JP2003000226A - Cell disintegration device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To solve the problem of conventional continuous PCR process comprising the difficulty in getting sufficient disintegration effect on cell membrane of leukocyte, bacterium, virus, or the like, for DNA analysis and the actions of a reagent and an enzyme as PCR inhibiting substances. SOLUTION: Fine magnetic beads and metal balls, or the like, are used as a means for breaking cell membrane. Cells can be broken by vibrating, moving or heating the magnetic beads and metal balls by applying a dynamically varying magnetic field to the magnetic beads and metal balls mixed in the specimen to be examined.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an invention in the field of biotechnology, and relates to crushing of living cells required as a pretreatment step such as a PCR method in DNA analysis.

[0002]

2. Description of the Related Art In order to extract DNA from cells, it is first necessary to disrupt target cells, that is, cell membranes of white blood cells, bacteria, viruses and the like. A surfactant for this purpose,
Physical stimuli such as crushing with reagents such as acids and alkalis, proteolytic enzymes, heat, ultrasonic waves, or centrifugal separation of samples mixed with glass beads have been used.

However, depending on the conditions such as the type of sample to be disrupted, in many cases cell membrane disruption efficiency cannot be sufficiently obtained. When reagents or enzymes are used, the reagents or enzymes act as PCR inhibitors, In this state, the PCR method (polymerase cha) is a continuous operation.
There has been a problem that in-reaction cannot be performed. Therefore, there has been a demand for a method of efficiently disrupting the cell membrane without inhibiting the PCR process.

[0004]

DISCLOSURE OF THE INVENTION In the present invention, there is provided an inexpensive cell membrane disruption method capable of effectively disrupting cell membranes of leukocytes, bacteria, viruses and the like, which does not inhibit the PCR process in the next step. Is to provide.

[0005]

In the present invention, fine magnetic spheres, metal spheres, etc. are used as means for destroying cell membranes. By applying a dynamically changing magnetic field to a magnetic sphere, a metal sphere, or the like mixed with the sample to be inspected, the magnetic sphere or the metal sphere is vibrated, moved, or heated to destroy the cells. Other than thermophilic bacteria, metal spheres may not be used to avoid fever.

The present invention is a method or apparatus for crushing cells, characterized in that fine magnetic spheres or metal spheres are placed in and mixed with a test sample of the white blood cells, bacteria, viruses, etc. placed in a test container. , A portion for accommodating and fixing the test container, a plurality of electromagnet groups for applying magnetic fields to the test container from multiple directions, and a control unit for randomly driving the electromagnet group.

White blood cells, bacteria, in test containers
A DNA analysis sample such as a virus is put into an apparatus after being sufficiently mixed with fine magnetic spheres or metal spheres. The magnetic spheres and metal spheres are according to the cell size of the test sample,
The optimum size needs to be chosen.

The electromagnet group can be energized at a random timing, and at the same time, the direction of the energizing current can be switched to reverse the magnetic poles if necessary. by this,
The magnetic sphere moves or rotates irregularly.
Further, the metal spheres generate heat and may move or rotate depending on how the magnetic field is applied. The cells of the test sample are easily disrupted by the movement or rotation of the magnetic spheres or the heat, movement, or rotation of the metal spheres.

The method of recovering the magnetic spheres and the metal spheres after the crushing of the cells is completed can be easily performed by inserting a rod-shaped permanent magnet into the sample container.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of the present invention will be described based on examples with reference to the drawings. FIG. 1 is a conceptual diagram of the present invention. 1 and 2 are electromagnets, 3 is a sample container, 4 is DNA.
A sample container, a magnetic ball, and a metal ball are stored in a sample container. When the electromagnet is energized, a magnetic field is generated, a force is applied to the magnetic sphere or the metal sphere, and movement, rotation, heat generation, etc. occur.

FIG. 2 shows an embodiment of the present invention. In the present invention, the number of electromagnets is arbitrary, but the figure shows an example in which four electromagnets are used. In the figure, 5 is a DNA sample container, 6, 7, 8 and 9 are electromagnets, and 10 and 1
Reference numerals 1, 12, and 13 are transmitters that are connected to the electromagnets 6 to 9 and drive the electromagnets. Reference numeral 14 is a control unit, which functions to control the output timing of the oscillators 10 to 13.

FIG. 3 is a waveform diagram of the oscillators 10 to 13 in FIG. As is clear from the figure, the electromagnet 6
The driving electric signals applied to the signals Nos. 9 to 9 can take arbitrary timings, and the timings are selected so as to most effectively drive the magnetic spheres or metal spheres mixed in the sample container.

Similarly to FIG. 3, FIG.
FIG. 13 is a waveform diagram of the oscillators from Nos. In FIG. 3, the direction of the current flowing through each electromagnet is fixed, but the control waveform in FIG. 4 has a mechanism that not only turns the current on and off but also reverses the polarity, enabling more effective control. ing.

[0014]

EFFECTS OF THE INVENTION Regarding the disruption of living cells required as a pretreatment for DNA analysis, the efficiency of disruption of the cell membrane has not been sufficient so far, or even if the disruption effect exists, the next step of PC
Although there was a problem such as inhibiting the R process, by using the present invention, it is possible to effectively disrupt cell membranes of leukocytes, bacteria, viruses and the like, and not disrupt the PCR process of the next step, cell membrane disruption. Has become possible.

[Brief description of drawings]

[Figure 1] Conceptual diagram of cell disruption device

[Fig. 2] Example of cell disruption device

FIG. 3 Example of oscillator waveform driving an electromagnet

FIG. 4 is another example of the oscillator waveform for driving the electromagnet.

Claims (2)

[Claims] 1. A DNA analysis sample of leukocytes, bacteria, viruses, etc. is mixed with fine magnetic spheres and, if necessary, fine metal spheres, etc., and the DNA analysis sample is regularly or irregularly applied from multiple directions. By applying a magnetic field, motion such as vibration, rotation, movement, etc. to the magnetic spheres and metal spheres, as well as giving heat by electromagnetic induction as needed, crush the cells of the white blood cells, bacteria, viruses etc. Cell crushing method. 2. In order to realize claim 1, a portion for accommodating a container containing the DNA analysis sample is provided, and the D
It has a plurality of electromagnet groups arranged to apply a magnetic field to a container containing the NA analysis sample, and the electromagnet group has arbitrary timing and arbitrary polarity in order to effectively disrupt the cells of the DNA analysis sample. 1. A cell disruption device for DNA analysis, comprising a control unit that can be driven by.