Glycerol Reduces Cross Hybridization on Nitrocellulose Membrane

Fakultas Kedokteran Hewan, Universitas Gadjah Mada

2020-12

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Article

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Lateral flow assay (LFD) based nucleic acid lateral flow (NALF) method has been developed recently. The method met point of care testing (POCT) as a simple and rapid procedures, less equipment, and it can be performanced by less skilled technician. NALF based on nucleic acid hybridization is known more economical than immunochromatography assay which use antibody-antigen recognition. Cross-hybridization has issued while used to differentiate organism with high GC content and high homology as high similarity genome. Some techniques has applied some methods to give high stringency condition avoid cross hybridization reaction but need more procedure to apply. We found glycerol applied to buffer assay could reduce cross hybridization on nitrocellulose membrane. The study used 2 kinds of high stringency buffer there are PBS and SSC bases, , and high concentration of ssDNA amplicon as sample. Without glycerol ingredient gave cross hybridization signal on test line. But used glycerol could reduce those even omitted with PBS based buffer assay. Beside those, glycerol could significantly increased hybridization signal in SSC based buffer assay (p<0.05).