| Rapid non-enzymatic extraction method for isolating PCR-quality camelpox virus DNA from skin. | |
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MedLine Citation:
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PMID: 20654652 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Molecular diagnostic investigations of orthopoxvirus (OPV) infections are performed using a variety of clinical samples including skin lesions, tissues from internal organs, blood and secretions. Skin samples are particularly convenient for rapid diagnosis and molecular epidemiological investigations of camelpox virus (CMLV). Classical extraction procedures and commercial spin-column-based kits are time consuming, relatively expensive, and require multiple extraction and purification steps in addition to proteinase K digestion. A rapid non-enzymatic procedure for extracting CMLV DNA from dried scabs or pox lesions was developed to overcome some of the limitations of the available DNA extraction techniques. The procedure requires as little as 10mg of tissue and produces highly purified DNA [OD(260)/OD(280) ratios between 1.47 and 1.79] with concentrations ranging from 6.5 to 16 microg/ml. The extracted CMLV DNA was proven suitable for virus-specific qualitative and, semi-quantitative PCR applications. Compared to spin-column and conventional viral DNA extraction techniques, the two-step extraction procedure saves money and time, and retains the potential for automation without compromising CMLV PCR sensitivity. |
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Authors:
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A Ausama Yousif; A Abdelmohsen Al-Naeem; M Ahmad Al-Ali |
Publication Detail:
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Type: Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't Date: 2010-07-21 |
Journal Detail:
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Title: Journal of virological methods Volume: 169 ISSN: 1879-0984 ISO Abbreviation: J. Virol. Methods Publication Date: 2010 Oct |
Date Detail:
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Created Date: 2010-08-30 Completed Date: 2010-12-08 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 8005839 Medline TA: J Virol Methods Country: Netherlands |
Other Details:
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Languages: eng Pagination: 138-42 Citation Subset: IM |
Copyright Information:
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Copyright (c) 2010 Elsevier B.V. All rights reserved. |
Affiliation:
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Central Biotechnology Laboratory, College of Veterinary Medicine and Animal Resources, Bldg. 999, King Faisal University, Al-Hufof, 31982, Al-Ahsaa, Saudi Arabia. ausama_yousif@yahoo.com |
| Data Bank Information | |
Bank Name/Acc. No.:
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GENBANK/GU937751 |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Camels DNA, Viral / isolation & purification* Molecular Sequence Data Orthopoxvirus / isolation & purification* Polymerase Chain Reaction / methods Poxviridae Infections / veterinary*, virology Sensitivity and Specificity Sequence Analysis, DNA Skin / virology* Virology / methods* |
| Chemical | |
Reg. No./Substance:
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0/DNA, Viral |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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