title: Integrated DNA and RNA extraction and purification on an automated microfluidic cassette from bacterial and viral pathogens causing community-acquired lower respiratory tract infections.
abstract: In this paper, we describe the development of an automated sample preparation procedure for etiological agents of community-acquired lower respiratory tract infections (CA-LRTI). The consecutive assay steps, including sample re-suspension, pre-treatment, lysis, nucleic acid purification, and concentration, were integrated into a microfluidic lab-on-a-chip (LOC) cassette that is operated hands-free by a demonstrator setup, providing fluidic and valve actuation. The performance of the assay was evaluated on viral and Gram-positive and Gram-negative bacterial broth cultures previously sampled using a nasopharyngeal swab. Sample preparation on the microfluidic cassette resulted in higher or similar concentrations of pure bacterial DNA or viral RNA compared to manual benchtop experiments. The miniaturization and integration of the complete sample preparation procedure, to extract purified nucleic acids from real samples of CA-LRTI pathogens to, and above, lab quality and efficiency, represent important steps towards its application in a point-of-care test (POCT) for rapid diagnosis of CA-LRTI.
authors.affiliation: Department of Medical Microbiology, Vaccine & Infectious Disease Institute (VAXINFECTIO), Universiteit Antwerpen, Antwerp, Belgium.
authors.forename: Liesbet
authors.initials: L
authors.lastname: Van Heirstraeten
authors.forename: Peter
authors.initials: P
authors.lastname: Spang
authors.forename: Carmen
authors.initials: C
authors.lastname: Schwind
authors.forename: Klaus S
authors.initials: KS
authors.lastname: Drese
authors.forename: Marion
authors.initials: M
authors.lastname: Ritzi-Lehnert
authors.forename: Benjamin
authors.initials: B
authors.lastname: Nieto
authors.forename: Marta
authors.initials: M
authors.lastname: Camps
authors.forename: Bryan
authors.initials: B
authors.lastname: Landgraf
authors.forename: Francesc
authors.initials: F
authors.lastname: Guasch
authors.forename: Antoni Homs
authors.initials: AH
authors.lastname: Corbera
authors.forename: Josep
authors.initials: J
authors.lastname: Samitier
authors.forename: Herman
authors.initials: H
authors.lastname: Goossens
authors.forename: Surbhi
authors.initials: S
authors.lastname: Malhotra-Kumar
authors.forename: Tina
authors.initials: T
authors.lastname: Roeser
mesh_terms: D053000:Analytic Sample Preparation Methods; D001331:Automation; D001419:Bacteria / Q000235:genetics; D018407:Bacterial Physiological Phenomena; D017714:Community-Acquired Infections / Q000382:microbiology* / Q000821:virology*; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification*; D006801:Humans; D009980:Influenza A virus / Q000235:genetics / Q000502:physiology; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification*; D012141:Respiratory Tract Infections / Q000382:microbiology* / Q000821:virology*
chemical_list: D004269:DNA, Bacterial; D012367:RNA, Viral