System for releasing and isolating nucleic acids

Details System for releasing and isolating nucleic acids System for releasing and isolating nucleic acids

1998-02-17

2000-09-12

Ludlow, Jan

Chemical apparatus and process disinfecting, deodorizing, preser

Control element responsive to a sensed operating condition

209215, 209225, 210223, 422102, 422104, 435 6, 4352867, 436526, 536 254, B01L 1100

Patent

active

061173983

DESCRIPTION:

BRIEF SUMMARY
The object of the invention is a system for the release and isolation of nucleic acids and a procedure for using this system.
Detection procedures based on the determination of nucleic acids in a sample have increased in significance recently. This is due, among other things, to the high sensitivity of detection that these procedures can achieve. In terms of sensitivity, nucleic acid detection procedures are basically superior to antigen detection procedures. Although antigens are often relatively accessible in a sample, numerous steps are usually required to make nucleic acids accessible, especially when detecting organisms. In addition, nucleic acids are usually present in very low concentrations. Purification procedures for isolating nucleic acids from samples containing cells are known in particular, although they require a great deal of time and effort.
The sensitivity of the sample enrichment and pretreatment systems for nucleic acids currently on the market is often insufficient. In addition, automated sample pretreatment systems are not sufficiently safe from contamination to enable amplification, e.g. using the PCR. Another disadvantage of the automated sample pretreatment systems currently available is that they require the use of organic solvents (phenol and/or chloroform alcohol mixtures) to extract the nucleic acids.
The procedures in use today that immobilize nucleic acids basically use two principles to isolate nucleic acids. One principle calls for liquid samples containing nucleic acids to be aspirated through a solid phase which retains the nucleic acids. This step is preceded by a lysis step performed in a separate container. The nucleic acids are then dissolved from the solid matrix by aspirating an elution fluid through the matrix. The elation solution containing the nucleic acids is aspirated into a container for the next steps. It has been demonstrated, however, that the purity of the devices in use today does not meet the requirements for a subsequent amplification reaction, such as the PCR.
According to the second principle of nucleic acid isolation, the nucleic acids are removed by way of precipitation and then separated in a centrifuge. This procedure cannot be performed in a "batch" mode, however. Rather, it first requires that a solution containing cells be treated with lysing agents in a reaction vessel. The reaction mixture is then transferred by pipette from the container to a centrifugation tube. This tube contains an insert to which the released nucleic acids can adsorb, while the remaining fluid can flow to the bottom of the tube during centrifugation. The insert is treated one or more times with a fluid to wash the absorbed nucleic acids. For this step, the insert is transferred to a second centrifugation tube so that residues from the sample fluid do not reenter the insert. In the final step, the insert is placed in yet another container. An elution solution is then centrifuged through the insert to transfer the nucleic acids to another vessel that contains a solution that is capable of being processed further. This procedure is very susceptible to contamination, however, and requires transferring solutions between numerous reaction vessels.
The task of this invention was to provide a system for which the disadvantages of the state of the art are eliminated either completely or at least partially. In particular, this system can be used to absorb and desorb nucleic acids to a solid phase matrix without requiring a centrifuge for these steps.
A main feature of the invention is a receptacle for a sample processing vessel that can be kept at a constant temperature and set into motion in order to thoroughly mix the substances contained in the sample processing vessel. In addition, this receptacle is connected to a vacuum-generating system (e.g. a hose pump or a piston pump). The receptacle also makes it possible to separate magnetic particles within the sample processing vessel. Important advantages of the invention are the protection it provides against contamination (betwe

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patent: 5705062 (1998-01-01), Knubel

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