title: Polymeric LabChip real-time PCR as a point-of-care-potential diagnostic tool for rapid detection of influenza A/H1N1 virus in human clinical specimens.
abstract: It is clinically important to be able to detect influenza A/H1N1 virus using a fast, portable, and accurate system that has high specificity and sensitivity. To achieve this goal, it is necessary to develop a highly specific primer set that recognizes only influenza A viral genes and a rapid real-time PCR system that can detect even a single copy of the viral gene. In this study, we developed and validated a novel fluidic chip-type real-time PCR (LabChip real-time PCR) system that is sensitive and specific for the detection of influenza A/H1N1, including the pandemic influenza strain A/H1N1 of 2009. This LabChip real-time PCR system has several remarkable features: (1) It allows rapid quantitative analysis, requiring only 15 min to perform 30 cycles of real-time PCR. (2) It is portable, with a weight of only 5.5 kg. (3) The reaction cost is low, since it uses disposable plastic chips. (4) Its high efficiency is equivalent to that of commercially available tube-type real-time PCR systems. The developed disposable LabChip is an economic, heat-transferable, light-transparent, and easy-to-fabricate polymeric chip compared to conventional silicon- or glass-based labchip. In addition, our LabChip has large surface-to-volume ratios in micro channels that are required for overcoming time consumed for temperature control during real-time PCR. The efficiency of the LabChip real-time PCR system was confirmed using novel primer sets specifically targeted to the hemagglutinin (HA) gene of influenza A/H1N1 and clinical specimens. Eighty-five human clinical swab samples were tested using the LabChip real-time PCR. The results demonstrated 100% sensitivity and specificity, showing 72 positive and 13 negative cases. These results were identical to those from a tube-type real-time PCR system. This indicates that the novel LabChip real-time PCR may be an ultra-fast, quantitative, point-of-care-potential diagnostic tool for influenza A/H1N1 with a high sensitivity and specificity.
authors.affiliation: Zoonosis Research Center, Department of Infection Biology, Wonkwang University School of Medicine, Iksan, Jeonbuk, Republic of Korea.
authors.forename: Hyun-Ok
authors.initials: HO
authors.lastname: Song
authors.forename: Je-Hyoung
authors.initials: JH
authors.lastname: Kim
authors.forename: Ho-Sun
authors.initials: HS
authors.lastname: Ryu
authors.forename: Dong-Hoon
authors.initials: DH
authors.lastname: Lee
authors.forename: Sun-Jin
authors.initials: SJ
authors.lastname: Kim
authors.forename: Deog-Joong
authors.initials: DJ
authors.lastname: Kim
authors.forename: In Bum
authors.initials: IB
authors.lastname: Suh
authors.forename: Du Young
authors.initials: du Y
authors.lastname: Choi
authors.forename: Kwang-Ho
authors.initials: KH
authors.lastname: In
authors.forename: Sung-Woo
authors.initials: SW
authors.lastname: Kim
authors.forename: Hyun
authors.initials: H
authors.lastname: Park
mesh_terms: D004526:Efficiency; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology / Q000821:virology; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods; D019095:Point-of-Care Systems; D011108:Polymers / Q000737:chemistry; D012367:RNA, Viral / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods; D012680:Sensitivity and Specificity; D013997:Time Factors; D054928:Validation Studies as Topic
chemical_list: D011108:Polymers; D012367:RNA, Viral