title: Development and initial results of a low cost, disposable, point-of-care testing device for pathogen detection.
abstract: Development of small footprint, disposable, fast, and inexpensive devices for pathogen detection in the field and clinic would benefit human and veterinary medicine by allowing evidence-based responses to future out breaks. We designed and tested an integrated nucleic acid extraction and amplification device employing a loop-mediated isothermal amplification (LAMP) or reverse transcriptase-LAMP assay. Our system provides a screening tool with polymerase-chain-reaction-level sensitivity and specificity for outbreak detection, response, and recovery. Time to result is ∼90 min. The device utilizes a swab that collects sample and then transfers it to a disc of cellulose-based nucleic acid binding paper. The disc is positioned within a disposable containment tube with a manual loading port. In order to test for the presence of target pathogens, LAMP reagents are loaded through the tube's port into contact with the sample containing cellulose disc. The reagents then are isothermally heated to 63°C for ∼1 h to achieve sequence-specific target nucleic acid amplification. Due to the presence of a colorimetric dye, amplification induces visible color change in the reagents from purple to blue. As initial demonstrations, we detected methicillin resistant Staphylococcus aureus genomic DNA, as well as recombinant and live foot-and-mouth disease virus.
authors.affiliation: Lawrence Livermore National Laboratory, Livermore, CA 94550, USA. jane@llnl.gov
authors.forename: Jane P
authors.initials: JP
authors.lastname: Bearinger
authors.forename: Lawrence C
authors.initials: LC
authors.lastname: Dugan
authors.forename: Brian R
authors.initials: BR
authors.lastname: Baker
authors.forename: Sara B
authors.initials: SB
authors.lastname: Hall
authors.forename: Katja
authors.initials: K
authors.lastname: Ebert
authors.forename: Valerie
authors.initials: V
authors.lastname: Mioulet
authors.forename: Mikidache
authors.initials: M
authors.lastname: Madi
authors.forename: Donald P
authors.initials: DP
authors.lastname: King
mesh_terms: D003124:Colorimetry; D004269:DNA, Bacterial / Q000302:isolation & purification*; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics / Q000302:isolation & purification; D055624:Methicillin-Resistant Staphylococcus aureus / Q000235:genetics / Q000302:isolation & purification; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D019095:Point-of-Care Systems; D012367:RNA, Viral / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial; D012367:RNA, Viral