title: Toward point-of-care testing for JAK2 V617F mutation on a microchip.
abstract: Molecular genetics now plays a crucial role in diagnosis, the identification of prognostic markers, and monitoring of hematological malignancies. Demonstration of acquired changes such as the JAK2 V617F mutation within myeloproliferative neoplasms (MPN) has quickly moved from a research setting to the diagnostic laboratory. Microfluidics-based assays can reduce the assay time and sample/reagent consumption and enhance the reaction efficiency; however, no current assay has integrated isothermal amplification for point-of-care MPN JAK2 V617F mutation testing with a microchip. In this report, an integrated microchip that performs the whole human blood genomic DNA extraction, loop-mediated isothermal nucleic acid amplification (LAMP) and visual detection for point-of-care genetic mutation testing is demonstrated. This method was validated on DNA from cell lines as well as on whole blood from patients with MPN. The results were compared with those obtained by unlabeled probe melting curve analysis. This chip enjoys a high accuracy, operability, and cost/time efficiency within 1h. All these benefits provide the chip with a potency toward a point-of-care genetic analysis. All samples identified as positive by unlabeled probe melting curve analysis (n=27) proved positive when tested by microchip assay. None of the 30 negative controls gave false positive results. In addition, a patient with polycythemia vera diagnosed as being JAK2 V617F-negative by unlabeled probe melting curve analysis was found to be positive by the microchip. This microchip would possibly be very attractive in developing a point-of-care platform for quick preliminary diagnosis of MPN or other severe illness in resource-limited settings.
authors.affiliation: Department of Laboratory Medicine, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors.forename: Hua
authors.initials: H
authors.lastname: Wang
authors.affiliation: Department of Laboratory Medicine, Shanghai Tenth People's Hospital, Tongji University, Shanghai 200072, China.
authors.forename: Weiwei
authors.initials: W
authors.lastname: Liu
authors.affiliation: Center of Laboratory, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors.forename: Xinju
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Center of Laboratory, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Xu
authors.affiliation: Department of Laboratory Medicine, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China; Center of Laboratory, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Kang
authors.affiliation: Department of Laboratory Medicine, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China; Center of Laboratory, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors.forename: Shibao
authors.initials: S
authors.lastname: Li
authors.affiliation: Department of Laboratory Medicine, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China; Center of Laboratory, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors.forename: Zhiyuan
authors.initials: Z
authors.lastname: Wu
authors.affiliation: Institute of Microanalytical Systems, Department of Chemistry, Zhejiang University, Hangzhou 310058, China.
authors.forename: Zhiliu
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Institute of Microanalytical Systems, Department of Chemistry, Zhejiang University, Hangzhou 310058, China. Electronic address: yaobo08@zju.edu.cn.
authors.forename: Bo
authors.initials: B
authors.lastname: Yao
authors.affiliation: Department of Laboratory Medicine, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China; Center of Laboratory, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China. Electronic address: guanming88@yahoo.com.
authors.forename: Ming
authors.initials: M
authors.lastname: Guan
mesh_terms: D016022:Case-Control Studies; D006801:Humans; D053614:Janus Kinase 2 / Q000097:blood / Q000235:genetics*; D009154:Mutation; D009196:Myeloproliferative Disorders / Q000097:blood / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D000067716:Point-of-Care Testing; D011087:Polycythemia Vera / Q000175:diagnosis / Q000235:genetics
chemical_list: D053614:Janus Kinase 2
keywords: JAK2 V671F mutation; Microchip; Myeloproliferative neoplasms; Point-of-care testing