• 제목/요약/키워드: polymerase

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Tuberculosis-associated hemophagocytic lymphohistiocytosis in adolescent diagnosed by polymerase chain reaction

  • Seo, Ju-Hee;Lee, Jun Ah;Kim, Dong Ho;Cho, Joongbum;Lim, Jung Sub
    • Clinical and Experimental Pediatrics
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    • 제59권1호
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    • pp.43-46
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    • 2016
  • We present a case of tuberculosis-associated hemophagocytic lymphohistiocytosis in a 14-year-old girl. The patient presented with weight loss, malaise, fatigue, prolonged fever, and generalized lymphadenopathy. Laboratory investigation revealed pancytopenia (white blood cells, $2,020cells/{\mu}L$; hemoglobin, 10.2 g/dL; platelets, $52,000cells/{\mu}L$), hypertriglyceridemia (229 mg/dL), and hyperferritinemia (1,420 ng/mL). Bone marrow biopsy showed a hypocellular bone marrow with a large numbers of histiocytes and marked hemophagocytosis; based on these findings, she was diagnosed with hemophagocytic lymphohistiocytosis. Polymerase chain reaction (PCR) with both the bone marrow aspiration and sputum samples revealed the presence of Mycobacterium tuberculosis. Antitubercular therapy with immune modulation therapy including dexamethasone and intravenous immunoglobulin was initiated. The results of all laboratory tests including bone marrow biopsy and PCR with both the bone marrow aspiration and sputum samples were normalized after treatment. Thus, early bone marrow biopsy and the use of techniques such as PCR can avoid delays in diagnosis and improve the survival rates of patients with tuberculosis-associated hemophagocytic lymphohistiocytosis.

MPN 및 H-NS 유전자를 표적으로 하는 PCR assay를 병용한 장염비브리오(Vibrio parahaemolyticus)의 정량 (Quantification of Vibrio parahaemolyticus Using a Most Probable Number-Polymerase Chain Reaction Assay Targeting the H-NS gene)

  • 김태옥;박권삼
    • 한국수산과학회지
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    • 제47권5호
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    • pp.556-561
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    • 2014
  • We applied a combination of most probable number-polymerase chain reaction (MPN-PCR) methods using a PCR procedure targeting the H-NS (VP1133) gene to detect Vibrio parahaemolyticus presence and density in seawater as well as within short-necked clam Ruditapes philippinarum tissues collected from Gomso Bay, Korea. In 30 seawater samples, V. parahaemolyticus levels ranged from less than 1.8 to $1.1{\times}10^3MPN/100mL$, and samples from August showed higher than those from other months. Furthermore, the levels of V. parahaemolyticus in six short-necked clam samples ranged from $7.8{\times}10^2$ to $2.1{\times}10^3MPN/100g$, approximately 2.5 times higher than in seawater samples from the corresponding month. Our results provide data on V. parahaemolyticus contamination in seawater and short-necked clam tissues, and help to improve quantitative methods of assessing V. parahaemolytcius levels.

Polymerase Chain Reaction 방법에 의한 Halobacteria gvp 유전자의 역전사 및 증폭 (Reverse Transcription and Amplification of Halobacterial gvp Genes with Polymerase Chain Reaction Method)

  • 윤병수;이상섭
    • 미생물학회지
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    • 제30권6호
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    • pp.456-459
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    • 1992
  • Halobacteria 의 gvpD. gvpE 유전자는 가스포 형성에 관여하는 유전자로, 이들은 그 transcripts 의 분석에 있어 특유의 연약성 때문에 많은 실험상의 문제를 야기시키고 있다. 본 실험은 연약한 mRNA 를 reverse transcriptase 를 사용, DNA 로 바꾸고 이를 다시 PCR(Polymerase Chain Reaction) 방법으로 증폭시킴으로써, 유전자의 연약한 mRNA 를 다시 상보적인 안정한 DNA 로 대치케 하여 RNA 상의 cloning, RNA sequencing 을 용이하게 하였다. 결과는 유전자 gvpD 에서 거의 전 ORF(Open Reading Frame) 의 범위에서 northern hybridization 에서 발견치 못한 transcipts 를 확인할 수 있었다.

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Detection of Lymphotropic Herpesviruses by Multiplex Polymerase Chain Reaction

  • Park, Sang-Tae;Kim, Seung-Han;Lee, Dong-Gun;Park, Jung-Hyun;Shin, Wan-Shik;Kim, Tai-Gyu;Paik, Soon-Young;Kim, Chun-Choo
    • Journal of Microbiology
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    • 제39권3호
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    • pp.226-228
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    • 2001
  • Human lymphotropic herpesvirus is known to be a major pathogen associated with various diseases in bone marrow transplantation (BMT) recipients. A multiplex nested-polymerase chain reaction (PCR) method was developed for the simultaneous detection of human lymphotropic herpesviruses, including Ebstein-Barr virus (EBV), cytomegalovirus (CMV), and human herpesvirus 6 variants A and B (HHV6-A, HHV6-B). To demonstrate the usefulness of multiplex PCR for the analysis of clinical samples, peripheral blood mononuclear cells and serum from BMT recipients were analysed. The results skewed that a clear detection could be made between EBV, HCMV and HHV-6. This multiplex PCR assay is an efficient and cost-effective approach to the analysis of large numbers of samples to determine the epidemiological importance of EBV HCMV and HHV-6.

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Changes in Poly ADP Ribose Polymerase Immune Response Cells of Cerebral Ischaemia Induced Rat by Transcranial Magnetic Stimulation of Alternating Current Approach

  • Koo, Hyun-Mo;Kim, Whi-Young
    • Journal of Magnetics
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    • 제19권4호
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    • pp.357-364
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    • 2014
  • This study examined effect of a transcranial magnetic stimulation device with a commercial-frequency approach on the neuronal cell death caused ischemia. For a simple transcranial magnetic stimulation device, the experiment was conducted on an ischemia induced rat by transcranial magnetic stimulation of a commercial-frequency approach, controlling the firing angle using a Triac power device. The transcranial magnetic stimulation device was controlled at a voltage of 220 V 60 Hz and the trigger of the Triac gate was varied from $45^{\circ}$ up to $135^{\circ}$. Cerebral ischemia was caused by ligating the common carotid artery of male SD rats and reperfusion was performed again to blood after 5 minutes. Protein Expression was examined by Western blotting and the immune response cells reacting to the antibodies of Poly ADP ribose polymerase in the cerebral nerve cells. As a result, for the immune response cells of Poly ADP ribose polymerase related to necrosis, the transcranial magnetic stimulation device suppressed necrosis and had a protective effect on nerve cells. The effect was greatest within 12 hours after ischemia. Therefore, it is believed that in the case of brain damage caused by ischemia, the function of brain cells can be restored and the impairment can be improved by the application of transcranial magnetic stimulation.

Molecular Modeling of Small Molecules as BVDV RNA-Dependent RNA Polymerase Allosteric Inhibitors

  • Chai, Han-Ha;Lim, Dajeong;Chai, Hee-Yeoul;Jung, Eunkyoung
    • Bulletin of the Korean Chemical Society
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    • 제34권3호
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    • pp.837-850
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    • 2013
  • Bovine viral diarrhea virus (BVDV), a major pathogen of cattle, is a well-characterized pestivirus which has been used as a good model virus for HCV. The RNA-dependent RNA polymerase (RdRp) plays a key role in the RNA replication process, thus it has been targeted for antivirus drugs. We employed two-dimensional quantitative structure-activity relationship (2D-QSAR) and molecular field analysis (MFA) to identify the molecular substructure requirements, and the particular characteristics resulted in increased inhibitory activity for the known series of compounds to act as effective BVDV inhibitors. The 2D-QSAR study provided the rationale concept for changes in the structure to have more potent analogs focused on the class of arylazoenamines, benzimidazoles, and acridine derivatives with an optimal subset of descriptors, which have significantly contributed to overall anti-BVDV activity. MFA represented the molecular patterns responsible for the actions of antiviral compound at their receptors. We conclude that the polarity and the polarizability of a molecule play a main role in the inhibitory activity of BVDV inhibitors in the QSAR modeling.

환경성 유해요인이 유전물질과 세포활성에 미치는 영향 V. CHO세포에서 세포주기에 따라 돌연변이원에 의해 유발된 DNA회복합성에 미치는 DNA중합효소의 역할 (Environmental Toxic Agents on Genetic Material and Cellular Ativity V. The Roles of DNA Polymerases on Mutagen-Induced DNA Repair Synthesis in Relation to Cell Cycle in Chinese Hamster Ovary Cells)

  • 엄경일;김춘광;신은주;문용석;이천복
    • 한국환경성돌연변이발암원학회지
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    • 제9권1호
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    • pp.23-32
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    • 1989
  • Chinese hamster ovary (CHO)-K1 cells echibited a differential sensitivity in the process of DNA repair synthesis induced by ethyl methanesulfonate (EMS) or bleomycin (BLM) in relation to cell cycle. Two assays were employed in this study: alkaline elution and unscheduled DNA synthesis. The post-treat-ment with aphidicolin (APC), an inhibitor of DNA polymerase alpha, inhibited DNA repair synthesis induced by EMS in G2 phase, while APC did not show any effect on BLM-induced DNA repair synthesis in all phases. On the other hands, the 2', 3'-dideoxythymidine (ddTTP), an inhibitor of DNA polymerase beta, inhibited DNA repair synthesis induced by EMS or BLM in both of G1 and G2 phases. These results suggested that the involvement of DNA polymerase alpha and beta in DNA repair was dependent on cell stage or used chemical agent.

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Application of digital polymerase chain reaction technology for noninvasive prenatal test

  • Lee, Seung Yong;Hwang, Seung Yong
    • Journal of Genetic Medicine
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    • 제12권2호
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    • pp.72-78
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    • 2015
  • Recently, noninvasive prenatal test (NIPT) has been adopted as a primary screening tool for fetal chromosomal aneuploidy. The principle of NIPT lies in isolating the fetal fraction of cell-free DNA in maternal plasma and analyzing it with bioinformatic tools to measure the amount of gene from the target chromosome, such as chromosomes 21, 18, and 13. NIPT will contribute to decreasing the need for unnecessary invasive procedures, including amniocentesis and chorionic villi sampling, for confirming fetal aneuploidy because of its higher positive predictive value than that of the conventional prenatal screening method. However, its greater cost than that of the current antenatal screening protocol may be an obstacle to the adoption of this innovative technique in clinical practice. Digital polymerase chain reaction (dPCR) is a novel approach for detecting and quantifying nucleic acid. dPCR provides real-time diagnostic advantages with higher sensitivity, accuracy, and absolute quantification than conventional quantitative PCR. Since the groundbreaking discovery that fetal cell-free nucleic acid exists in maternal plasma was reported, dPCR has been used for the quantification of fetal DNA and for screening for fetal aneuploidy. It has been suggested that dPCR will decrease the cost by targeting specific sequences in the target chromosome, and dPCR-based noninvasive testing will facilitate progress toward the implementation of a noninvasive approach for screening for trisomy 21, 18, and 13. In this review, we highlight the principle of dPCR and discuss its future implications in clinical practice.

Exon 8-9 Mutations of DNA Polymerase β in Ovarian Carcinoma Patients from Haldia, India

  • Khanra, Kalyani;Panda, Kakali;Mitra, A.K.;Sarkar, Ranu;Bhattacharya, Chandan;Bhattacharyya, Nandan
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권8호
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    • pp.4183-4186
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    • 2012
  • Background: Ovarian cancer is the number one killer among all the gynecological cancers. We undertook association study to identify potential alterations in the genomic DNA of a DNA repair gene, DNA polymerase beta ($pol{\beta}$), involved in base excision repair (BER), in ovarian carcinomas of patients from Haldia, India. Mutations, splice variants have been reported earlier in different tumors other than ovarian tumors. Aim: In this study we explored the possibility of association of any mutation of $pol{\beta}$ (Exon 8) with prognosis in 152 ovarian cancer samples. Results: Alteration in the exon 8 region (Exon 8:468, $A{\rightarrow}C$; 15.1%) was noted among fifty seven polymorphism positive samples. Alteration in the intervening sequence 8 (IVS8, -25, $A{\rightarrow}C$; 3.9%) was also noted. All alterations are heterozygous in nature. Conclusions: We found no significant association among the samples from serous type, stage IV, and the $pol{\beta}$ mutations ($P{\leq}0.01$). Only a slight tendency of association was evident between IVS8, -25, A to C; and stage III. Further analysis with a larger number of samples is needed.

Sonchus Yellow Net Virus에 감염된 Nicotiana edwardsonii 잎으로부터의 바이러스 단백질의 혈청학적 분석 (Serological Analysis of Sonchus Yellow Net Virus Proteins in Infected Nicotiana edwardsonii Leaf Tissues)

  • 최태진
    • 한국식물병리학회지
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    • 제14권3호
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    • pp.229-239
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    • 1998
  • Antibodies were raised against fusion proteins of the N-terminus and a region containing the GDNQ (Gly-Asp-Asn-Gln) polymerase motif of the L (polymerase) protein of sonchus yellow net virus (SYNV). Immunoblot analyses using these antibodies revealed the presence of the L protein in purified SYNV preparations and in nuclear extracts from infected tobacco. The serological analyses and detection in a polyacrylamide gels suggested that the L protein is present in at least a 20 fold lower abundance than the G, N, M1 and M2 proteins, and has size corresponding to a molecular weight of over 200 kDa as predicted from nucleotide sequence data. Electron microscopy with gold-labelled antibodies was used to localize the N, M2, and G proteins of SYNV in thin sections of infected tissue. When sections of SYNV-infected tissue were treated with antisera against total SYNV proteins and N protein, gold label could be detected in both the viroplasms and in virus particles. With the anti-M2 protein antiserum, the gold label was strongly localized in the viroplasms but only limited labelling of the virus particle sonly. Limited labelling of the L protein was observed in the viroplasms and the virus particles, presumably because of the low abundance of L protein in the tissues.

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