5 results on '"PT-PCR"'
Search Results
2. Identification of the Putative mRNA Coding for a Mitochondrial Isoform of Rat Ceruloplasmin.
- Author
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Vasin, A. V., Klotchenko, S. A., Platonova, N. A., Tsymbalenko, N. V., Babich, V. S., and Puchkova, L. V.
- Subjects
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GENE expression , *CERULOPLASMIN , *MESSENGER RNA , *GENES , *AMINO acids , *MITOCHONDRIA , *INTRONS - Abstract
A study was made of the expression of the ceruloplasmin (Cp) gene, whose product, blue multicopper ferroxidase, acts as a neuron survival factor. Computer analysis predicted an alternative promoter in the 3′-terminal region of intron 2 of the rat Cp gene and showed that transcription from this promoter potentially yields an mRNA coding for a 109-kDa polypeptide. The alternative Cp form starts with a region of 25 amino acid residues encoded by a part of intron 2. From Gly113, its sequence is identical to that of the major Cp form. The in silico data were confirmed experimentally by RT-PCR. The predicted mRNA was found predominantly in the liver and brain of adult rats. Direct sequencing of the PCR product demonstrated the complete identity of the predicted and real mRNA sequences. It was shown in vitro that a Cp-like protein of about 110 kDa is synthesized in the cytosol and accumulates in the absence of mitochondria. The protein was transferred into isolated mitochondria in a reconstructed system. Transport was energy-dependent, was associated with no change in Cp polypeptide length, and required cytosolic factors. Mitochondrial import of the Cp form was probably determined by an internal mitochondrial localization signal, KVVYREFTDSTFRE, corresponding to region 756–769 of mature Cp. The possible role of Cp in mitochondrial iron metabolism is discussed. [ABSTRACT FROM AUTHOR]
- Published
- 2005
- Full Text
- View/download PDF
3. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR
- Author
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Malik Peiris, Gabriel Goderski, Lisa Wijsman, Joanna Ellis, Jean Louis Romette, Olfert Landt, Chantal B.E.M. Reusken, Marie Luisa Schmidt, Bas van der Veer, Adam Meijer, Christian Drosten, Julia Schneider, Herman Goossens, Maria Zambon, Marco Kaiser, Richard Molenkamp, Marion Koopmans, Daniel K.W. Chu, Daphne G.J.C. Mulders, Tobias Bleicker, Bart L. Haagmans, Sharon van den Brink, Victor M. Corman, Sebastian Brünink, and Virology
- Subjects
0301 basic medicine ,Epidemiology ,Computer science ,medicine.disease_cause ,0302 clinical medicine ,Global health ,diagnostics ,030212 general & internal medicine ,COVID-19 ,RT-PCR ,Wuhan ,novel coronavirus ,2019-nCoV ,laboratory ,testing ,Coronavirus ,outbreak ,media_common ,Reverse Transcriptase Polymerase Chain Reaction ,European research ,3. Good health ,PT-PCR ,Severe acute respiratory syndrome-related coronavirus ,Coronavirus Infections ,medicine.medical_specialty ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,Pneumonia, Viral ,India ,03 medical and health sciences ,Betacoronavirus ,Virology ,medicine ,media_common.cataloged_instance ,European union ,Pandemics ,Clinical Laboratory Techniques ,SARS-CoV-2 ,Public health ,Research ,Public Health, Environmental and Occupational Health ,Outbreak ,Data science ,030104 developmental biology ,Workflow ,Coronavirus SARS-CoV-2 ,RNA ,Human medicine - Abstract
Background The ongoing outbreak of the recently emerged novel coronavirus (2019-nCoV) poses a challenge for public health laboratories as virus isolates are unavailable while there is growing evidence that the outbreak is more widespread than initially thought, and international spread through travellers does already occur. Aim We aimed to develop and deploy robust diagnostic methodology for use in public health laboratory settings without having virus material available. Methods Here we present a validated diagnostic workflow for 2019-nCoV, its design relying on close genetic relatedness of 2019-nCoV with SARS coronavirus, making use of synthetic nucleic acid technology. Results The workflow reliably detects 2019-nCoV, and further discriminates 2019-nCoV from SARS-CoV. Through coordination between academic and public laboratories, we confirmed assay exclusivity based on 297 original clinical specimens containing a full spectrum of human respiratory viruses. Control material is made available through European Virus Archive – Global (EVAg), a European Union infrastructure project. Conclusion The present study demonstrates the enormous response capacity achieved through coordination of academic and public laboratories in national and European research networks.
- Published
- 2020
- Full Text
- View/download PDF
4. Epidemiological Aspects of the Japanese Tobamovirus Strain, Pepper Mild Motte Virus(PMMoV) Infecting the L2 resistance Genotype of Green Pepper(Capsicum annuum L.)
- Author
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Toyoda, Kazuhiro, Hikichi, Yasufumi, Takeuchi, Shigeharu, Kuroda, Tomohisa, Okumura, Ako, Nasu, Yoshiko, Okuno, Tetsuro, and Suzuki, Kazumi
- Subjects
Pepper mild mottle virus(PMMoV) ,Capsicum annuum L ,fungi ,food and beverages ,resistance-breaking tobamovirus ,PT-PCR - Abstract
To understand the epidemiological aspects of tobamovirus infecting the L resistance genotypes of green pepper, fifteen isolates were collected from geographically different fields and were chracterized by their biological properties. All isolates infected L1 and L2 plants systemically, but were localized in L3 and L4 plants. The symptomatology on several test plants and the reactivity to an antiserum showed that they were identical to that of a Japanese strain of pepper mild mottle virus (PMMoV-J). The viral infection was also confirmed by a reverse transcription and polymerase chain reaction (RT-PCR) with oligonucleotide primers that amplity the coat protein gene of PMMoV-RNA. On the other hand, the RT-PCR allowed us to detect PMMoV in seeds of some commercial cultivars of green pepper. Viruses isolated from the seeds could infect L2 plants systemically. Further analysis of the nucleotide sequence of the predicted coat protein gene revealed that the isolates from the commercial seeds were identical to that of PMMoV-J. These results indicated that the L2 resistance-breaking tobamovirus has prevailed in fields of green pepper in Japan. and that infected seeds may be one of the initial sources of the viral infection.
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- 2004
5. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR
- Author
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Corman, Victor M., Landt, Olfert, Kaiser, Marco, Molenkamp, Richard, Meijer, Adam, Chu, Daniel K. W., Bleicker, Tobias, Brünink, Sebastian, Schneider, Julia, Schmidt, Marie Luisa, Mulders, Daphne G. J. C., Haagmans, Bart L., Van Der Veer, Bas, Van Den Brink, Sharon, Wijsman, Lisa, Goderski, Gabriel, Romette, Jean-Louis, Ellis, Joanna, Zambon, Maria, Peiris, Malik, Goossens, Herman, Reusken, Chantal, Koopmans, Marion P. G., and Drosten, Christian
- Subjects
Wuhan ,outbreak ,Coronavirus SARS-CoV-2 ,2019-nCoV ,diagnostics ,novel coronavirus ,COVID-19 ,laboratory ,testing ,600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit::610 Medizin und Gesundheit ,3. Good health ,PT-PCR - Abstract
Background: The ongoing outbreak of the recently emerged novel coronavirus (2019-nCoV) poses a challenge for public health laboratories as virus isolates are unavailable while there is growing evidence that the outbreak is more widespread than initially thought, and international spread through travellers does already occur. Aim: We aimed to develop and deploy robust diagnostic methodology for use in public health laboratory settings without having virus material available. Methods: Here we present a validated diagnostic workflow for 2019-nCoV, its design relying on close genetic relatedness of 2019-nCoV with SARS coronavirus, making use of synthetic nucleic acid technology. Results: The workflow reliably detects 2019-nCoV, and further discriminates 2019-nCoV from SARS-CoV. Through coordination between academic and public laboratories, we confirmed assay exclusivity based on 297 original clinical specimens containing a full spectrum of human respiratory viruses. Control material is made available through European Virus Archive - Global (EVAg), a European Union infrastructure project. Conclusion: The present study demonstrates the enormous response capacity achieved through coordination of academic and public laboratories in national and European research networks.
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