Both aerosol and primer dimer breakdown for straightforward genotyping based on an integrated immunochromatographic biosensor.

Straightforward genotyping can provide timely diagnostic information for diseases prevention and treatment. Taking advantages of speediness and convenience, although numerous genotyping strategies combined loop-mediated isothermal amplification (LAMP) and lateral flow have been reported to satisfy the demand of point-of-care test, the false positive result caused by aerosol and primer dimer as an innate conflict seriously limits their practical application. In this study, both aerosol and primer dimer as extrinsic and intrinsic inducements respectively are first broken through at one stroke based on an integrated immunochromatographic biosensor. By introducing digoxigenin labeled dUTP into LAMP, not only the amplicon can be analyzed through naked eye, but also the aerosol contamination can be eliminated thoroughly by uracil DNA glycosylase ignoring the open vessel. Primer dimer, the significant drawback in lateral flow-based strategies, has been overcome due to the bio-labeled deoxyribonucleotide and oligonucleotide cannot couple for signal generation even under the high primer concentration. Instead of colloidal gold, the gold magnetic nanoparticle is synthesized and assembled into this biosensor as a nanoprobe, which enables the result to be quantified by the magnetic signal for subjective bias elimination. The polymorphism of C677T in methylenetetrahydrofolate reductase, a crucial genetic code related to folate metabolism, is genotyped using saliva as noninvasive specimen dispense with DNA purification. Only 1 ng genomic DNA can provide accurate result within 25 min by a simple heater, which proves the potential of this biosensor to facilitate precision medicine.