Pathogen lysis and nucleic acid extraction method using zinc oxide nanostar
The present invention relates to a method for lysing pathogen lysis and a method for extracting nucleic acid using zinc oxide nanostar, and the method for extracting nucleic acid using zinc oxide nanostar according to the present invention can extract nucleic acids by lysing cells of a pathogen without using a lysis buffer and can extract nucleic acid of high-purity and high-concentration by preventing nucleic acid degradation and fragmentation through various substances including salts contained in the lysis buffer at high concentration. In addition, the zinc oxide nanostar of the present invention (200 to 900 nm) has superior cell lysis capacity compared to the conventional zinc oxide nanoparticles (20 to 50 nm), thereby increasing the nucleic acid extraction efficiency and can extract at room temperature without a heating step to use as a field diagnostic method.
1. A method for extracting nucleic acid comprising:
a first step of preparing a mixture by adding zinc oxide nanostar to a nucleic acid sample and reacting;
a second step of preparing a reaction mixture by adding a diatomaceous earth modified with a silane compound to the mixture and adding at least one selected from the group consisting of dimethyl adipimidate (DMA), dimethyl pimelimidate (DMP), dimethyl suberimidate (DMS) and dimethyl 3,3′-dithiobispropionimidate (DTBP); and
a third step of extracting the nucleic acid from the reaction mixture,
wherein the zinc oxide nanostar has an average particle size of 200 to 600 nm and a plurality of pointed projections showed a particle shape arranged in a ring.
2. The method for extracting nucleic acid of claim 1 , wherein the silane compound is a compound represented by Chemical Formula 1:
wherein each of R 1 to R 3 may be same or different, and are any one of C1 to C4 alkyl or C1 to C4 alkoxy, and R 4 is anyone of amino(C1 to C10) alkyl, 3-(2-amino (C1 to C4)alkylamino) (C1 to C4)alkyl or 3-[2-(2-amino (C1 to C4)alkylamino) (C1 to C4) alkylamino] (C1 to C4)alkyl.
3. The method for extracting nucleic acid of claim 2 , wherein the silane compound is at least one selected from the group consisting of (3-aminopropyl) triethoxysilane (APTES), (3-aminopropyl)trimethoxysilane), (1-aminomethyl)triethoxysilane, (2-aminoethyl)triethoxysilane, (4-aminobutyl)triethoxysilane), (5-aminopentyl)triethoxysilane, (6-aminohexyl)triethoxysilane, 3-aminopropyl(diethoxy)methylsilane (APDMS), N-[3-(trimethoxysilyl)propyl]ethylenediamine, N-[3-(trimethoxysilyl)propyl]diethylenetriamine, [3-(2-aminoethylamino)propyl]trimethoxysilane (AEAPTMS) and 3-[(trimethoxysilyl)propyl]diethylenetriamine (TMPTA).