A kind of nucleic acid integration Multiple detection box body
Technical field
The utility model is related to a kind of nucleic acid detection chip of biology field, more particularly to a kind of nucleic acid integration
A variety of detection box bodys and detection method, belong to biomedical engineering and molecular diagnosis field.
Background technology
The research of current biological engineering in medicine is just deep into molecular level from entirety and cellular level.Nucleic acid is intracellular one kind
Important biomolecule participates in most of function of regulating cell, understands content of the nucleic acid in different cells with distribution to deeply
The biological significance for studying its function and behind is most important, to malignant tumour, AIDS, genetic disease and various infections
The checkout and diagnosis and treatment of disease are respectively provided with significance.Existing detection of nucleic acids mainly uses round pcr, and the method for PCR is sensitive
Degree is high, specificity is good, is current most common gene diagnosis method.But PCR method operate it is more complicated, and when testing
Between long, it is higher to personnel and instrument requirements, be not suitable for base or scene carry out quick diagnosis.
Isothermal amplification technique is a kind of current amplification technique relatively favored, and reaction process maintains constant always
At a temperature of, rapid amplifying is achieved the purpose that by the enzyme and respective specific primer that add different activities.With easy to operate,
The advantages that reaction time is short, high sensitivity is suitble to quick diagnosis field.But the efficient amplification efficiency of simultaneous isothermal amplification easily draws
Rise detected downstream when amplified production pollution, if when detecting by the sample process of upstream i.e. nucleic acid extraction, amplification and
The detection in downstream is enclosed in integration progress in a box body, then can effectively avoid the pollution of detection.While in order to adapt to base
Or the demand of field quick detection, the device detected is needed to minimize, it is portable.
To solve the above-mentioned problems, the utility model proposes a kind of nucleic acid integration Multiple detection box bodys.
Invention content
The utility model aim is for existing base or field quick detection demand, and it is multiple to provide a kind of nucleic acid integration
Box body is detected, which can realize the quick integral Multiple detection of nucleic acid, and compact, be convenient for carrying, can effectively prevent
The only pollution in detection process and the cross contamination of sample room.
In order to realize the purpose of this utility model and advantage, the utility model is achieved through the following technical solutions:
A kind of nucleic acid integration Multiple detection box body, box body include a shell and with two layers of runner plate in shell.
Two layers of runner plate mounted on top, can be put into shell.
The shell is rectangular, and one side of shell is opened wide, and another side is provided with an aperture, other each faces are closed.Institute
It states two layers of runner plate to be pushed into shell by open side, forms complete box body.
Two layers of runner plate is divided into top plate and lower plywood, and a plurality of flow channel for liquids and chamber are respectively carved in two-ply.
Preferably, the top plate sets nine chambers, wherein eight are hemispherical groove chambers, another is on upper strata
The chamber that plate embeds;Eight hemispherical groove chambers, wherein there are four being connected with runner, while with the top plate
Embedded chamber connection;Four additional hemispherical groove chamber does not connect with runner, respectively relatively independent.The top plate
The embedded chamber other end connects a bit of air pressure balance runner, and runner exit is communicated with shell one side aperture.
Preferably, the lower plywood is set there are five chamber, wherein four are hemispherical groove chambers, another is under
The chamber that laminate embeds;Four hemispherical groove chambers are connected with runner, while embed chamber with the lower plywood
One end is connected by runner;The lower plywood embeds the chamber other end and is connected with sample introduction runner one end, and the sample introduction runner is another
The extraneous sampling device of end connection.
The hemispherical groove chamber location of the top plate and the hemispherical groove chamber location of the lower plywood are mutual
It is corresponding;The hemispherical groove chamber notch of the top plate is downward, and the hemispherical groove chamber notch of the lower plywood is upward,
The chamber slot caliber size of two-ply is identical, and one-to-one correspondence can form fully spherical shape chamber.
Opposite slide can be carried out between the top plate and the lower plywood.Lower plywood first connects runner with top plate
Four hemispherical groove chambers, form four spherical chambers, and cause the embedded chamber of laminate up and down.With upper
The slip of laminate, lower plywood can form four spherical cavities with four relatively independent hemispherical groove chambers of top plate
Room.
It is preinstalled in four relatively independent hemispherical groove chambers of the top plate and is examined for nucleic acid isothermal amplification
The reagent solid dry powder of survey can be the detection of the various disease detection reagent or same target gene different loci of same sample
Reagent achievees the purpose that Multiple detection with this.
The sample introduction runner that extraneous sampling system first passes through lower plywood is passed through mineral oil to the embedded chamber of lower plywood, then lead to
Over-pressed Force system drives the mineral oil in chamber into the runner of connection, and drains lower plywood and embed mineral oil in chamber,
It drains into the embedded chamber of top plate;Sample to be tested and extracting solution are passed through to the embedded chamber of lower plywood by sampling system again,
The external world carries out heating and temperature control to embedded chamber, and reaction temperature is controlled at 100 DEG C, reacts 10 minutes, realizes the height of nucleic acid
Warm one-step method extraction.Top plate is slided, makes four relatively independent hemispherical groove chambers of lower plywood and top plate, shape
Into four spherical chambers.By in the liquid driven after being reacted in process two to four spherical chambers, and full of chamber, dissolve pre- envelope
Mounted in the reagent solid dry powder of the indoor nucleic acid isothermal amplification detection of top plate hemispherical groove chamber, external world's auxiliary mixing reagent.
Sampling system is passed through mineral oil to the embedded chamber of lower plywood again, and drives into the runner of lower plywood.To the four of top plate
A relatively independent hemispherical groove chamber carries out heating and temperature control, and reaction temperature is controlled at 65 DEG C, reacts 30 minutes.Instead
Fluorescence detection device should be answered to carry out the real-time detection of reactant in the process.It is shown after reaction by observing fluorescence detection device
Each chamber fluorescence curve, judge the result of Multiple detection.
The beneficial effects of the utility model:
The utility model use box body layered structure, by entire detection of nucleic acids process integration integration closing ring
Border, by high temperature one-step method nucleic acid extraction, isothermal duplication, real-time fluorescence detection is combined, and highly shortened traditional detection of nucleic acids
Time, and the cost requirement of equipment is substantially reduced, effectively prevents extraneous pollution and the cross contamination of nucleic acid;This reality
With novel, box body difficulty of processing is smaller, easy to operate portable, meets the needs of Site Detection.
Description of the drawings
Fig. 1 the utility model construction profile schematic diagrames.
Fig. 2 the utility model structure perspective diagrams.
Fig. 3 the utility model upper strata plate structure schematic diagram.
Fig. 4 the utility model lower floor plate structure schematic diagram.
Laminate initial phase is to position vertical view (A) and side view (B) above and below Fig. 5 the utility model.
Lower plywood nucleic acid amplification detection process relative position vertical view (A) and side view (B) in Fig. 6 the utility model.
Wherein:1- box bodys, 2- shells, 3- top plates, 4- lower plywoods, 301- upper stratas runner, the chamber that 302- top plates embed
Room, 303- upper stratas runner communication groove chamber, 304- separate chambers, 305- balanced runners, 401- lower floors runner, 402- lower plywoods
Embedded chamber, 403- lower floors runner communication groove chamber, 404- sample introduction runners.
Specific embodiment
The utility model is done with reference to embodiment and attached drawing 1-6 and is further explained, to enable people in the art
Member with reference to specification word can basis to implement.The following example is merely to illustrate the utility model, but is not used to limit this
The practical range of utility model.
A kind of nucleic acid integration Multiple detection box body and detection method, as illustrated in fig. 1 and 2, box body 1 include a shell 2,
And two layers of runner plate (top plate 3 and lower plywood 4) in shell 2.Two layers of runner plate mounted on top, can be put into shell.Institute
It is rectangular to state shell 2, one side is opened wide, and another side is provided with an aperture, other each faces are closed.Two layers of runner plate
It can be pushed into shell by open side, form complete box body 1.A plurality of liquid is respectively carved in the top plate 3 and lower plywood 4
Body runner and multiple chambers.
It is the structure diagram of the top plate 3 as shown in Figure 3, there is nine chambers and several upper strata runners 301.Wherein one
A is the chamber 302 embedded in top plate, and in addition eight are hemispherical groove chambers.Wherein the four of eight hemispherical groove chambers
A connection runner and the chamber 302 that embeds of top plate, referred to as upper strata runner communication groove chamber 303, four additional not with stream
Road connects, respectively relatively independent, referred to as separate chamber 304.302 other end of chamber that the top plate embeds connects one section of air pressure
Balanced runner 305.
It is the structure diagram of the lower plywood 4 as shown in Figure 4, there are five chamber and several lower floor's runners 401.Wherein one
A is the chamber 402 embedded in lower plywood, and four additional is hemispherical groove chamber, connects the chamber that runner and lower plywood embed
Room 402, referred to as lower floor's runner communication groove chamber 403.402 other end of chamber that the lower plywood embeds connects one section of sample introduction stream
Road 404.
It can carry out sliding relatively between the top plate 3 and lower plywood 4, the runner communication groove chamber of top plate 3
303 and separate chamber 304 notch it is downward, runner communication groove 403 notches of chamber of lower plywood 4 are upward, the chamber of two-ply
Notch caliber size is identical, and one-to-one correspondence can form fully spherical shape chamber.
For above-mentioned nucleic acid integration Multiple detection box body, method used below is used in case:
The top plate 3 and lower plywood 4 is stacked together before being put into shell 2, in the separate chamber 304 of top plate 3
The reagent solid dry powder that prepackage detects for nucleic acid isothermal amplification can be the various disease detection reagent or same of same sample
The detection reagent of one target gene different loci, achievees the purpose that Multiple detection with this.
Before the experiment of nucleic acid integration Multiple detection, the initial phase contraposition of the top plate 3 and lower plywood 4 in shell 2
It puts as shown in figure 5, upper strata runner communication groove chamber 303 and 403 notch of lower floor's runner communication groove chamber correspondence, form four
Spherical chamber, and connect each channel.When experiment starts, the sample introduction runner 404 that extraneous sampling system first passes through lower plywood 4 is passed through
Mineral oil then by pressure system is driven the mineral oil in embedded chamber 402 to connection to the embedded chamber 402 of lower plywood 4
Runner 401 in, and empty lower plywood and embed mineral oil in chamber, drain into the embedded chamber 302 of top plate 3 and upper strata runner
In 301.
Sample to be tested and extracting solution are passed through to the embedded chamber 402 of lower plywood 4 by sampling system again, it is extraneous to embedding chamber
Room 402 carries out heating and temperature control, and reaction temperature is controlled at 100 DEG C, reacts 10 minutes, realizes that the high temperature one-step method of nucleic acid carries
It takes.
After the completion of extraction, top plate 3 is slided, makes the four of 4 runner communication groove chamber of lower plywood, 403 notch and top plate 3
A 304 notch of separate chamber corresponds to connection, forms four spherical chambers.By the nucleic acid liquid extracted embedding from lower plywood 4
Chamber 402 is driven into four spherical chambers, and full of chamber, is dissolved pre-packaged in the indoor core of top plate hemispherical groove chamber
The reagent solid dry powder of sour isothermal duplication detection, external world's auxiliary mixing reagent.
Sampling system is passed through mineral oil toward the embedded chamber 402 of lower plywood 4 again, and drives to the runner 401 of lower plywood
In, spherical chamber is closed, prevents from evaporating under the reagent condition of high temperature.Heating temperature is carried out to four separate chambers 304 of top plate 3
Control controls reaction temperature at 65 DEG C, reacts 30 minutes.Fluorescence detection device is answered to carry out the real-time of reactant in reaction process
Detection.The fluorescence curve of each chamber shown after reaction by observing fluorescence detection device judges the knot of Multiple detection
Fruit.
High temperature one-step method nucleic acid extraction, isothermal duplication, real-time fluorescence detection are combined, greatly shortened by the utility model
The time of traditional detection of nucleic acids, and the cost requirement of equipment is substantially reduced, entire detection of nucleic acids process integration one
Change is completed in the environment of closing, effectively prevents extraneous pollution and the cross contamination of nucleic acid, easy to operate portable, very full
The demand of sufficient Site Detection.