title: A pressure-driven column-based technique for the efficient extraction of DNA from respiratory samples.
abstract: Currently molecular techniques are a broadly accepted tool for diagnosis and are able to benefit patients in clinical practice. The polymerase chain reaction (PCR) has been especially incorporated into practical applications that are already in widespread use across the globe. With regard to the initial DNA extraction from clinically relevant samples, a number of commercially available kits are commonly used and are also designed to be easy to handle and less labor-intensive. In this study, the pressure system extracting DNA in column-based kit was developed, and its utility was compared with the centrifuge method using sputum from patients who were diagnosed with pneumonia. Also, due to the compact size and rapid processing time, the practical application of the pressure-based system incorporated into an automated pipetting machine was evaluated through clinical study. Our data suggests that DNA extraction by pressure was capable of serving as a substitute for the centrifuge method, and the compact and automatic nature of the pressure system device provided rapid and valuable information for clinical practice.
authors.affiliation: Department of Respiratory Medicine, Saitama Medical University, 38 Morohongou, Moroyama, Saitama 350-0495, Japan; Program in Cell Biology, Hospital for Sick Children, 686 Bay Street, Toronto, Ontario M5G 0A4, Canada. Electronic address: mycobacteriumtuberculosis@hotmail.com.
authors.forename: Takashi
authors.initials: T
authors.lastname: Hirama
authors.affiliation: Lifetech Co., Ltd, 4074 Miyadera, Iruma, Saitama, 358-0014, Japan.
authors.forename: Hajime
authors.initials: H
authors.lastname: Mogi
authors.affiliation: Department of Respiratory Medicine, Saitama Medical University, 38 Morohongou, Moroyama, Saitama 350-0495, Japan.
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Egashira
authors.affiliation: Lifetech Co., Ltd, 4074 Miyadera, Iruma, Saitama, 358-0014, Japan.
authors.forename: Eiji
authors.initials: E
authors.lastname: Yamamoto
authors.affiliation: Lifetech Co., Ltd, 4074 Miyadera, Iruma, Saitama, 358-0014, Japan.
authors.forename: Shigenari
authors.initials: S
authors.lastname: Kukisaki
authors.affiliation: Department of Respiratory Medicine, Saitama Medical University, 38 Morohongou, Moroyama, Saitama 350-0495, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Hagiwara
authors.affiliation: Lifetech Co., Ltd, 4074 Miyadera, Iruma, Saitama, 358-0014, Japan.
authors.forename: Osamu
authors.initials: O
authors.lastname: Takei
mesh_terms: D002498:Centrifugation; D004269:DNA, Bacterial / Q000302:isolation & purification*; D006193:Haemophilus influenzae / Q000737:chemistry / Q000254:growth & development / Q000472:pathogenicity; D006801:Humans; D007711:Klebsiella pneumoniae / Q000737:chemistry / Q000254:growth & development / Q000472:pathogenicity; D001936:Moraxella catarrhalis / Q000737:chemistry / Q000254:growth & development / Q000472:pathogenicity; D018410:Pneumonia, Bacterial / Q000175:diagnosis* / Q000382:microbiology; D011312:Pressure; D011550:Pseudomonas aeruginosa / Q000737:chemistry / Q000254:growth & development / Q000472:pathogenicity; D011933:Reagent Kits, Diagnostic; D060888:Real-Time Polymerase Chain Reaction; D052616:Solid Phase Extraction / Q000295:instrumentation / Q000379:methods*; D013183:Sputum / Q000737:chemistry*; D013296:Streptococcus pneumoniae / Q000737:chemistry / Q000254:growth & development / Q000472:pathogenicity
chemical_list: D004269:DNA, Bacterial; D011933:Reagent Kits, Diagnostic
keywords: Column; DNA extraction; HIRA-TAN; Pressure; Real-time PCR