title: Evaluation of automated and manual DNA purification methods for detecting Ricinus communis DNA during ricin investigations.
abstract: In April of 2013, letters addressed to the President of United States and other government officials were intercepted and found to be contaminated with ricin, heightening awareness about the need to evaluate laboratory methods for detecting ricin. This study evaluated commercial DNA purification methods for isolating Ricinus communis DNA as measured by real-time polymerase chain reaction (PCR). Four commercially available DNA purification methods (two automated, MagNA Pure compact and MagNA Pure LC, and two manual, MasterPure complete DNA and RNA purification kit and QIAamp DNA blood mini kit) were evaluated. We compared their ability to purify detectable levels of R. communis DNA from four different sample types, including crude preparations of ricin that could be used for biological crimes or acts of bioterrorism. Castor beans, spiked swabs, and spiked powders were included to simulate sample types typically tested during criminal and public health investigations. Real-time PCR analysis indicated that the QIAamp kit resulted in the greatest sensitivity for ricin preparations; the MasterPure kit performed best with spiked powders. The four methods detected equivalent levels by real-time PCR when castor beans and spiked swabs were used. All four methods yielded DNA free of PCR inhibitors as determined by the use of a PCR inhibition control assay. This study demonstrated that DNA purification methods differ in their ability to purify R. communis DNA; therefore, the purification method used for a given sample type can influence the sensitivity of real-time PCR assays for R. communis.
authors: Bioterrorism and Emerging Pathogens Unit, North Carolina State Laboratory of Public Health, Raleigh, NC 27607, USA.
authors: Anne S
authors: AS
authors: Hutchins
authors: LPRB, DPEI, NCEZID, CDC, Atlanta, GA 30333, USA.
authors: Michael J
authors: MJ
authors: Astwood
authors: Bioterrorism and Emerging Pathogens Unit, North Carolina State Laboratory of Public Health, Raleigh, NC 27607, USA.
authors: J Royden
authors: JR
authors: Saah
authors: Bioterrorism Rapid Response and Advanced Technology (BRRAT) Laboratory, Laboratory Preparedness and Response Branch (LPRB), Division of Preparedness and Emerging Infections (DPEI), National Center for Emerging and Zoonotic Infectious Diseases (NCEZID), Centers for Disease Control and Prevention (CDC), Atlanta, GA 30333, USA.
authors: Pierre A
authors: PA
authors: Michel
authors: LPRB, DPEI, NCEZID, CDC, Atlanta, GA 30333, USA.
authors: Bruce R
authors: BR
authors: Newton
authors: Bioterrorism Rapid Response and Advanced Technology (BRRAT) Laboratory, Laboratory Preparedness and Response Branch (LPRB), Division of Preparedness and Emerging Infections (DPEI), National Center for Emerging and Zoonotic Infectious Diseases (NCEZID), Centers for Disease Control and Prevention (CDC), Atlanta, GA 30333, USA. Electronic address: Ldauphin@CDC.GOV.
authors: Leslie A
authors: LA
authors: Dauphin
mesh_terms: D057205:Automation, Laboratory; D002367:Ricinus communis / Q000737:chemistry; D002619:Chemical Warfare Agents / Q000032:analysis; D018744:DNA, Plant / Q000302:isolation & purification*; D010936:Plant Extracts / Q000235:genetics / Q000302:isolation & purification; D011208:Powders / Q000737:chemistry; D060888:Real-Time Polymerase Chain Reaction; D012276:Ricin / Q000032:analysis; D012278:Ricinus / Q000235:genetics*; D013048:Specimen Handling / Q000379:methods*
chemical_list: D002619:Chemical Warfare Agents; D018744:DNA, Plant; D010936:Plant Extracts; D011208:Powders; D012276:Ricin
keywords: Bioterrorism; Castor beans; DNA extraction; Ricin; Ricinus communis