During concrete enforcement, the determination of the pumping signal parameter in testing laboratory in completing steps 1, at the reaction member that factory's completing steps 2 obtains, and completes the corresponding antigen of target antibody or antigen or the bag quilt of antibody.In step 4, pre-service is carried out to humoral sample to be measured, contaminating cell can be prevented the impact of test result, improve the accuracy of test result.In step 5, pumping signal is applied to reaction member, dielectrophoresis effect and electrocaloric effect is produced in reaction member, dielectrophoresis effect makes the target antibody in humoral sample to be measured or antigen move to electrode slice direction, corresponding antigen or the antibody of accelerating target antibody or antigen and electrode slice pan coating combine, thus making the thickness of the electrostatic double layer of junction electrode surface, thickening (originally the electrostatic double layer thickness of electrode surface is the antigen of bag quilt or the thickness of antibody, and present electrostatic double layer thickness is target antibody or antigen with corresponding antigen or antibody to the thickness after combining.)。Along with target antibody or antigen and corresponding antigen or antibody more to the quantity combined, the area of junction increases, and the thickening electrostatic double layer area of thickness constantly increases, and makes the total resistance change of reaction member inside greatly, specifically, in impedance, capacitive reactance composition will be more and more less.Can qualitatively draw testing result and upload high in the clouds.
Like this, in testing process, apply pumping signal, produce electrophoretic effect and accelerate target antibody or the antigen speed to the movement of electrode slice direction; Produce electrocaloric effect make reaction member inside formed significant thermograde and the local fluid flow that causes by with dielectrophoresis effect together accelerate the corresponding antigen of target antibody or antigen and electrode slice pan coating or antibody in conjunction with speed.Like this, with traditional reacting phase ratio, often only need can complete reaction in 1 ~ 5 minute, improve the efficiency of detection.Secondly, utilize double electrode layer theory, measure the change of the resistance value of reaction member inside, then resistance value is analyzed, testing result can be drawn qualitatively, require relatively low to the specialty of testing staff, use convenient.Again, testing result is uploaded high in the clouds, be conducive to the integration of data, can persistence, be convenient to systematic record.
As optimization, the check-out console of the reaction member that step 2 obtains also is coated with one deck sealer; Sealer generally adopts unrelated protein to prepare, and object is that the position that adopts unrelated protein to occupy in reaction member not occupied by antigen or antibody is to reduce non-specific adsorption.Prevent from other species adsorbs to electrode slice impact resistance value, like this, the accuracy of detection can be improved.
As optimization, the method of sampling of step 5) middle impedance value is: after applying pumping signal, every 10 ~ 900 milliseconds of record resistance values of primary first-order equation unit inside of impedance signal receiver module test constantly 1 ~ 5 minute, be then stored into the resistance value of record in memory module.Like this, in the process of whole reaction, every 10 ~ 900 milliseconds of records resistance value, can obtain more sampled data, is conducive to detecting the accuracy improving testing result.
As optimization, the method of sampling of step 5) middle impedance value is: before the corresponding antigen of target antibody or antigen and bag quilt or antibody combine, the signal of 10Hz-10MHz is adopted to perform sweep test, and record 10-100 group resistance value, then apply pumping signal and continue 1 ~ 5 minute, the last signal of 10Hz-10MHz that again adopts performs sweep test, and records 10-100 group resistance value.
Like this, adopt the signal of 10Hz-10MHz to perform the biosystem impact of sweep test on reaction member inside humoral sample to be measured during sampling little, make the resistance value of sampling more accurate.Apply the pumping signal of target antibody or antigen during reaction, the antibody antigen engaging speed of reaction member inside can be accelerated, like this, while ensure that detection speed, improve accuracy in detection.
As optimization, step 2) electrode slice in the reaction member that obtains comprises the electrode body in strip that two panels is parallel to each other, electrode body is positioned at the electrode branch near the side of another electrode body with the strip be parallel to each other of some equidistant settings, and the other end of electrode branch is close to another electrode body; The interlaced setting of electrode branch of two electrode bodies.
Like this, in unit area, the length of electrode slice relative edge can be increased, make electrophoresis and electrocaloric effect more strong, fast reaction speed, be conducive to improving detection efficiency.Meanwhile, the equivalent electrical circuit of electrode shape and correspondence is relatively simple, handling ease, low cost of manufacture.
As further optimization, the communication module in the auxiliary control module that step 3) obtains comprises a USB interface and a bluetooth module.
Like this, intelligent movable equipment can carry out communication by data line and communication module, also by bluetooth and communication module communication, can have fault tolerance.Meanwhile, USB interface can also be used for, for processor module firmware updating is to improve detected parameters, making it can mate more eurypalynous reaction member.
Based on an antibody antigen detection method for mobile device, it is characterized in that, comprise the steps:
1) the pumping signal parameter of target antibody or antigen is determined; The signal of 10Hz ~ 10MHz is adopted to carry out full scan to target antibody or antigen in the lab, the response condition of object observing antibody or antigen and find target antibody or antigenic response the strongest time signal parameter, be defined as the pumping signal parameter of target antibody or antigen;
2) obtain the reaction member of following structure, comprise an open-topped reaction chamber, be provided with one piece of check-out console, check-out console be equipped with at least one pair of electrode slice bottom reaction chamber, the terminals of electrode slice pass and are fixed on the box body of reaction chamber; Check-out console is also fixed with corresponding antigen or the antibody of target antibody or antigen;
3) obtain the auxiliary control module of following structure, comprise processor module, the communication module be electrically connected with processor module, memory module, impedance signal receiver module and signal generator; The output terminal of impedance signal receiver module is connected to the impedance signal input end of processor module; The parameters input end of signal generator is electrically connected to the parameter output of processor module;
Also comprise for Connection Step 2) measurement interface of the terminals of reaction member that obtains, one end of measurement interface connects the input end of described impedance signal receiver module, and the other end of measurement interface is connected with a pumping signal conditioning module; Pumping signal conditioning module has one for carrying out the data-interface of Signal transmissions with intelligent movable equipment earpiece holes;
Pumping signal conditioning module comprises the signal conditioning circuit being arranged on input end, signal frequency multiplication circuit and be arranged on the signal amplification circuit of output terminal;
Impedance signal receiver module comprises the current-voltage modular converter being arranged on input end, low-pass filter and be arranged on the analog to digital converter of output terminal;
4) sampling of humoral sample to be measured and pre-service, obtains humoral sample to be measured and by the cell filtering in humoral sample to be measured;
5) sampling of pumping signal and resistance value is applied, the link of reaction member step 2 obtained is connected on the measurement interface of the auxiliary control module that step 3 obtains, again by pretreated humoral sample instillation reaction member to be measured, and contact with the electrode slice on the check-out console in reaction member; The earpiece holes of intelligent movable equipment is wired on the data-interface of auxiliary control module, the pumping signal parameter of the target antibody determined according to step 1 of the TT&C software of intelligent movable equipment or antigen again, and export sinusoidal signal by earpiece holes, to pumping signal conditioning module, pumping signal is applied to electrode slice.
Meanwhile, impedance signal receiver module detects and records the change in impedance value of reaction member inside, is stored in memory module by the resistance value of record;
6) data analysis draw testing result; Adopt intelligent movable equipment connection to the communication module of auxiliary control module, and read the resistance value recorded in auxiliary control module, being analyzed by the resistance value of record by being arranged on the TT&C software in intelligent movable equipment, testing result can be drawn qualitatively; Testing result is uploaded high in the clouds by network and is filed by last intelligent movable equipment.
Like this, the TT&C software of intelligent movable equipment can be adopted according to different target antibodies or antigen setting pumping signal parameter, and to pumping signal conditioning module, pumping signal is applied to electrode slice by earpiece holes output sinusoidal signal.Compared to processor module setup parameter, intelligent movable equipment can make the output of the setting of parameter and pumping signal convenient, more flexibly.TT&C software can be downloaded corresponding pumping signal parameter according to different target antibodies or antigen from high in the clouds and be used for setting, and processor module can only prior burned limited pumping signal parameter, as needed change, also need to use computer and cd-rom recorder to operate by professional.Like this, this system can adapt to different target antibodies or antigen more fast.
In sum, the antibody detection method based on mobile device of the present invention has and is easy to operation, and detection speed is fast, and accuracy of detection is advantages of higher comparatively.
Embodiment
The present invention is described in further detail to detect test macro and accompanying drawing below in conjunction with a kind of antibody antigen adopting the inventive method.
Embodiment 1
As depicted in figs. 1 and 2, a kind of antibody antigen detects test macro, comprise at least one reaction member 1, an open-topped reaction chamber drawn together by reaction member bag 1, one piece of check-out console is provided with bottom reaction chamber, check-out console is equipped with at least one pair of electrode slice 11, the terminals 12 of electrode slice 11 pass and are fixed on the box body of reaction chamber; Check-out console is also fixed with corresponding antigen or the antibody of target antibody or antigen.
The corresponding antigen of fixed target antibody or antigen or antibody are namely to antigen
or the bag of antibody is processed.Can better be fixed on check-out console after the corresponding antigen of the target antibody in humoral sample to be measured or antigen and electrode slice pan coating or antibody can be made to combine, be conducive to improving the accuracy detected.
After humoral sample instillation reaction chamber to be measured, electrode slice forms electrostatic double layer, after antibody antigen engages, change the thickness of electrostatic double layer, the resistance value of reaction member inside is changed, by the change size of measurement impedance value, just can draw testing result.
Wherein, also comprise an auxiliary control module 2, auxiliary control module 2 comprises power module (not shown) and processor module 21, the communication module 22 be electrically connected with processor module 21, memory module 23, impedance signal receiver module 24 and signal generator 25; The output terminal of impedance signal receiver module 24 is connected to the impedance signal input end of processor module 21; The parameters input end of signal generator 25 is electrically connected to the parameter output of processor module 21; Auxiliary control module 2 also comprises the measurement interface of the terminals 12 for connecting described reaction member 1, one end of measurement interface connects the input end of described impedance signal receiver module 24, the other end of measurement interface is connected with a pumping signal conditioning module 26, the output terminal of the input end connection signal generator 25 of pumping signal conditioning module 26; Pumping signal conditioning module 26 comprises the signal conditioning circuit 261 being arranged on input end, signal frequency multiplication circuit 262 and be arranged on the signal amplification circuit 263 of output terminal; Impedance signal receiver module 24 comprises the current-voltage modular converter 241 being arranged on input end, low-pass filter 242 and be arranged on the analog to digital converter 243 of output terminal; Also comprise the intelligent movable equipment 3 that one is provided with TT&C software, described intelligent movable equipment 3 is connected on the communication module 22 of described auxiliary control module 2.
During detection, first, the measurement interface of the terminals of reaction member with auxiliary control module is connected, by humoral sample to be measured from the open top instillation reaction chamber of reaction member, and contacts with the electrode slice on check-out console.Restart auxiliary control module, processor module is according to target antibody or antigen setting pumping signal parameter, occurred to produce by signal generator and passed through pumping signal conditioning module and pumping signal is applied to the electrode slice of reaction member, like this, dielectrophoresis effect and electrocaloric effect is produced in reaction member, dielectrophoresis effect makes the target antibody in humoral sample to be measured or antigen move to electrode slice direction, corresponding antigen or the antibody of accelerating target antibody or antigen and electrode slice pan coating combine, thus making the thickness of the electrostatic double layer of junction electrode surface, thickening (originally the electrostatic double layer thickness of electrode surface is the antigen of bag quilt or the thickness of antibody, present electrostatic double layer thickness is target antibody or antigen with corresponding antigen or antibody to the thickness after combining.)。Along with target antibody or antigen and corresponding antigen or antibody more to the quantity combined, the area of junction increases, and the thickening electrostatic double layer area of thickness constantly increases, and makes the total resistance change of reaction member inside greatly, specifically, in impedance, capacitive reactance composition will be more and more less.
Impedance signal receiver module detects and records the change in impedance value of reaction member inside, is stored in storage unit by the resistance value of record.Then the resistance value recorded in storage unit is sent in intelligent movable equipment by communication module, and intelligent movable equipment is analyzed the resistance value received by TT&C software, can draw testing result qualitatively.Testing result is uploaded high in the clouds by network by last intelligent movable equipment, does process of filing.
Apply pumping signal in testing process, produce electrophoretic effect and accelerate target antibody or the antigen speed to the movement of electrode slice direction; Produce electrocaloric effect make reaction member inside formed significant thermograde and the local fluid flow that causes by with dielectrophoresis effect together accelerate the corresponding antigen of target antibody or antigen and electrode slice pan coating or antibody in conjunction with speed.Like this, with traditional reacting phase ratio, often only need can complete reaction in 1 ~ 5 minute, improve the efficiency of detection.Secondly, utilize double electrode layer theory, measure the change of the resistance value of reaction member inside, then resistance value is analyzed, testing result can be drawn qualitatively, require relatively low to the specialty of testing staff, use convenient.Again, testing result is uploaded high in the clouds, be conducive to the integration of data, can persistence, be convenient to systematic record.The equipment volume that whole detection system adopts is little, is easy to carry, and facilitates testing staff's spot sampling to detect, can realize Real-time Obtaining testing result, improve detection speed.
During concrete enforcement, auxiliary control module can also arrange many group pumping signal conditioning module and impedance signal receiver module, like this, can detect simultaneously, and detect different antibody or antigen for different reaction members to multiple reaction member.
Wherein, described electrode slice 11 comprises the electrode body in strip that two panels is parallel to each other, electrode body is positioned at the electrode branch near the side of another electrode body with the strip be parallel to each other of some equidistant settings, and the other end of electrode branch is close to another electrode body; The interlaced setting of electrode branch of two electrode bodies.
Like this, in unit area, the length of electrode slice relative edge can be increased, make electrophoresis and electrocaloric effect more strong, fast reaction speed, be conducive to improving detection efficiency.Meanwhile, the equivalent electrical circuit of electrode shape and correspondence is relatively simple, handling ease, low cost of manufacture.During concrete enforcement, electrode slice can also adopt asymmetric electrode pair, also can adopt mesh electrode, can also adopt other electrode form various such as spiral, castle electrode.
Wherein, described check-out console is also equipped with one deck sealer.Sealer generally adopts unrelated protein to prepare, and object is that the position that adopts unrelated protein to occupy in reaction member not occupied by antigen or antibody is to reduce non-specific adsorption.Prevent from other species adsorbs to electrode slice impact resistance value, like this, the accuracy of detection can be improved.
Wherein, described pumping signal conditioning module 26 also has one for carrying out the data-interface of Signal transmissions with described intelligent movable equipment 3 earpiece holes; Described data-interface is wired in the earpiece holes of described intelligent movable equipment 3.
Like this, the TT&C software of intelligent movable equipment can be adopted according to different target antibodies or antigen setting pumping signal parameter, and to pumping signal conditioning module, pumping signal is applied to electrode slice by earpiece holes output sinusoidal signal.Compared to processor module setup parameter, intelligent movable equipment can make the output of the setting of parameter and pumping signal convenient, more flexibly.TT&C software can be downloaded corresponding pumping signal parameter according to different target antibodies or antigen from high in the clouds and be used for setting, and processor module can only prior burned limited pumping signal parameter, as needed change, also need to use computer and cd-rom recorder to operate by professional.Like this, this system can adapt to different target antibodies or antigen more fast.
Wherein, the communication module 22 in described auxiliary control module 2 comprises a USB interface and a bluetooth module.Like this, intelligent movable equipment can carry out communication by data line and communication module, also by bluetooth and communication module communication, can have fault tolerance.Meanwhile, USB interface can also be used for, for processor module firmware updating is to improve parameter, making it can mate more eurypalynous reaction member.
During concrete enforcement, communication module can also adopt other module making such as NFC, Lightning.
Embodiment 2
As shown in Figure 3-Figure 5, a kind of detection method adopting the antibody antigen of embodiment 1 to detect test system and test mastitis, adopts following steps:
1) the pumping signal parameter of target antibody or antigen is determined; The signal of 10Hz ~ 10MHz is adopted to carry out full scan to target antibody or antigen in the lab, the response condition of object observing antibody or antigen and find target antibody or antigenic response the strongest time signal parameter, be defined as the pumping signal parameter of target antibody or antigen;
During concrete enforcement, as shown in Figure 3, the signal of 10Hz ~ 10MHz is adopted to scan the experimental liquid containing mastitis antibody, rate of change of capacitance in detection experiment liquid, find when signal frequency is when more close to 100kHz, the rate of change of capacitance in experimental liquid is larger, therefore, determine that the amplitude of the pumping signal of mastitis antibody is 1V, frequency is 100kHz.
2) acquisition is prior is coated with mastitis pathogenic bacterium antigen in factory and does the reaction member of Seal treatment, and is connected with the measurement interface of terminals with auxiliary control module by the link of reaction member.
3) adopt blood taking needle to obtain blood sample to be measured, and by blood filter by the cell filtration in blood, prevent cell from having an impact to testing result.Then by the reaction chamber of blood sample instillation reaction member to be measured, and with the check-out console bottom reaction chamber to contacting.
4) earpiece holes being provided with the mobile phone of TT&C software be connected with the data-interface of auxiliary control module, and the amplitude arranging pumping signal in mobile phone TT&C software is 1V, frequency is 100kHz.Open the power supply of auxiliary control module, mobile phone TT&C software sends pumping signal by earpiece holes, and is applied to reaction member by pumping signal conditioning module.
During concrete enforcement, the pumping signal parameter of all right prior burned mastitis antibody in processor module, then produces signal by processor die block control signal generator and is applied to reaction member by pumping signal conditioning module.
5) impedance signal receiver module detects and recorded the capacitance variation of reaction member inside every 1 second, is stored in storage unit by the capacitance of record.Then the capacitance recorded in storage unit is sent in mobile phone by communication module, and mobile phone is analyzed by the capacitance of TT&C software by reception, can draw testing result qualitatively.
As shown in Figure 4, Fig. 4 gives the positive normalization capacitance variations situation detected with negative breast inflammation, and tests sample in contrast with damping fluid (Buffer-B).Blood sample in figure dilutes according to the dilution ratio of 1:1, and as can be seen from the figure, blood to be measured is positive, and contrast test sample is not because containing mastitis antibody, and be negative reaction.
As shown in Figure 5, in Fig. 5, blood sample is divided into 5 parts, this dilutes according to the dilution ratio of 1:1,1:20,1:40,1:80 and 1:120 respectively, the capacitance variation curve obtained after 1s samples, as can be seen from the figure, the content of mastitis antibody is higher, it is faster that capacitance declines, when blood sample dilutes according to the dilution ratio of 1:120, within the detection time of 3 minutes, be difficult to pick out the difference with the capacitance of negative control group.This shows, qualitative analysis can go out antibody content in blood according to the change of capacitance.
During concrete enforcement, can also realize carrying out data scaling to target antibody, then adopt detected value and calibration value to contrast, like this, quantitatively can draw the target antibody content in testing sample.