Magnetic Nanoparticles PCR Enzyme‐Linked Gene Assay for Quantitative Detection of BCR/ABL Fusion Gene in Chronic Myelogenous Leukemia

Abstract

This simple and cost-effective technique is suitable for monitoring CML patients during targeted therapy (tyrosine kinase inhibitors) especially in rural hospitals.

Solvent-sensitive nanoparticle-enhanced PCR assay for the detection of enterotoxigenic Escherichia coli

“…The optical density values were calculated into the relative value (ROD) according to Eq. ( 4 ) 7 . The number of LT-R-SSNPs that provided the highest ROD was the best for the SSNP-enhanced PCR assay and was used in further experiments.…”

“…Notably, the newly developed PCR-based technique, which was termed magnetic nanoparticles PCR enzyme-linked gene assay (MELGA), was introduced in the recent decade 6 – 9 . By applying forwards primer-modified MNPs to enrich the PCR product and biotinylated reverse primers to dramatically improve the sensitivity of postamplification analysis, MELGA could provide a limit of detection at the femtogram level 7 . According to a previous study, this technique exhibited 1,000 times higher sensitivity than that of conventional PCR 7 , 8 .…”

“…By applying forwards primer-modified MNPs to enrich the PCR product and biotinylated reverse primers to dramatically improve the sensitivity of postamplification analysis, MELGA could provide a limit of detection at the femtogram level 7 . According to a previous study, this technique exhibited 1,000 times higher sensitivity than that of conventional PCR 7 , 8 . Nevertheless, despite the high sensitivity, the signal developing step in MELGA postamplification was time-consuming due to multiple washing steps and prolonged reaction times between biotin-streptavidin and the enzyme–substrate.…”

“…For example, to increase the sensitivity of detection, magnetic nanoparticles (MNPs) with superparamagnetic properties and miscellaneous functionalization have been applied to isolate nucleic acids before amplification 2-5 . Notably, the newly developed PCR-based technique, which was termed magnetic nanoparticles PCR enzyme-linked gene assay (MELGA), was introduced in the recent decade [6][7][8][9] . By applying forwards primer-modified MNPs to enrich the PCR product and biotinylated reverse primers to dramatically improve the sensitivity of postamplification analysis, MELGA could provide a limit of detection at the femtogram level 7 .…”

Simultaneous detection of 45 fusion genes in leukemia by dual‐color fluorescence real‐time PCR

“…Consequently, this method is at risk of false‐positive results and often has low sensitivity . At the same time, many other platforms have been reported, including PCR enzyme‐linked gene assay, microfluidics‐based PCR, oligonucleotide microarray, and flow cytometric immunobead assay for detection of fusion proteins, but so far most of them are not widely used in clinical laboratories for many reasons .The recent high‐throughput methods such as next‐generation sequencing have complex procedures and are not yet sufficiently cost‐effective for routine testing. Therefore, an optimized method that would be easily applicable in clinical diagnostic laboratories for the simultaneous detection of large number of fusion genes is highly demanded.…”