• 제목/요약/키워드: PCR detection

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Lamivudine 복용 HIV-1 감염자에게서 내성 돌연변이 검색 (Detection of Resistance Mutation to Lamivudine in HIV-1 Infected Patients)

  • 조영걸;성흥섭;이희정;김유겸;지현숙;조군제;강문원
    • 대한미생물학회지
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    • 제35권2호
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    • pp.181-190
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    • 2000
  • To investigate resistance to lamivudine (3TC), we examined the incidence of M184V in 20 HIV-1 patients treated with 3TC for $13.1{\pm}9$ months. Fourteen of 20 patients had been exposed to zidovudine (ZDV) or didanosine (ddI) prior to 3TC therapy. Nested PCR targeting to reverse transcriptase (RT) and direct sequencing were performed for peripheral blood mononuclear cells sampled serially. There were resistance mutations to ZDV in at least 9 patients at baseline, although there was no resistance mutation to 3TC. We could detect M184V in 6 (30%) out of 20 patients. The incidence of M184V increased as the duration of therapy prolongs (13% in samples <12 months; 47% in samples ${\ge}12$ months). The frequency of mutation M184V was higher in patients with previous mutation to ZDV than in patients with wild type. Resistance mutation was not detected in 7 patients. This study shows that resistance to 3TC tends to develop rapidly in patients with baseline mutations or two drugs combination therapy than in those treated simultaneously with triple drugs. This report is the first on resistance to 3TC in Korean AIDS patients.

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Effects of Corydalis Tuber on Synthesis of NO and $PGE_2$ in Murine Macrophage RAW 264.7 Cells Stimulated by LPS

  • Lee, Ki-Young;Park, Se-Keun;Kim, Jeong-Seon;Jang, Mi-Hyeon;Kim, Chang-Ju;Choi, Sun-Mi;Lee, Hye-Jung;Kim, Ee-Hwa
    • 동의생리병리학회지
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    • 제19권3호
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    • pp.785-791
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    • 2005
  • Corydalis Tuber has traditionally been used for the treatment of water retention in the body. Administration of the aqueous extract of Corydalis Tuber has been known to be effective for the control of pain and treatment of arthritis. It was reported that Corydalis Tuber possesses anti-inflammatory activity and modulates the intestinal immune system. The effect of Corydalis Tuber against LPS-stimulated expressions of COX-2, iNOS, and $IL-1{\beta}$ in cells of the murine RAW 264.7 macrophages was investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, reverse transcription-polymerase chain reaction (RT-PCR), $PGE_2$ immunoassay, and NO detection. The aqueous extract of Corydalis Tuber was shown to suppress $PGE_2$ production by inhibition on the LPS-stimulated enhancement of COX-2 enzyme activity, $IL-1{\beta}$, and iNOS expression in the RAW 264.7 macrophages. Present results suggest that Corydalis Tuber exerts anti-inflammatory and analgesic effects probably by suppressing of COX-2, iNOS, and $IL-1{\beta}$ expressions, resulting in inhibition of $PGE_2$ synthesis. Corydalis Tuber has anti-inflammatory and analgesic effects probably by suppressing of COX-2, iNOS, and $IL-1{\beta}$ mRNA expressions, resulting in inhibition of $PGE_2$ and NO synthesis.

Molecular Prevalence of Cryptosporidium spp. among Companion Birds Kept in Pet Shops in Japan

  • Iijima, Yuko;Itoh, Naoyuki;Phrompraphai, Totsapon;Ito, Yoichi;Kimura, Yuya;Kameshima, Satoshi
    • Parasites, Hosts and Diseases
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    • 제56권3호
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    • pp.281-285
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    • 2018
  • Cryptosporidium is the most common protozoan that can infect a wide range of animals, including mammals and birds. Avian Cryptosporidium spp. can cause enteric and respiratory diseases which can be fatal in birds and some species are zoonotic. Companion birds have the potential as reservoir due to their close contact with humans. Pet shops are the major source of companion birds. However, few reports are available regarding Cryptosporidium spp. infection among companion birds kept in pet shops. The present study reports the prevalence and molecular characteristics of Cryptosporidium spp. among companion birds kept in pet shops in Japan. A total of 265 fresh fecal samples were obtained from birds kept in 4 pet shops; these birds belonged to 41 species in 3 bird orders. A nested polymerase chain reaction (PCR) assay targeting the small subunit rRNA gene was employed for the detection of Cryptosporidium spp. A total of 24 samples (9.1%) were positive, and Cryptosporidium spp. were detected from all pet shops. The prevalence of Cryptosporidium spp. in each of the bird orders was 6.5% (10/153) in Psittaciformes, 14.4% (13/90) in Passeriformes, and 4.5% (1/22) in Galliformes. Based on sequence analysis, 13 (54.2%) isolates were classified to C. galli, 8 (33.3%) were avian genotype III, and the remaining 3 (12.5%) were C. baileyi. No infection with zoonotic C. meleagridis and no coinfection with multiple Cryptosporidium spp. and/or genotypes were observed. The zoonotic potential of Cryptosporidium spp. infecting companion birds kept in pet shops in Japan is likely to be low.

국내 가축 유래 대장균에서 CTX-M 및 TEM형 extended-spectrum β-lactamases의 검출 (Detection of CTX-M and TEM type extended-spectrum β-lactamases in Escherichia coli isolated from livestocks in Korea)

  • 조재근;성명숙;김진현;김기석
    • 한국동물위생학회지
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    • 제34권1호
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    • pp.37-43
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    • 2011
  • This study was conducted to investigate the prevalence and genotypes of extended-spectrum ${\beta}$-lactamase (ESBL) in 377 Escherichia coli isolated from healthy and sick animals. Two isolates (0.5%), each of which were isolated from diseased swine and chicken, respectively, were confirmed as ESBL producing isolates by double disk synergy test, and showed a multidrug resistant phenotype. Minimum inhibitory concentration of cefotaxime for the two ESBL producing isolates were 3~4 times higher than those of ceftazidime, respectively. By PCR and sequencing, one isolate from swine have both $bla_{CTX-15}$ and $bla_{TEM-1}$, and one isolate from chicken have $bla_{CTX-15}$ and $bla_{TEM-116}$. Also, these genes were transferred to E. coli J53 by conjugation. These two isolates showed unrelated pulsed-field gel electrophoresis. To our knowledge, this is the first time that $bla_{TEM-116}$ gene was identified in E. coli isolated from animals in Korea. These results suggest more prudent use of third- generation cephalosporins, and surveillance and monitoring for ESBL producing E. coli in both animals and their environments should be necessary.

Reverse Random Amplified Microsatellite Polymorphism Reveals Enhanced Polymorphisms in the 3' End of Simple Sequence Repeats in the Pepper Genome

  • Min, Woong-Ki;Han, Jung-Heon;Kang, Won-Hee;Lee, Heung-Ryul;Kim, Byung-Dong
    • Molecules and Cells
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    • 제26권3호
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    • pp.250-257
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    • 2008
  • Microsatellites or simple sequence repeats (SSR) are widely distributed in eukaryotic genomes and are informative genetic markers. Despite many advantages of SSR markers such as a high degree of allelic polymorphisms, co-dominant inheritance, multi-allelism, and genome-wide coverage in various plant species, they also have shortcomings such as low polymorphic rates between genetically close lines, especially in Capsicum annuum. We developed an alternative technique to SSR by normalizing and alternating anchored primers in random amplified microsatellite polymorphisms (RAMP). This technique, designated reverse random amplified microsatellite polymorphism (rRAMP), allows the detection of nucleotide variation in the 3' region flanking an SSR using normalized anchored and random primer combinations. The reproducibility and frequency of polymorphic loci in rRAMP was vigorously enhanced by translocation of the 5' anchor of repeat sequences to the 3' end position and selective use of moderate arbitrary primers. In our study, the PCR banding pattern of rRAMP was highly dependent on the frequency of repeat motifs and primer combinations with random primers. Linkage analysis showed that rRAMP markers were well scattered on an intra-specific pepper map. Based on these results, we suggest that this technique is useful for studying genetic diversity, molecular fingerprinting, and rapidly constructing molecular maps for diverse plant species.

한국에서 지속감염우의 콧물로부터 소 바이러스성 설사병 바이러스의 계통발생분석 (Phylogenetic Analysis of Bovine Viral Diarrhea Virus from Nasal Swab Sample of Persistently Infected Cattle in Republic of Korea)

  • 송무찬;최경성
    • 한국임상수의학회지
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    • 제26권6호
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    • pp.582-585
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    • 2009
  • 소 바이러스성 설사병 바이러스는 전세계 축산업에서 경제적 파급력이 큰 질병이다. 콧물을 이용한 역전사 중합효소연쇄반응을 이용하여 한우송아지에서 소 바이러스성 설사병 바이러스의 발생이 진단 되었다. 가검물을 채취한 21 마리 소 가운데 12마리가 소 바이러스성 설사병 바이러스에 양성반응을 보여 지속감염우로 분류되었다. 이 동물들은 설사와 폐렴 같은 증상을 나타내었다. 지속감염우는 일발적으로 지속감염 송아지에서 발견된다. 5'-UTR을 이용한 염기서열 및 계통발생 분석에서 본 증례는 BDVD-2a에 속했다. 이들 결과는 콧물을 이용하여 지속감염우를 진단할 수 있다는 것과, 한우 송아지에서 BVDV 근절 대책이 필요하다는 것을 말해 주고 있다.

돼지 group C 로타바이러스 VP6 특이 단클론항체 (Monoclonal antibodies against porcine group C rotavirus VP6)

  • 윤영심;이승철;우상규;조경오;강신영
    • 한국동물위생학회지
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    • 제35권3호
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    • pp.175-182
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    • 2012
  • Rotaviruses have been known to be a major etiological agent of gastroenteritis in both infants and young animals. Subsequently new rotaviruses, which were morphologically indistinguishable but antigenically and electrophoretically distinct with each other, were reported from several animals throughout world including Korea. These new rotaviruses were named as non-group A or group B or group C rotaviruses and so on. It has been very difficult to isolate and grow the non-group A rotaviruses in vitro, and this has greatly limited the characterizations of non-group A rotaviruses and serological studies. In this study, monoclonal antibodies (MAbs) against porcine non-group A rotavirus were produced and characterized. The VP6 gene of porcine group C rotavirus Korean isolate(#06-52-1) was cloned and expressed. For expression of VP6 gene, baculovirus expression system was applied. The VP6 gene and expressed protein in the recombinant virus were confirmed by polymerase chain reaction (PCR), indirect fluorescence antibody (IFA) test and Western blot, respectively. The expressed VP6 was used for MAbs production. The MAbs produced in this study would be promising as diagnostic reagents for detection of group C rotavirus infection.

국내 돼지 유산태아에서 원인 바이러스 검출과 유병률 조사 (Detection and prevalence of viral pathogens from aborted fetuses and stillborn piglets in Korea)

  • 이원광;김성재;김영훈;한정희
    • 한국동물위생학회지
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    • 제34권2호
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    • pp.117-123
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    • 2011
  • This experiment was conducted to investigate the presence of recognized abortifacient viruses from aborted fetuses and stillborn piglets in cases of reproductive failure in sows by PCR. A total of 219 samples of aborted fetuses or stillborn piglets, submitted to the School of Veterinary Medicine of Kangwon National University between 2006 and 2009 May, were collected from 5 provinces in Korea. Abortifacient virus infections were detected in 82 (37.4%) out of 219 aborted fetuses or stillborn piglets as well as on 39 (40.2%) out of 97 pig farms. The major viral infections were porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV) and Aujesky's disease virus (ADV) for which 46 (21%), 19 (8.6%) and 16 (7.3%) were positive, respectively, with 9 fetuses had complicated infection of PCV2+PRRSV or ADV or both. And 8 (3.6%) for SIV, 3 (1.3%) for PPV and 1 (0.4%) for JEV were positive as minor viral infection. The results suggest that PCV2, PRRSV, ADV is apparently the most important viral infectious agents associated with fetal infection leading to abortion or stillbirth in Korea. SIV, PPV and JEV might have a minor impact on reproductive disease.

Improved Resistance to Oxidative Stress by a Loss-of-Function Mutation in the Arabidopsis UGT71C1 Gene

  • Lim, Chae Eun;Choi, Jung Nam;Kim, In A;Lee, Shin Ae;Hwang, Yong-Sic;Lee, Choong Hwan;Lim, Jun
    • Molecules and Cells
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    • 제25권3호
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    • pp.368-375
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    • 2008
  • Approximately 120 UDP-glycosyltransferases (UGTs), which are classified into 14 distinct groups (A to N), have been annotated in the Arabidopsis genome. UGTs catalyze the transfer of sugars to various acceptor molecules including flavonoids. Previously, UGT71C1 was shown to glycosylate the 3-OH of hydroxycinnamates and flavonoids in vitro. Such secondary metabolites are known to play important roles in plant growth and development. To help define the role of UGT71C1 in planta, we investigated its expression patterns, and isolated and characterized a loss-of-function mutation in the UGT71C1 gene (named ugt71c1-1). Our analyses by quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR), microarray data mining, and histochemical detection of GUS activity driven by the UGT71C1 promoter region, revealed the tissue-specific expression patterns of UGT71C1 with highest expression in roots. Interestingly, upon treatment with methyl viologen (MV, paraquat), ugt71c1-1 plants displayed enhanced resistance to oxidative stress, and ROS scavenging activity was higher than normal. Metabolite profiling revealed that the levels of two major glycosides of quercetin and kaempferol were reduced in ugt71c1-1 plants. In addition, when exposed to MV-induced oxidative stress, eight representative ROS response genes were expressed at lower levels in ugt71c1-1 plants, indicating that ugt71c1-1 probably has higher non-enzymatic antioxidant activity. Taken together, our results indicate that ugt71c1-1 has increased resistance to oxidative stress, suggesting that UGT71C1 plays a role in some glycosylation pathways affecting secondary metabolites such as flavonoids in response to oxidative stress.

Identification of Reassortant Pandemic H1N1 Influenza Virus in Korean Pigs

  • Han, Jae-Yeon;Park, Sung-Jun;Kim, Hye-Kwon;Rho, Se-Mi;Nguyen, Giap Van;Song, Dae-Sub;Kang, Bo-Kyu;Moon, Hyung-Jun;Yeom, Min-Joo;Park, Bong-Kyun
    • Journal of Microbiology and Biotechnology
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    • 제22권5호
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    • pp.699-707
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    • 2012
  • Since the 2009 pandemic human H1N1 influenza A virus emerged in April 2009, novel reassortant strains have been identified throughout the world. This paper describes the detection and isolation of reassortant strains associated with human pandemic influenza H1N1 and swine influenza H1N2 (SIV) viruses in swine populations in South Korea. Two influenza H1N2 reassortants were detected, and subtyped by PCR. The strains were isolated using Madin-Darby canine kidney (MDCK) cells, and genetically characterized by phylogenetic analysis for genetic diversity. They consisted of human, avian, and swine virus genes that were originated from the 2009 pandemic H1N1 virus and a neuraminidase (NA) gene from H1N2 SIV previously isolated in North America. This identification of reassortment events in swine farms raises concern that reassortant strains may continuously circulate within swine populations, calling for the further study and surveillance of pandemic H1N1 among swine.