Rapid non-enzymatic extraction method for isolating PCR-quality camelpox virus DNA from skin

Journal of Virological Methods
A Ausama YousifM Ahmad Al-Ali

Abstract

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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Citations

Jan 16, 2014·Indian Journal of Virology : an Official Organ of Indian Virological Society·Vinayagamurthy BalamuruganRaj Kumar Singh
Jun 24, 2014·Virus Genes·Ramasamy SanthamaniMuthannan Andavar Ramakrishnan
Jul 11, 2014·Archives of Virology·M MosadeghhesariH R Varshovi
Sep 29, 2011·Antiviral Research·Sophie DuraffourRobert Snoeck
Jan 6, 2017·Vector Borne and Zoonotic Diseases·Abdelmalik I Khalafalla, Fatima Abdelazim

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