TY - JOUR
T1 - Filmarray, an automated nested multiplex PCR system for multi-pathogen detection
T2 - Development and application to respiratory tract infection
AU - Poritz, Mark A.
AU - Blaschke, Anne J.
AU - Byington, Carrie L.
AU - Meyers, Lindsay
AU - Nilsson, Kody
AU - Jones, David E.
AU - Thatcher, Stephanie A.
AU - Robbins, Thomas
AU - Lingenfelter, Beth
AU - Amiott, Elizabeth
AU - Herbener, Amy
AU - Daly, Judy
AU - Dobrowolski, Steven F.
AU - Teng, David H.F.
AU - Ririe, Kirk M.
N1 - Copyright:
Copyright 2012 Elsevier B.V., All rights reserved.
PY - 2011
Y1 - 2011
N2 - The ideal clinical diagnostic system should deliver rapid, sensitive, specific and reproducible results while minimizing the requirements for specialized laboratory facilities and skilled technicians. We describe an integrated diagnostic platform, the "FilmArray", which fully automates the detection and identification of multiple organisms from a single sample in about one hour. An unprocessed biologic/clinical sample is subjected to nucleic acid purification, reverse transcription, a high-order nested multiplex polymerase chain reaction and amplicon melt curve analysis. Biochemical reactions are enclosed in a disposable pouch, minimizing the PCR contamination risk. FilmArray has the potential to detect greater than 100 different nucleic acid targets at one time. These features make the system well-suited for molecular detection of infectious agents. Validation of the FilmArray technology was achieved through development of a panel of assays capable of identifying 21 common viral and bacterial respiratory pathogens. Initial testing of the system using both cultured organisms and clinical nasal aspirates obtained from children demonstrated an analytical and clinical sensitivity and specificity comparable to existing diagnostic platforms. We demonstrate that automated identification of pathogens from their corresponding target amplicon(s) can be accomplished by analysis of the DNA melting curve of the amplicon.
AB - The ideal clinical diagnostic system should deliver rapid, sensitive, specific and reproducible results while minimizing the requirements for specialized laboratory facilities and skilled technicians. We describe an integrated diagnostic platform, the "FilmArray", which fully automates the detection and identification of multiple organisms from a single sample in about one hour. An unprocessed biologic/clinical sample is subjected to nucleic acid purification, reverse transcription, a high-order nested multiplex polymerase chain reaction and amplicon melt curve analysis. Biochemical reactions are enclosed in a disposable pouch, minimizing the PCR contamination risk. FilmArray has the potential to detect greater than 100 different nucleic acid targets at one time. These features make the system well-suited for molecular detection of infectious agents. Validation of the FilmArray technology was achieved through development of a panel of assays capable of identifying 21 common viral and bacterial respiratory pathogens. Initial testing of the system using both cultured organisms and clinical nasal aspirates obtained from children demonstrated an analytical and clinical sensitivity and specificity comparable to existing diagnostic platforms. We demonstrate that automated identification of pathogens from their corresponding target amplicon(s) can be accomplished by analysis of the DNA melting curve of the amplicon.
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U2 - 10.1371/journal.pone.0026047
DO - 10.1371/journal.pone.0026047
M3 - Article
C2 - 22039434
AN - SCOPUS:80054778674
VL - 6
JO - PLoS ONE
JF - PLoS ONE
SN - 1932-6203
IS - 10
M1 - e26047
ER -