. parvum and G. duodenalis in such a large number of diarrheal patients in Korea. These results highlight the need for continuous monitoring of these zoonotic pathogens and provide a basis for implementing control and prevention strategies. Further, the results might be useful for epidemiological investigation of the source of outbreak.

자연산 대하(Fenneropenaeus chinensis)에서 검출된 노랑머리 바이러스 Genotype 8의 계통분류학적 특성 (Phylogenetic Characteristics of Yellow Head Virus (YHV) Genotype 8 Isolated from Fenneropenaeus chinensis in Korea)

  • 장광일;김보성;오윤경;황지연;권문경;김수미
    • 한국수산과학회지
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    • 제54권5호
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    • pp.698-702
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    • 2021
  • Yellow head virus (YHV) is a rod-shaped positive-sense single-stranded RNA virus, classified into the genus Okavirus, family Roniviridae, and order Nidovirales. In this study, 200 fleshy prawns (Fenneropenaeus chinensis) collected from the vicinity of Narodo in Goheung-gun, Korea, were screened for the presence of yellow head complex viruses and related genotype such as YHV genotype 8. The detection rate of YHV genotype 8 among the 200 fleshy prawns, determined using nested RT-PCR (reverse transcription polymerase chain reation), was 39.0%. Phylogenetic analysis of the ORF1b gene of YHV showed that eight distinct genetic lineages were detected. The four strains of YHV genotype 8 obtained in this study formed a robust clade with the YHV genotype 8 group that was first isolated from fleshy prawns in China suspected to have acute hepatopancreatic necrosis disease (AHPND).

Prevalence and Multilocus Genotyping of Giardia lamblia in Cattle in Jiangxi Province, China: Novel Assemblage E Subtypes Identified

  • Li, Sen;Zou, Yang;Zhang, Xue-Liang;Wang, Ping;Chen, Xiao-Qing;Zhu, Xing-Quan
    • Parasites, Hosts and Diseases
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    • 제58권6호
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    • pp.681-687
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    • 2020
  • Giardia lamblia is a common enteric pathogen associated with diarrheal diseases. There are some reports of G. lamblia infection among different breeds of cattle in recent years worldwide. However, it is yet to know whether cattle in Jiangxi province, southeastern China is infected with G. lamblia. The objectives of the present study were to investigate the prevalence and examine the multilocus genotypes of G. lamblia in cattle in Jiangxi province. A total of 556 fecal samples were collected from 3 cattle breeds (dairy cattle, beef cattle, and buffalo) in Jiangxi province, and the prevalence and genotypes of G. lamblia were determined by the nested PCR amplification of the beta-giardin (bg) gene. A total of 52 samples (9.2%) were positive for G. lamblia. The highest prevalence of G. lamblia was detected in dairy cattle (20.0%), followed by that in beef cattle (6.4%), and meat buffalo (0.9%). Multilocus sequence typing of G. lamblia was performed based on sequences of the bg, triose phosphate isomerase and glutamate dehydrogenase loci, and 22, 42, and 52 samples were amplifiable, respectively, forming 15 MLGs. Moreover, one mixed G. lamblia infection (assemblages A and E) was found in the present study. Altogether, 6 novel assemblage E subtypes (E41*-E46*) were identified for the first time. These results not only provided baseline data for the control of G. lamblia infection in cattle in this southeastern province of China, but also enriched the molecular epidemiological data and genetic diversity of G. lamblia in cattle.

Molecular survey of Toxoplasma gondii B1 gene in pigs from various localities in Korea

  • Dongmi Kwak;Min-Goo Seo
    • Parasites, Hosts and Diseases
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    • 제62권3호
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    • pp.294-301
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    • 2024
  • Toxoplasma gondii, a common protozoan parasite, poses significant public health risks due to its potential to cause toxoplasmosis in humans and can be contracted from pigs, which are considered its critical intermediate host. The aim of this study is to evaluate the prevalence of T. gondii in slaughtered pigs for human consumption, emphasizing the zoonotic implications and the need for improved biosecurity and monitoring practices in pig farming. A total of 1,526 pig samples (1,051 whole blood samples and 384 lung tissue samples from the local slaughterhouse and 91 aborted fetus samples from local farms) were collected throughout the whole country of Korea in 2020. Among them, 6 (0.4%) were found to be infected with T. gondii by nested PCR. When compared by sample type, the prevalence of T. gondii was significantly higher in the aborted fetus samples (2.2%, 2/91) than in the blood (0.3%, 3/1,051) and lung tissue samples (0.3%, 1/384). The B1 gene sequence of T. gondii was similar (97.9-99.8%) to that of the other T. gondii isolates. This study represents the first molecular genotyping survey of T. gondii in the lung tissue of fattening pigs and aborted fetuses in Korea. Our findings indicated the importance of adopting preventive measures including the implementation of rigorous farm hygiene protocols and the promotion of public awareness about the risks of consuming undercooked pork. By addressing the gaps in current control strategies and encouraging the One Health approach, this study contributes to the development of more effective strategies to mitigate the transmission of T. gondii from pigs to humans, ultimately safeguarding public health.

말초 비소세포폐암 환자에서 기관지 세척액 MAGE 유전자 진단의 유용성 (The Utility of MAGE Gene Detection in Bronchial Washing Fluid for Patients with Peripheral NSCLC)

  • 김수현;김호중;권오정;정만표;서지영;고원중;함초롬;남해성;엄상원;권용수;박성훈
    • Tuberculosis and Respiratory Diseases
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    • 제64권1호
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    • pp.15-21
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    • 2008
  • 연구배경: 기관지 내시경으로 조직 검사가 불가능한 고립성 폐결절 환자에서 경피적 폐생검이 중요한 진단 방법으로 사용되고 있으나, 기흉 및 출혈 등의 합병증 및 접근성 등이 문제가 된다. 흑색종항원유전자(Melanoma Antigen Gene, MAGE)는 악성 흑색종에서 처음 발견된 20 종양 항원 및 유전자로서, 비소세포폐암을 비롯한 다양한 암종에서 발현되며 정상 조직에서는 남성의 생식기 세포 및 태반에서만 발현된다. 본 연구는 기관지내시경으로 보이지 않았던 비소세포폐암 환자에서 기관지 세척액을 이용한 MAGE 양성 여부를 검사하여 폐암의 진단에 어느정도 도움이 되는지 알아보고자 하였다. 방법: 2007년 1월부터 2개월간 삼성서울병원 호흡기 내과에서 기관지 내시경을 시행한 환자 중에서 내시경적 조직 검사가 불가능한 고립성 폐결절 환자 37명을 대상으로 하였다. 기관지 세척을 시행하여 MAGE A1-6 RT-nested PCR (iC&G Co, Daegu, Korea)검사와 세포진 검사를 시행하였고, 경피적 폐생검 및 수술을 통한 병리학적인 진단과 비교하였다. 결과: 37명 중에서 비소세포폐암 환자는 21명(56.8%) 이었고(선암종 13/21, 편평상피세포암 8/21), 나머지 16명 (43.2%)은 폐결핵(5), 진균 감염(3), 기질화 폐렴(2) 및 유육종증, 비결핵 항산균 폐질환과 선천성 낭종 등으로 진단되었다. 비소세포폐암 환자에서 세포학적인 검사의 양성률은 9.5% (2/21)에 불과하였으나, MAGE mRNA는 47.6% (10/21)에서 양성을 보여 양성 예측률이 71.4%였다. MAGE 양성률은 편평상피세포암에서 더 높게 나타났다 (선암종 30.8%, 편평상피세포암 75.0%, p>0.05). 양성 병변으로 진단된 16명의 MAGE 발현율은 25.0% (4/16)였다. 결론: 기관지 세척액을 이용한 MAGE 검사는 기관지 내시경을 통한 조직검사가 불가능한 고립성 폐결절에서 기존의 세포진검사에 비해 우수한 양성 예측률을 나타내어 폐암의 진단에 도움이 될 것으로 보인다.

관악산에서 참진드기 조사 및 중증열성혈소판감소증후군 바이러스 검출 (Prevalence of severe fever with thrombocytopenia syndrome virus among ticks surveyed at Mt. Gwanak, Korea)

  • 채정병;김태희;정지호;박윤지;박진호;최경성;유도현;박배근;채준석
    • 대한수의학회지
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    • 제57권3호
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    • pp.169-174
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    • 2017
  • This study was performed to investigate the distribution of ticks and the rate of infection with severe fever with thrombocytopenia syndrome (SFTS) virus in ticks collected at Mt. Gwanak and the Seoul National University campus, Korea. Ticks (n = 273) were collected from May to October and included 76 Haemaphysalis longicornis (4 adult females, 72 nymphs), 49 Haemaphysalis flava (9 adult females, 3 adult males, 37 nymphs), and 148 Haemaphysalis spp. larvae. SFTS virus detection was performed by using one-step RT PCR and nested PCR. The SFTS virus was detected in 7 samples (1 Haemaphysalis longicornis nymph, 3 Haemaphysalis flava nymphs, and 3 Haemaphysalis spp. larva). The overall minimum field infection rate was 2.6%, whereas the minimum field infection rates of adult, nymphal, and larval ticks were 0%, 3.2%, and 2.0%, respectively. For a more accurate indication of the prevalence of SFTS virus in Korea, further in-depth investigations of tick species and SFTS virus occurrence over a larger area and longer period are needed.