Enclosed unit for rapid detection of a target nucleic acid amplification product

Abstract

The invention relates to a method for rapid detection of a target nucleic acid amplification product while preventing cross-contamination between target nucleic acid amplification products and avoiding false positives, comprising the steps of: a) leaving the reaction tube unopened after the amplification reaction is finished, so as to prevent the target nucleic acid amplification product from leaking out and resulting in contamination; b) placing the unopened reaction tube inside an enclosed unit, making the target nucleic acid amplification product be transferred to a test strip from the reaction tube in a physically enclosed environment; c) performing detection in a visual read-out manner, and determining the result; d) discarding the enclosed unit in a safety place as a whole without opening it after the detection. The invention also relates to a totally enclosed unit for detecting a target nucleic acid amplification product, and still relates to applications of the totally enclosed rapid detection unit in detection of infectious pathogens, food industry, agriculture, livestock husbandry, customs quarantine control, and determination of DNA.

The invention claimed is: 
     
         1 .- 11 . (canceled) 
     
     
         12 . A method of detecting a target nucleic acid amplification product which is retained in a detection device thereby avoiding contamination of a proximate space comprising the steps of:
 a) placing the sealed container containing a target nucleic acid amplification product into the detection device;   b) closing the detection device such that the sealed container is punctured and the fluid containing the target nucleic amplification product is transferred to a sterile visual detection strip within the device such that the fluid is retained within the detection device; and   c) detecting the target nucleic acid amplification product.   
     
     
         13 . The method of  claim 12 , wherein the detection device cannot be reopened once it is closed thereby preventing the fluid from leaking out of the detection device. 
     
     
         14 . A method of detecting a target nucleic acid amplification product which is retained in a detection device thereby avoiding contamination of a proximate space comprising the steps of:
 a) placing the sealed container containing a target nucleic acid amplification product into the detection device which comprises a washing buffer tube containing washing buffer;   b) closing the detection device such that the sealed container is punctured and the washing buffer tube is punctured and the fluid containing the target nucleic amplification product is combined with the washing buffer to form a mixture of target nucleic acid amplification product and washing buffer such that the mixture is transferred to a sterile visual detection strip within the device such that the fluid is retained within the detection device; and   c) detecting the target nucleic acid amplification product.   
     
     
         15 . The method of  claim 14 , wherein the detection device cannot be reopened once it is closed thereby preventing the fluid from leaking out of the detection device. 
     
     
         16 . The method of  claim 12  wherein the detection device comprises a cassette having a port into which a container containing the target nucleic acid amplification product in a fluid is placed such that a portion of the container is exposed, a sealable outer casing into which the cassette is placed and which completely encompasses the cassette when the outer casing is sealed, a cutting device that punctures the exposed portion of the container when the sealable outer casing is sealed to release the fluid and a detection strip in proximity to the container such that the liquid contacts the detection strip when the tube is punctured. 
     
     
         17 . The method of  claim 12  wherein the detection device comprises an inner core ( 1 ) and an outer casing ( 15 ), wherein the inner core ( 1 ) comprises a fixing case ( 2 ) and a base ( 5 ), the fixing case ( 2 ) having two ports respectively for housing washing buffer vacuoles ( 3 ) and a reaction tube ( 11 ) containing an amplification product ( 7 ), the base ( 5 ) comprising a washing buffer container ( 21 ) having a vacuole puncture needle ( 4 ), an amplification product container ( 22 ) having a blade ( 9 ), a sealing diaphragm ( 6 ), a piece of glassfiber paper ( 8 ), and a test strip sealing part having a test strip ( 13 ) sealed therein and a transparent window ( 12 ) and wherein the outer casing ( 15 ) comprises a handle cover ( 16 ), a fixing case pressing part ( 17 ), a washing buffer vacuole extruding part ( 18 ), and a transparent window ( 20 ). 
     
     
         18 . The method of  claim 12  wherein the detection device comprises an inner core ( 1 ) and an outer casing ( 15 ), wherein the inner core ( 1 ) comprises a fixing case ( 2 ) and a base ( 5 ), the fixing case ( 2 )) having two ports respectively for housing washing buffer vacuoles ( 3 ) and a reaction tube ( 11 ) containing an amplification product ( 7 ); the base part ( 5 ) comprising a washing buffer container ( 21 ) having a vacuole puncture needle ( 4 ) for puncturing the washing buffer vacuoles ( 3 ), an amplification product container ( 22 ) having a blade ( 9 ) for cutting the reaction tube ( 11 ), a sealing diaphragm ( 6 ) between the elements ( 21 ) and ( 22 ), a piece of glassfiber paper ( 8 ) disposed in the container elements ( 21 ) and ( 22 ) and connected to the bottom of a test strip ( 13 ); and a test strip sealing part having the test strip ( 13 ) sealed therein and a transparent window ( 12 ) for observing the test strip ( 13 ), and the outer casing ( 15 ) comprises a handle cover ( 16 ), a fixing case pressing part ( 17 ), a washing buffer vacuole extruding part ( 18 ), and a transparent window ( 20 ) in parallel with the transparent window ( 12 ). 
     
     
         19 . The method of  claim 17  wherein the detection device further comprises a sealing ring ( 10 ) positioned between the hole for housing the reaction tube ( 11 ) and the base ( 5 ), such that the amplification product will not flow back into the fixing case ( 2 ). 
     
     
         20 . The method of  claim 18  wherein the detection device further comprises a sealing ring ( 10 ) positioned between the hole for housing the reaction tube ( 11 ) and the base ( 5 ), such that the amplification product will not flow back into the fixing case ( 2 ). 
     
     
         21 . The method of  claim 19  wherein the detection device further comprises further comprising a sealing diaphragm ( 6 ) comprising three alternating layers vertically disposed such that the amplification product will not flow back into the fixing case ( 2 ), while being mixed with the washing buffer flowing in a single direction and absorbed by the test strip ( 13 ). 
     
     
         22 . The method of  claim 20  wherein the detection device further comprises further comprising a sealing diaphragm ( 6 ) comprising three alternating layers vertically disposed such that the amplification product will not flow back into the fixing case ( 2 ), while being mixed with the washing buffer flowing in a single direction and absorbed by the test strip ( 13 ). 
     
     
         23 . The method of  claim 12  wherein the amplification product is DNA. 
     
     
         24 . The method of  claim 23  wherein the DNA is from a pathogenic organism. 
     
     
         25 . The method of  claim 23  wherein the DNA has a genetic mutation. 
     
     
         26 . A method of detecting a target nucleic acid amplification product which prevents contamination of the target nucleic acid amplification product by contamination product(s) comprising the steps of:
 a) placing the sealed container containing a target nucleic acid amplification product into the detection device;   b) closing the device such that the sterile sealed container is punctured and the fluid containing the target nucleic amplification product is transferred to a sterile detection strip within the device such that no contamination products leak to the environment, and such that cross-contaminations between testing samples are prevented, and such that the risks of false positive results are reduced; and   c) detecting the target nucleic acid amplification product.