Visual screening for JAK2V617F mutation by a disposable dipstick

Abstract

During the last 5 years, it was discovered that the JAK2V617F somatic mutation is present in virtually all patients with polycythemia vera and a large proportion of patients with essential thrombocythemia, primary myelofibrosis, and refractory anemia with ring sideroblasts and thrombocytosis. As a result, JAK2V617F was incorporated as a new clonal marker in the 2008 revision of the WHO diagnostic criteria. Current methods for JAK2 genotyping include direct sequencing, pyrosequencing, allele-specific PCR with electrophoresis, restriction fragment length polymorphism, real-time PCR, DNA-melting curve analysis, and denaturing HPLC. Some of these methods are labor intensive and time consuming, while the others require specialized costly equipment and reagents. We report a method for direct detection of the JAK2V617F allele by the naked eye using a dipstick test in a dry-reagent format. The method comprises a triprimer PCR combined with visual detection of the products within minutes by the dipstick test. Specialized instrumentation is not involved. The requirements for highly qualified technical personnel are minimized. Because the detection reagents exist in dry form on the dipstick, there is no need for multiple pipetting and incubation steps.

Towards a Dual Lateral Flow Nanobiosensor for Simultaneous Detection of Virus Genotype-Specific PCR Products

“…The signal visualization was realized by Au NPs conjugated with anti-biotin antibodies. The anti-digoxigenin antibody performance for test zone construction was evaluated in the previously mentioned study [ 34 ], as well as other independent studies [ 37 , 38 ]. Therefore, the same type of anti-digoxigenin antibody was utilized for the TZ-S construction in the present study.…”

Paper-based nanobiosensors for diagnostics

“…The detection is done using streptavidin at a control line and an antibody specific for digoxin at a detection line. 49 A very interesting device was proposed by Kim et al, combining a microfluidic reverse transcription PCR (RT-PCR) reactor with a LFA to detect H1N1 virus (Fig. 3B).…”

Defining a molecular portrait of physical fitness

“…While this is appropriate for laboratory screening, it is not appropriate for point-of-care analysis. Ideally, mRNA, miRNA, and genetic analysis will need to be incorporated into a simple and easy-to-use device that requires minimal or no technical expertise, such as a microchip or dipstick [29, 30]. …”