Porous solid phase for rapidly isolating biological molecules for nucleic acid amplification reaction from biological sample, and use thereof

Abstract

The present invention provides a method for rapidly ascertaining whether or not a target sequence exists in a biological sample by: rapidly isolating biological samples for a nucleic acid amplification reaction from the biological sample by using a porous solid phase capable of rapidly sucking biological samples present in a biological sample into pores; and amplifying the same.

1 . A method for rapidly isolating a biological molecule for a nucleic acid amplification reaction from a biological sample, the method comprising putting a porous solid phase in contact with a biological sample to get the biological molecule present in the biological sample sucked into the pores of the porous solid phase. 
     
     
         2 . The method of  claim 1 , wherein the biological sample is derived from an animal, a plant, a bacterium, or a fungus. 
     
     
         3 . The method of  claim 1 , wherein the biological molecule is DNA, RNA, dsRNA, microRNA, viroid, virus, bacteria, fungi or microalgae. 
     
     
         4 . The method of  claim 1 , wherein the nucleic acid amplification reaction is cDNA synthesis, polymerase chain reaction (PCR), multiplex PCR or reverse transcriptase polymerase chain reaction (RT-PCR). 
     
     
         5 . The method of  claim 1 , wherein the porous solid phase is at least one selected from the group consisting of carbonized cellulose, paper held together in a particle form, natural or synthetic zeolite, polystylene, polycarbonate, polyprophylene, porous metal particles, porous rubber, microporous glass held together in a particle form, lime, a shell, a ceramic fragment, and ceramic of oxide material. 
     
     
         6 . The method of  claim 5 , wherein a major component of the ceramic of oxide material is Al 2 O 3 , Fe 2 O 3 , low temperature co-fired ceramic (LTCC), PbO or ZnO. 
     
     
         7 . The method of  claim 1 , wherein the porous solid phase is in the shape of a cube, a cuboid, a sphere, a cylinder, a bar, a bar with a dent at one end or a bar with a sharp end. 
     
     
         8 . A method for amplifying a target sequence in a biological sample, the method comprising:
 (a) putting a porous solid phase in contact with a biological sample, in which biological molecules of interest are sucked into the pores of the porous solid phase; and   (b) adding the porous solid phase in step (a), into which the biological molecule are sucked, as a template for a nucleic acid amplification reaction, and then performing the amplification reaction using a target primer set to amplify the target sequence.   
     
     
         9 . The method of  claim 8 , wherein the nucleic acid amplification reaction is a polymerase chain reaction (PCR). 
     
     
         10 . A method for amplifying a target sequence in a biological sample, the method comprising:
 (a) putting a porous solid phase in contact with a biological sample, in which biological molecules of interestare sucked into the pores of the porous solid phase; and   (b) adding reverse transcriptase to the porous solid phase in step (a), into which the biological molecules are sucked, and performing a reverse transcriptase reaction; and   (c) adding the reverse transcriptase reaction product as a template for a nucleic acid amplification reaction, and performing an amplification reaction using a target primer set to amplify the target sequence.   
     
     
         11 . A method for rapidly detecting the presence of a target sequence in a biological sample, the method comprising:
 (a) putting a porous solid phase in contact with a biological sample, in which biological molecules of interest into the pores of the porous solid phase;   (b) adding the porous solid phase in step (a), into which the biological molecule are sucked, as a template for a nucleic acid amplification reaction, and performing an amplification reaction using a target primer set to amplify the target sequence; and   (c) detecting the amplified product.   
     
     
         12 . A method for rapidly detecting the presence of a target sequence in a biological sample, the method comprising:
 (a) putting a porous solid phase in contact with a biological sample, in which biological molecules of interest into the pores of the porous solid phase;   (b) adding reverse transcriptase to the porous solid phase in step (a), into which the biological molecule are sucked, and performing a reverse transcriptase reaction;   (c) adding the reverse transcriptase reaction product as a template for a nucleic acid amplification reaction, and performing an amplification reaction using a target primer set to amplify the target sequence; and   (d) detecting the amplified product.   
     
     
         13 . A kit for a nucleic acid amplification reaction for amplifying a target sequence in a biological sample, the kit comprising a porous solid phase capable of rapidly sucking a biological molecule present in the biological sample into the pores of the porous solid phase; a target primer set; and a reagent for performing an amplification reaction. 
     
     
         14 . The kit of  claim 13 , wherein the reagent for performing an amplification reaction is a reverse transcriptase, a DNA polymerase, dNTPs and a buffer solution. 
     
     
         15 . The kit of  claim 13 , wherein the porous solid phase is at least one selected from the group consisting of carbonized cellulose, paper held together in a particle form, natural or synthetic zeolite, polystylene, polycarbonate, polyprophylene, porous metal particles, porous rubber, microporous glass held together in a particle form, lime, a shell, a ceramic fragment, and ceramic of oxide material. 
     
     
         16 . A kit for rapidly isolating a biological molecule for a nucleic acid amplification reaction from a biological sample, the kit comprising a porous solid phase capable of rapidly sucking the biological molecule present in the biological sample into the pores of the porous solid phase. 
     
     
         17 . A composition for rapidly isolating a biological molecule for a nucleic acid amplification reaction from a biological sample, the composition comprising a porous solid phase capable of rapidly sucking the biological molecule present in the biological sample into the pores of the porous solid phase. 
     
     
         18 . The composition of  claim 17 , wherein the porous solid phase is at least one selected from the group consisting of carbonized cellulose, paper held together in a particle form, natural or synthetic zeolite, polystylene, polycarbonate, polyprophylene, porous metal particles, porous rubber, microporous glass held together in a particle form, lime, a shell, a ceramic fragment, and ceramic of oxide material.