• 제목/요약/키워드: infectious cDNA

검색결과 101건 처리시간 0.027초

Monitoring Culicine Mosquitoes (Diptera: Culicidae) as a Vector of Flavivirus in Incheon Metropolitan City and Hwaseong-Si, Gyeonggi-Do, Korea, during 2019

  • Bahk, Young Yil;Park, Seo Hye;Kim-Jeon, Myung-Deok;Oh, Sung-Suck;Jung, Haneul;Jun, Hojong;Kim, Kyung-Ae;Park, Jong Myong;Ahn, Seong Kyu;Lee, Jinyoung;Choi, Eun-Jeong;Moon, Bag-Sou;Gong, Young Woo;Kwon, Mun Ju;Kim, Tong-Soo
    • Parasites, Hosts and Diseases
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    • 제58권5호
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    • pp.551-558
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    • 2020
  • The flaviviruses are small single-stranded RNA viruses that are typically transmitted by mosquitoes or tick vectors and are etiological agents of acute zoonotic infections. The viruses are found around the world and account for significant cases of human diseases. We investigated population of culicine mosquitoes in central region of Korean Peninsula, Incheon Metropolitan City and Hwaseong-si. Aedes vexans nipponii was the most frequently collected mosquitoes (56.5%), followed by Ochlerotatus dorsalis (23.6%), Anopheles spp. (10.9%), and Culex pipiens complex (5.9%). In rural regions of Hwaseong, Aedes vexans nipponii was the highest population (62.9%), followed by Ochlerotatus dorsalis (23.9%) and Anopheles spp. (12.0%). In another rural region of Incheon (habitat of migratory birds), Culex pipiens complex was the highest population (31.4%), followed by Ochlerotatus dorsalis (30.5%), and Aedes vexans vexans (27.5%). Culex pipiens complex was the predominant species in the urban region (84.7%). Culicine mosquitoes were identified at the species level, pooled up to 30 mosquitoes each, and tested for flaviviral RNA using the SYBR Green-based RT-PCR and confirmed by cDNA sequencing. Three of the assayed 2,683 pools (989 pools without Anopheles spp.) were positive for Culex flaviviruses, an insect-specific virus, from Culex pipiens pallens collected at the habitats for migratory birds in Incheon. The maximum likelihood estimation (the estimated number) for Culex pipiens pallens positive for Culex flavivirus was 25. Although viruses responsible for mosquito-borne diseases were not identified, we encourage intensified monitoring and long-term surveillance of both vector and viruses in the interest of global public health.

New Haplotypes of the ATP Synthase Subunit 6 Gene of Mitochondrial DNA are Associated with Acute Lymphoblastic Leukemia in Saudi Arabia

  • Yacoub, Haitham Ahmed;Mahmoud, Wael Mahmoud;El-Baz, Hatim Alaa-Eldeen El-Din;Eid, Ola Mohamed;El-Fayoumi, Refaat Ibrahim;Mahmoud, Maged Mostafa;Harakeh, Steve;Abuzinadah, Osama H.A.
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권23호
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    • pp.10433-10438
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    • 2015
  • Background: Acute lymphoblastic leukemia (ALL) is the most common cancer diagnosed in children and represents approximately 25% of cancer diagnoses among those younger than 15 years of age. Aim and Objectives: This study investigated substitutions in the ATP synthase subunit 6 gene of mitochondrial DNA (mtDNA) as a potential diagnostic biomarker for early detection and diagnosis of acute lymphoblastic leukemia. Based on mtDNA from 23 subjects diagnosed with acute lymphoblastic leukemia, approximately 465 bp of the ATP synthase subunit 6 gene were amplified and sequenced. Results: The sequencing revealed thirty-one mutations at 14 locations in ATP synthase subunit 6 of mtDNA in the ALL subjects. All were identified as single nucleotide polymorphisms (SNPs) with a homoplasmic pattern. The mutations were distributed between males and females. Novel haplotypes were identified in this investigation: haplotype (G) was recorded in 34% in diagnosed subjects; the second haplotype was (C) with frequency of 13% in ALL subjects. Neither of these were observed in control samples. Conclusions: These haplotypes were identified for the first time in acute lymphoblastic leukemia patients. Five mutations able to change amino acid synthesis for the ATP synthase subunit 6 were associated with acute lymphoblastic leukemia. This investigation could be used to provide an overview of incidence frequency of acute lyphoblastic leukemia (ALL) in Saudi patients based on molecular events.

한국에서 분리된 IHNV-PRT의 G protein의 유전자 클로닝과 염기서열 분석 (Molecular Cloning and Nucleotide Sequence of the G protein of a Korean Isolate of Infectious Hematopoietic Necrosis Virus)

  • 김영조;허강준;박정우;박정문
    • 미생물학회지
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    • 제35권3호
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    • pp.226-230
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    • 1999
  • 무지개송어 등의 연어과 어류에서 발생하는 전염성 조혈기괴사증 바이러스의 국내분리주인 IHNV-PRT 의 특성을 규명하기 위하여 IHNV-PRT 의 당단백질인 G를 암호화하고 있는 유전자의 일부를 PCR로 증폭한 후 cDNA를 클로닝하여 이들의 염기서열을 분석하였다. 이 PCR product 는 442 bp의 크기이었다. IHNV-PRT 의 G의 염기서열을 IHNV 의 다른 strain 과 비교 분석한 결과 IHNV-RB-76, IHNV-RB, IHNV-LR-73, IHNV-K, IHNV-WRAC, IHNV-SRCV, IHNV-CoI-85들의 G와 각각 95,94,94,94,93,93%의 상동성을 보였다. 그러나 넙치로부터 분리된 fish rhabdovirus 인 hirame rhabdovirus 의 G와는 81%의 사동성을 보였다. 이 결과로부터 IHNV 의 G 유전자는 비록 HRV 의 G 유전자와 유사성은 높지 않지만, IHNV 의 strain 에 관계없이 유사하다는 것을 알 수 있다.

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Identification and Antibiotic Susceptibility of the Bacteria from Non-odontogenic Infectious Lesions

  • Kim, Yong Min;Kim, Jae-Jin;Kim, Mija;Park, Soon-Nang;Kim, Hwa-Sook;Kook, Joong-Ki;Kim, Hak Kyun
    • International Journal of Oral Biology
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    • 제39권2호
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    • pp.87-95
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    • 2014
  • The purpose of this study was to isolate and identify bacteria from the 4 patients with non-odontogenic infectious lesions (mucormycosis, chronic inflammation from wound infection, and two actinomycosis) and determine their antimicrobial susceptibility against eight antibiotics. Bacterial culture was performed under three culture conditions (anaerobic, $CO_2$, and aerobic incubator). The bacterial strains were identified by 16S rRNA gene (16S rDNA) sequence comparison analysis method. For investigating the antimicrobial susceptibility of the bacteria against eight antibiotics, penicillin G, amoxicillin, tetracycline, cefuroxime, erythromycin, clindamycin, vancomycin, and Augmentin$^{(R)}$ (amoxicillin + clavulanic acid), minimum inhibitory concentration (MIC) measurement was performed using broth microdilution assay. Nosocomial pathogens such as Enterococcus faecalis, Klebsiella pneumoniae, Bacillus subtilis, and Neisseria flavescens were isolated from mucormycosis. Veillonella parvula, Enterobacter hormaechei, and Acinetobacter calcoaceticus were isolated from chronic inflammatory lesion. Actinomyces massiliensis was isolated from actinomycosis in parotid gland. Capnocytophaga ochracea was isolated from actinomycosis in buccal region in anaerobic condition. There was no susceptible antibiotic to all bacteria in mucormycosis. Tetracycline was susceptible to all bacteria in chronic inflammation. C. ochracea was resistant to vancomycin and penicillin G; and other antibiotics showed susceptibility to all bacteria in actinomycosis. The results indicated that the combined treatment of two or more antibiotics is better than single antibiotic treatment in mucormycosis, and penicillin is the first recommended antibiotic to treat actinomycosis.

Characterization of antimicrobial resistance and application of RFLP for epidemiological monitoring of thermophilic Campylobacter spp. isolated from dogs and humans in Korea

  • Cho, Hyun-Ho;Kim, Sang-Hyun;Min, Wongi;Ku, Bok-Kyung;Kim, Jong-Hyun;Kim, Yong-Hwan
    • 대한수의학회지
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    • 제54권2호
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    • pp.91-99
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    • 2014
  • An antimicrobial susceptibility test was conducted to compare the resistance rates among Campylobacter spp. isolates from dogs (n = 50) raised under diverse conditions and humans (n = 50). More than 60% of Campylobacter (C.) jejuni from dogs and humans showed resistance to nalidixic acid, enrofloxacin and ciprofloxacin. C. jejuni isolates from humans showed higher resistance to tetracycline (83.3%) and ampicillin (91.3%) than those from dogs. None of the C. jejuni or Campylobacter coli isolates from humans or dogs were resistant to erythromycin. Overall, 85% of Campylobacter spp. isolates showed a multidrug resistant phenotype. Nucleotide sequencing analysis of the gryA gene showed that 100% of $NA^R/CIP^R$ C. jejuni isolates from dogs and humans had the Thr-$86^{th}$-Ile mutation, which is associated with fluoroquinolone resistance. flaA PCR restriction fragment length polymorphism (RFLP) typing to differentiate the isolates below the species level revealed 12 different clusters out of 73 strains. The human isolates belonged to eight different RFLP clusters, while five clusters contained dog and human isolates.

소아의 Transfusion Transmitted Virus-Like Minivirus 유병률 (Prevalence of Transfusion Transmitted Virus-Like Mini Virus in Children)

  • 정주영;한태희
    • Pediatric Infection and Vaccine
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    • 제11권2호
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    • pp.153-157
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    • 2004
  • 목 적 : TTV는 인체 감염이 확인된 최초의 circovirus로 간염을 유발할 가능성에 대해 연구가 이루어지고 있다. TLMV는 최근에 발견된 circovirus로 TTV보다 작지만 유사한 구조를 가진 것으로 알려져 있다. TLMV 감염의 성인 유병률은 약 70%인 것으로 알려지지만 소아의 유병률은 아직 확실하지 않다. 이에 저자들은 국내 소아의 TLMV 유병률을 알아보기 위하여 시행하였다. 방 법 : 2001년 6월부터 12월까지 인제의대 상계 백병원 외래를 방문한 환아중 TTV DNA에 대한 PCR이 시행되었던 88명의 혈청 검체를 대상으로 하였다. TLMV의 5'NCR(noncoding region) 특이적 시발체를 이용하여 PCR을 시행하였다. 1라운드 PCR은 M1359, M1365 시발체를 사용하여 $94^{\circ}C$ 10분, $94^{\circ}C$에서 40초, $60^{\circ}C$에서 40초, $72^{\circ}C$에서 50초, $72^{\circ}C$에서 10분의 조건에서 55회 시행하였다. 최종 산물 $2{\mu}L$에 M1360, M1366 시발체를 사용하여 1라운드와 동일한 조건에서 PCR을 55회 시행하였다. TLMV PCR 양성이 나온 10건에 대해 직접 염기 서열 분석과 계통 분석을 시행하였다. 결 과 : 소아 전체 연령에서 TLMV 감염 유병률은 49%였다. 연령별 유병률은 생후 1세 미만은 36%, 1~3세는 62%, 4~6세는 43%, 7~9세는 16%, 10~15세는 66%였다. 전체 소아의 22%에서 TTV와 TLMV의 혼합 감염이 확인되었다. TLMV PCR 산물 10건에 대한 염기 서열 분석을 시행한 결과 다른 나라의 TLMV 염기 서열과 많은 차이가 났다. 결 론 : 국내 소아의 TLMV 유병률은 49%로 비교적 높았으며 TLMV와 TTV와 혼합 감염이 발생함을 알 수 있었다. 국내에서 유행하는 TLMV의 유전형이 다른 나라와 큰 차이가 있을 가능성이 있지만 이에 대한 연구가 더 필요할 것으로 보인다.

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Opposite Roles of B7.1 and CD28 Costimulatory Molecules for Protective Immunity against HSV-2 Challenge in a gD DNA Vaccine Model

  • Weiner, David B.;Sin, Jeong-Im
    • IMMUNE NETWORK
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    • 제5권2호
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    • pp.68-77
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    • 2005
  • Background: Costimulation is a critical process in Ag-specific immune responses. Both B7.1 and CD28 molecules have been reported to stimulate T cell responses during antigen presentation. Therefore, we tested whether Ag-specific immune responses as well as protective immunity are influenced by coinjecting with B7.1 and CD28 cDNAs in a mouse HSV-2 challenge model system. Methods: ELISA was used to detect levels of antibodies, cytokines and chemokines while thymidine incorporation assay was used to evaluate T cell proliferation levels. Results: Ag-specific antibody responses were enhanced by CD28 coinjection but not by B7.1 coinjection. Furthermore, CD28 coinjection increased IgG1 production to a significant level, as compared to pgD+pcDNA3, suggesting that CD28 drives Th2 type responses. In contrast, B7.1 coinjection showed the opposite, suggesting a Th1 bias. B7.1 coinjection also enhanced Ag-specific Th cell proliferative responses as well as production of Th1 type cytokines and chemokines significantly higher than pgD+pcDNA3. However, CD28 coinjection decreased Ag-specific Th cell proliferative responses as well as production of Th1 types of cytokines and chemokine significantly lower than pgD+pcDNA3. Only MCP-1 production was enhanced by CD28. B7.1 coimmunized animals exhibited an enhanced survival rate as well as decreased herpetic lesion formation, as compared to pgD+pcDNA3. In contrast, CD28 vaccinated animals exhibited decreased survival from lethal challenge. Conclusion: This study shows that B7.1 enhances protective Th1 type cellular immunity against HSV-2 challenge while CD28 drives a more detrimental Th2 type immunity against HSV-2 challenge, supporting an opposite role of B7.1 and CD28 in Ag-specific immune responses to a Th1 vs Th2 type.

일본뇌염바이러스의 Mutant M 단백질에 반응하는 다클론항체의 생산: 극성 아미노산 잔기의 바이러스 생산과정에서의 역할 (Production of the Polyclonal Antibody That Recognizes the Mutant M Protein of Japanese Encephalitis Virus: Role of Its Charged Residues in Virus Production)

  • 김정민;윤상임;송병학;김진경;이영민
    • 미생물학회지
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    • 제46권2호
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    • pp.140-147
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    • 2010
  • 일본뇌염바이러스(Japanese encephalitis virus)는 모기 매개성 플라비바이러스에 속하며, 주로 동남아시아 지역에서 유행성 바이러스성 뇌염을 일으킨다. 일본뇌염바이러스는 외피를 가진 작은 바이러스로서, 양성가닥 RNA 게놈을 가지고 있다. 감염성을 띤 바이러스 입자는 capsid (C), membrane (M; prM 전구체로부터 생성), 및 envelope (E)과 같은 3개의 구조단백질로 이루어져 있다. 본 연구에서는 일본뇌염바이러스 생산과 정에 M 단백질의 N-말단부위에 위치한 극성 아미노산 잔기의 역할을 분석하였다. 일본뇌염바이러스의 infectious cDNA를 활용하여, M 단백질의 $E^9$$K^{15}K^{16}E^{17}$ 잔기를 알라닌으로 치환시킨 2개의 mutant cDNA (Mm1과 Mm2)를 각각 제작하였다. 각각의 cDNA로부터 합성된 mutant RNA를 세포에 트랜스펙션시킴으로써, 비록 세포 내에 축적된 3개의 구조단백질양은 변화가 없으나, 이들 세포로부터 생산된 바이러스의 양은 Mm2 RNA의 경우 ~1,000배 감소됨을 관찰하였다. 흥미롭게도, Mm2 RNA로부터 발현된 mutant M 단백질은 wild-type M 단백질을 인지하는 항혈청에는 반응하지 않았으나, mutant M 단백질을 항원으로 제작된 항혈청에는 반응하는 것을 알 수 있었다. 본 실험결과는 일본뇌염바이러스 M 단백질을 구성하는 3개의 극성 아미노산 잔기($K^{15}K^{16}E^{17}$)가 바이러스의 생산과정에 관여한다는 것을 암시한다. 앞으로, wild-type 또는 mutant M 단백질(Mm2)을 인식하는 2개의 항혈청은 이 단백질의 기능연구에 유용한 재료로 사용될 것으로 기대된다.

Species identification of Dyers woad leaf by DNA sequence of 5S-rRNA spacer domain and random amplified polymorphic DNA (RAPD) analysis

  • Zhao, K.J.;Dong, T.T.X.;Wong, Y.C.;Tu, P.F.;Tsim, K.W.K.
    • Advances in Traditional Medicine
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    • 제5권2호
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    • pp.117-123
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    • 2005
  • Dyers woad leaf (Daqingye) is a traditional Chinese medicine commonly used as anti-pyretic, anti-bacterial and anti-viral agent against infectious diseases. The Chinese Pharmacopoeia (2005) records that Dyers woad leaf should be derived from the leaves of Isatis indigotica Fort., but the leaves of Polygonum tinctorium Ait., Baphicacanthus cusia (Nees) Bremek. and Clerodendron cyrtophyllum Turcz. have also been used as substitutes of Dyers woad leaf in different regions of China. The leaf morphologies of these four species show a close resemblance, and based on their morphological appearance, it is difficult to identify them. Here, molecular genetic methods were developed as a target to identify different members of Dyers woad leaf. The 5S-rRNA spacer domain was amplified by polymerase chain reaction from genomic DNAs isolated from I. indigotica, P. tinctorium, B. cusia and C. cyrtophyllum, and the nucleotide sequences showed a great diversity. In addition, random amplification of polymorphic DNA analysis was also used to distinguish the members of Dyers woad leaf. These molecular methods could be used as a tool in authentic identification of Dyers woad leaf.

Comparative Analysis of the Complete Genome of Lactobacillus plantarum GB-LP2 and Potential Candidate Genes for Host Immune System Enhancement

  • Kwak, Woori;Kim, Kwondo;Lee, Chul;Lee, Chanho;Kang, Jungsun;Cho, Kyungjin;Yoon, Sook Hee;Kang, Dae-Kyung;Kim, Heebal;Heo, Jaeyoung;Cho, Seoae
    • Journal of Microbiology and Biotechnology
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    • 제26권4호
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    • pp.684-692
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    • 2016
  • Acute respiratory virus infectious diseases are a growing health problem, particularly among children and the elderly. Much effort has been made to develop probiotics that prevent influenza virus infections by enhancing innate immunity in the respiratory tract until vaccines are available. Lactobacillus plantarum GB-LP2, isolated from a traditional Korean fermented vegetable, has exhibited preventive effects on influenza virus infection in mice. To identify the molecular basis of this strain, we conducted a whole-genome assembly study. The single circular DNA chromosome of 3,284,304 bp was completely assembled and 3,250 protein-encoding genes were predicted. Evolutionarily accelerated genes related to the phenotypic trait of anti-infective activities for influenza virus were identified. These genes encode three integral membrane proteins, a teichoic acid export ATP-binding protein and a glucosamine - fructose-6-phosphate aminotransferase involved in host innate immunity, the nonspecific DNA-binding protein Dps, which protects bacteria from oxidative damage, and the response regulator of the three-component quorum-sensing regulatory system, which is related to the capacity of adhesion to the surface of the respiratory tract and competition with pathogens. This is the first study to identify the genetic backgrounds of the antiviral activity in L. plantarum strains. These findings provide insight into the anti-infective activities of L. plantarum and the development of preventive probiotics.