Reaction vessel for PCR device and method of performing PCR
Inventors
Lüdke, Gerd • Boos, Andreas • Motejadded, Hassan • Bacher, Johannes
Assignees
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Publication Number
US-9592511-B2
Publication Date
2017-03-14
Expiration Date
Abstract
The present invention provides a reaction vessel (20) for a PCR device. The reaction vessel (20) comprises a sample vial (32) defining a reaction chamber (33) for performing PCR and a storage vessel (62) defining a storage chamber (63) for optical detection. The reaction chamber (33) is in fluid communication with a liquid supply port (34) for supplying a liquid sample containing at least one target DNA to the reaction chamber (33). The reaction chamber (33) and the storage chamber (63) are in fluid communication via a spacer element (42) and a porous membrane (51) for hybridization of the at least one target DNA within the liquid sample onto specific immobilised hybridization probes. The lower end of the spacer element (42) extends into the reaction chamber (33), but does not reach the bottom thereof. The upper end of the spacer element (42) is located in proximity of the porous membrane (51), which is made from a material having different physical properties in a dry state and a wet state. In the dry state the porous membrane (51) allows air as well as liquid to pass therethrough. In the wet state the porous membrane (51) still allows the passage of liquid therethrough, but not of air, such that during a PCR the liquid sample remains in the reaction chamber (33) and after the PCR the reaction vessel (20) is configured to force the liquid sample via the spacer element (42) to the porous membrane (51) for hybridization and detection of the at least one target DNA in the liquid sample. Moreover, a PCR device comprising such a reaction vessel (20) as well as a method for performing PCR are described.
Core Innovation
The invention describes a reaction vessel (20) for performing a polymerase chain reaction (PCR) and detecting amplified PCR products, including a reaction chamber (33) and a storage chamber (63). The reaction chamber receives a liquid sample containing at least one target DNA, and a porous membrane (51) provides hybridization of the target DNA onto at least one specific hybridization probe immobilised on the porous membrane (51).
A spacer element (42) extends into the reaction chamber (33) from below the porous membrane (51) while being spaced from the bottom of the reaction chamber (33). The reaction chamber (33) and the storage chamber (63) are configured to be in fluid communication via the porous membrane (51) and a fluid channel defined by the spacer element (42), and the porous membrane is configured such that in a dry state it allows passage of air or other gases as well as liquid, while in a wet state it still allows passage of liquid but blocks passage of air or other gases.
The reaction vessel is configured such that during PCR the liquid sample remains in the reaction chamber (33), and after PCR the liquid sample can be forced via the spacer element (42) through the porous membrane (51) for hybridization and subsequent detection of the at least one target DNA in the liquid sample. The described device architecture enables forcing liquid through the porous membrane while the membrane dry/wet state provides gas-blocking behavior during liquid movement, and it supports subsequent optical detection of amplified target DNA.
Claims Coverage
The provided independent claims identify two main inventive claim sets for a PCR reaction vessel, each specifying the same core architecture: a reaction chamber and an overhead storage chamber, hybridization via a porous membrane with immobilised probes, and a spacer-defined internal fluid channel connecting the chamber volumes while controlling gas passage through a dry/wet porous membrane behavior.
Dry/wet porous membrane with immobilised hybridization probes
A porous membrane (51) for hybridization of the at least one target DNA within the liquid sample onto at least one specific hybridization probe immobilised on the porous membrane (51), wherein in a dry state the porous membrane allows passage of air or other gases as well as liquid therethrough and in a wet state still allows the passage of liquid therethrough but blocks the passage of air or other gases therethrough.
Reaction chamber and storage chamber connected by porous membrane and spacer-defined fluid channel
A reaction chamber (33) and a storage chamber (63) configured to be in fluid communication via the porous membrane (51) and a fluid channel defined by the spacer element (42).
Keep liquid in reaction chamber during PCR, then force through spacer and porous membrane for hybridization and detection
Configured such that during PCR the liquid sample remains in the reaction chamber (33) and after PCR the liquid sample can be forced via the spacer element (42) through the porous membrane (51) for hybridization and subsequent detection of the at least one target DNA in the liquid sample.
Internal fluid channel volume less than reaction chamber volume below spacer element
The internal fluid channel having a volume which is less than volume of the reaction chamber below the spacer element.
Across the independent claims, the inventive coverage is focused on (i) a porous membrane with dry/wet air-gas passage behavior combined with immobilised hybridization probes, (ii) fluid communication between a reaction chamber and an overhead storage chamber through the porous membrane and a spacer-defined internal fluid channel, and (iii) maintaining the liquid in the reaction chamber during PCR followed by forcing the liquid through the porous membrane after PCR for hybridization and subsequent detection.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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