CN110564657A - Microbial sensor for early detection of breast cancer and detection system thereof - Google Patents
Microbial sensor for early detection of breast cancer and detection system thereof Download PDFInfo
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Abstract
The invention provides a microbial sensor for early detection of breast cancer and a detection system thereof, wherein a multi-target miRNA biomarker joint detection technology based on a genetic engineering microbial sensor is characterized in that a genetic circuit of a logic AND gate is formed by utilizing the action site characteristics of integrase fimE and integrase BxB1 through a means of genetic recombination, genetic engineering escherichia coli capable of realizing the joint detection of miRNA-21 and miRNA-155 is constructed, and the simultaneous detection of two biomarkers is realized by detecting a fluorescent signal of green fluorescent protein expressed by the escherichia coli in cooperation with a microfluidic chip and a portable fluorescent detection system.
Description
Technical Field
The invention belongs to the field of biomedical detection, and particularly relates to a microbial sensor for early detection of breast cancer and a detection system thereof.
Background
The breast cancer is one of the most common malignant tumors of women, the morbidity and the mortality rate are the first of various malignant tumors of women, and the health of the women is seriously threatened. In recent years, the treatment of breast cancer has been improved, but it is a prerequisite that the breast cancer can be found and treated in time.
The following methods are commonly used for detecting breast cancer: the touch method can screen a part of patients by touching with an experienced doctor. Meanwhile, patients can also perform self-examination through breast palpation, however, the latest breast cancer screening guidelines issued by the american society of gynecologists and obstetricians believe that the breast palpation self-examination of patients cannot reduce the detection rate and mortality rate of breast cancer, and instead, due to over-examination, normal breast and isolated nodules are easily mistaken for breast cancer (false positive), resulting in panic and even excessive surgical treatment. Ultrasonic examination method: the ultrasonic examination can display the size, the shape, the boundary, the blood flow signal and the like of the tumor, and is suitable for women of all ages. However, it may have certain limitations for small lesions or micro calcifications. Soft X-ray molybdenum target photography: the molybdenum target has higher inspection accuracy and can display micro calcified lesions. The molybdenum target examination is more suitable for married and fertile women over 30 years old. However, the examination of the dense mammary gland, the mammary gland with the prosthesis implanted and the mammary gland with the epilepsy formed after the treatment is difficult to carry out. Nuclear magnetic resonance method: although the nuclear magnetic resonance has higher examination accuracy, the benign and malignant breast tumors can be objectively evaluated, but the examination cost is high, the requirement on indications is higher, the radiation degree is higher, and the nuclear magnetic resonance cannot be generally used as a routine physical examination item.
The detection methods are difficult to diagnose the breast cancer at an early stage, so that the early diagnosis of the breast cancer through the biomarkers has very important value and significance in the future stages with unobvious pathological characteristics.
The breast cancer biomarkers comprise estrogen receptors, kallikrein 10 and the like, meanwhile, gene expression abnormality caused by epigenetic change is also an important factor for cancer occurrence, and the gene expression can be correspondingly changed in the cancer occurrence and development stages. miRNA is endogenous single-stranded small molecular RNA with the length of 21-25 bases, and the specificity expressed in tumor cells plays an important role in the processes of generation, development, infiltration, metastasis, prognosis and the like of tumors. Research shows that the content of miRNA-21 and miRNA-155 in the breast cancer future stage is changed, so that the combined detection of the two miRNAs is realized, and the method plays a very key role in early screening of the breast cancer.
Most of the existing detection methods for protein breast cancer biomarkers are based on immune reactions between antigen and antibody, but due to instability of the antibody, certain deviation sometimes occurs in detection results. Therefore, the detection of miRNA small molecules as biomarkers of breast cancer becomes a hot spot of current research. To date, many studies have reported analysis methods for miRNA detection, among which Northern blotting, microarray analysis, and real-time quantitative polymerase chain reaction (qRT-PCR) are considered as conventional standard methods, but still have some disadvantages such as large sample demand, erroneous amplification resulting in preference, and susceptibility to false positive results.
The microbial sensor has the characteristics of small volume and short response time, and the sensor identification molecule formed by microorganisms with better specificity does not need pretreatment generally for a sample to be detected, and meanwhile, the growth process of the microorganisms plays a certain signal amplification role, so that the application of the microbial sensor to biological detection becomes one of the hot spots of the current research. On the basis of a microbial sensor, the sensor can be further modified by combining a genetic engineering means, so that the sensor can respond to a target substance in the environment, further stimulate the expression of a series of downstream genes and generate a specific signal which can be captured, thereby completing detection.
The detection of a single biomarker has difficulty in accurately achieving an early diagnosis effect on breast cancer. Therefore, the combined detection of the breast cancer biomarkers miRNA-21 and miRNA-155 is realized by constructing a genetic engineering microbial sensor, and the early diagnosis of the breast cancer can be realized.
disclosure of Invention
In order to overcome the problems in the prior art, the invention provides a microbial sensor for early detection of breast cancer, which comprises a detection chip of the microbial sensor and a detection system, wherein a gene circuit of a logical AND gate is formed by utilizing the action site characteristics of integrase FimE and integrase BxB1 in a gene recombination mode, a genetic engineering strain capable of realizing the combined detection of miRNA-21 and miRNA-155 is constructed, a micro-fluidic chip is prepared for carrying out the expanded culture of the microbial sensor, a portable fluorescence detection device is carried, and the simultaneous detection of two biomarkers is realized by detecting the fluorescence signal of fluorescent protein expressed by escherichia coli.
The invention provides the following technical scheme:
A microbial sensor for early detection of breast cancer is an engineered strain of genetic recombination, and the gene sequence of a recombinant plasmid introduced by the strain is as follows in sequence: a constitutive promoter, an RBS sequence, a sequence complementary to miRNA-155, a gene sequence for expressing tetR protein, a terminator, a first tetracycline promoter Ptet, an integrase FimE gene sequence and a terminator.
Further, the microbial sensor is used for detection of miRNA-155.
Further, the following gene sequences were also linked downstream of the gene sequences expressing tetR protein and terminator: a second tetracycline promoter, Ptet, a tara sequence, a terminator; the CrRNA sequence is also connected with the downstream of the first tetracycline promoter Ptet.
A microbial sensor for early detection of breast cancer is an engineered strain of genetic recombination, and the gene sequence of a recombinant plasmid introduced by the strain is as follows in sequence: a constitutive promoter, an RBS sequence, a sequence complementary to miRNA-21, a gene sequence for expressing lacI protein, a terminator, a first lactose promoter Plac, an integrase BxB1 gene sequence and a terminator.
Further, the microbial sensor is used for detection of miRNA-21.
Further, the following gene sequences are connected with the downstream of the gene sequences for expressing lacI protein and terminator: a second lactose promoter, Plac, TaRNA sequence, terminator; a CrRNA sequence was also ligated downstream of the first lactose promoter Plac.
A microbial sensor for early detection of breast cancer is an engineered strain of genetic recombination, and the gene sequence of a recombinant plasmid introduced by the strain is as follows in sequence: a constitutive promoter, an RBS sequence, a sequence complementary to miRNA-155, a gene sequence for expressing tetR protein, a terminator, a first tetracycline promoter Ptet, an integrase fimE gene sequence, a terminator, a constitutive promoter, an RBS sequence, a sequence complementary to miRNA-21, a gene sequence for expressing lacI protein, a terminator, a first lactose promoter Plac, an integrase BxB1 gene sequence, a terminator, a constitutive promoter, an integrase fimE action site gene sequence, an integrase BXB1 action site gene sequence, an RBS sequence, a fluorescent protein gene sequence and a terminator.
Further, the fluorescent protein is GFP.
Further, the following gene sequences were linked downstream of the gene sequences expressing tetR protein and terminator: a second tetracycline promoter, Ptet, a tara sequence, a terminator; a CrRNA sequence is also connected with the downstream of the first tetracycline promoter Ptet; the following gene sequences were ligated downstream of the gene sequence expressing lacI protein and terminator: a second lactose promoter, Plac, TaRNA sequence, terminator; a CrRNA sequence was also ligated downstream of the first lactose promoter Plac.
further, the microbial sensor is used for the combined detection of miRNA-155 and miRNA-21.
Further, the microbial sensor is made into a lyophilized powder.
Further, the engineering strain is escherichia coli.
A micro-fluidic detection chip containing a micro-biological sensor for early detection of breast cancer comprises a sample inlet, a sample outlet layer, a channel layer, a culture chamber layer and a substrate layer from top to bottom in sequence, wherein the layers are connected through a thermal bonding technology, and the sample inlet and the sample outlet layer are respectively provided with a sample inlet and a sample outlet on two sides of the chip; the culture cavity chamber layer is provided with a culture cavity chamber in the middle of the chip, and the microbial sensor is arranged in the culture cavity chamber layer; the channel layer is provided with channels which are respectively connected with the sample inlet and the culture chamber, and the sample outlet and the culture chamber in the chip.
Furthermore, the thickness of the sample inlet, the sample outlet layer, the channel layer and the substrate layer is 1-2 mm; the thickness of the culture chamber layer is 2-2.5 mm.
Further, the thermal bonding is carried out in a fiber van-type electric furnace, the temperature of the thermal bonding is 170-180 ℃, and the time of the thermal bonding is 18-25 min.
Furthermore, the microbial sensor is made into dry powder and fixed in the culture chamber.
The utility model provides a contain little biosensor detecting system who is used for early detection of breast cancer of detection chip, detecting system includes the ultraviolet source detect chip, control by temperature change heating module and imaging analysis module, detect the chip and load in control by temperature change heating module, the culture cavity of detection chip is shone to the ultraviolet source, and imaging analysis module shoots the culture cavity of detection chip to detect the fluorescence signal in the analysis photo.
Furthermore, the imaging analysis module is a smart phone, a rear camera of the smart phone is used for photographing and imaging, and a picture is displayed in a screen of the smart phone to perform detection and analysis on a fluorescence signal in the comparison sheet.
By adopting the technical scheme, the invention has the following beneficial effects:
1. According to the invention, through the characteristic that the integrase BxB1 and FimE are specifically combined with action sites thereof to generate conformational change to start fluorescent protein gene expression, a recombinant plasmid with a 'logic gate' gene circuit is constructed and is introduced into an escherichia coli engineering strain to form an escherichia coli microbial sensor, when miRNA-21 and miRNA-155 exist in a sample to be detected simultaneously, the gene circuit is started to express green fluorescent protein and emit a fluorescent signal, so that the early detection of breast cancer is realized, and the detection method is simple, sensitive and high in accuracy.
2. According to the design of the gene circuit, the stem-loop structure of the CrRNA is changed after the TaRNA and the CrRNA are combined, the inhibition effect of the CrRNA on the downstream gene expression is relieved, the structures of gene 'lock' and 'key' are added, the background fluorescence signal is effectively reduced, and the detection accuracy is improved.
3. The micro-fluidic detection chip fixes the microbial sensor in the detection chamber, controls the culture medium to flow in the culture chamber through the micro-fluidic technology, enlarges signals amplified by the cultured microbial sensor, and enhances the strength of the detection signals.
4. The microbial sensor detection system for early detection of breast cancer can utilize a smart phone to finish imaging and detection analysis of fluorescence signals, and the whole system is simple in structure and portable.
Drawings
FIG. 1 is a gene circuit layout diagram of a microbial sensor for detection of miRNA-155 in an embodiment of the invention;
FIG. 2 is a gene plasmid map of a microbial sensor for detection of miRNA-155 in an embodiment of the present invention;
FIG. 3 is a gene circuit layout diagram of a microbial sensor for detection of miRNA-21 in an embodiment of the invention;
FIG. 4 is a gene plasmid map of a microbial sensor for detection of miRNA-21 in an embodiment of the invention;
FIG. 5 is a gene circuit layout diagram of a microbial sensor for combined detection of miRNA-21 and miRNA-155 in an embodiment of the invention;
FIG. 6 is a schematic structural diagram of a microfluidic chip according to an embodiment of the present invention;
FIG. 7 is a schematic view showing the structure of a detection system of a microorganism sensor in the embodiment of the present invention.
The device comprises a sample inlet, a sample outlet layer, 11, a sample inlet, 12, a sample outlet, 2, a channel layer, 3, a culture chamber layer, 31, a culture chamber, 4 and a basal layer.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail with reference to the accompanying drawings and embodiments. It should be understood that the block diagrams and specific examples are set forth only for the purpose of illustrating the invention and are not to be construed as limiting the invention.
Example 1
As shown in figures 1 and 2, the invention provides a microbial sensor for early detection of breast cancer, the microbial sensor is a genetically recombinant engineering strain, and the gene sequence of a recombinant plasmid introduced by the strain is as follows: a constitutive promoter, an RBS sequence, a sequence complementary to miRNA-155, a gene sequence for expressing tetR protein, a terminator, a first tetracycline promoter Ptet, an integrase FimE gene sequence and a terminator. the tetR protein can be combined with a tetracycline promoter Ptet to inhibit the transcription and translation of genes downstream of the promoter Ptet. When miRNA-155 does not exist in the object to be detected, tetR protein is expressed to inhibit the expression of downstream genes of a promoter Ptet, so that the expression of integrase fimE is also inhibited; when miRNA-155 exists in the object to be detected, the miRNA-155 is combined with the specific complementary sequence, the expression of the tetR protein is inhibited, the promoter Ptet is not inhibited, and the integrase fimE is expressed. The integrase FimE can be detected by conventional detection methods, such as ELISA and the like. The microbial sensor can be used for detection of miRNA-155.
When miRNA does not exist in a sample, the initiation of integrase is inhibited, but a small amount of integrase is still expressed, in order to reduce background signals, the structures of gene 'Lock' and 'Key', namely CrRNA (Lock) and TaRNA (Key), are added in the design of a gene circuit, and when the expression level of the TaRNA is low, the CrRNA inhibits the expression of downstream integrase genes; when the expression quantity of the TaRNA is increased, the TaRNA interacts with CrRNA to relieve inhibition, and integrase genes are expressed, so that the background signal can be greatly reduced by the group of gene structures.
Therefore, the following gene sequences were also linked downstream of the gene sequence expressing the tetR protein and the terminator: a second tetracycline promoter, Ptet, a tara sequence, a terminator; the CrRNA sequence is also connected with the downstream of the first tetracycline promoter Ptet.
Example 2
as shown in fig. 3 and 4, the invention provides a microbial sensor for early detection of breast cancer, the microbial sensor is a genetically recombinant engineering strain, and the gene sequence of a recombinant plasmid introduced by the strain is as follows: a constitutive promoter, an RBS sequence, a sequence complementary to miRNA-21, a gene sequence for expressing lacI protein, a terminator, a first lactose promoter Plac, an integrase BxB1 gene sequence and a terminator.
The lacI protein can be combined with a promoter Plac to inhibit the transcription and translation of a downstream gene of the promoter Plac, and when miRNA-21 does not exist in a sample to be tested, the lacI protein is expressed to inhibit the expression of the downstream gene of the promoter Plac, so that the expression of integrase BxB1 is also inhibited; when miRNA-21 exists in the test object, the miRNA-21 is combined with the specific complementary sequence, the expression of lacI protein is inhibited, the promoter Plac is not inhibited, and the integrase BxB1 is expressed to be used for detecting the miRNA-21. The integrase BxB1 can be detected by conventional detection methods, such as ELISA and the like. The microbial sensor can be used for detection of miRNA-21.
In order to reduce the background signal, the following gene sequences were also ligated downstream of the gene sequences of the lacI protein and terminator: a second lactose promoter, Plac, TaRNA sequence, terminator; a CrRNA sequence was also ligated downstream of the first lactose promoter, Plac.
Example 3
The invention utilizes the action site characteristics of integrase FimE and integrase BxB1 to form a gene circuit of a logic AND gate, a promoter Ptet controls the expression of the integrase FimE, a promoter Plac controls the expression of the integrase BxB1, and only when the integrase FimE and the integrase BxB1 exist simultaneously, the corresponding action site conformation can be changed, thereby starting the expression of a downstream fluorescent gene. The recombinant plasmid is introduced into engineering strains such as escherichia coli, when miRNA-21 and miRNA-155 exist in a sample to be detected at the same time, a gene circuit is expressed, and the escherichia coli expresses fluorescent protein.
As shown in FIG. 5, the invention provides a microbial sensor for early detection of breast cancer, the microbial sensor is a genetically recombinant engineered strain, and the gene sequences of recombinant plasmids introduced by the strain are as follows: a constitutive promoter, an RBS sequence, a sequence complementary to miRNA-155, a gene sequence for expressing tetR protein, a terminator, a first tetracycline promoter Ptet, an integrase fimE gene sequence, a terminator, a constitutive promoter, an RBS sequence, a sequence complementary to miRNA-21, a gene sequence for expressing lacI protein, a terminator, a first lactose promoter Plac, an integrase BxB1 gene sequence, a terminator, a constitutive promoter, an integrase fimE action site gene sequence, an integrase BXB1 action site gene sequence, an RBS sequence, a fluorescent protein gene sequence and a terminator. Wherein the fluorescent protein may be GFP.
to reduce background signal, the following gene sequences were ligated downstream of the gene sequence expressing the tetR protein and the terminator: a second tetracycline promoter, Ptet, a tara sequence, a terminator; a CrRNA sequence is also connected with the downstream of the first tetracycline promoter Ptet; the following gene sequences were ligated downstream of the gene sequence expressing the lacI protein and terminator: a second lactose promoter, Plac, TaRNA sequence, terminator; a CrRNA sequence was also ligated downstream of the first lactose promoter, Plac.
Only when miRNA-155 and miRNA-21 exist in the object to be detected at the same time, the two integrases are expressed and are respectively combined with action sites thereof, so that the expression of downstream green fluorescent protein genes is excited; when the target miRNA does not exist or only exists in the object to be detected, the conformation of the action site of the integrase cannot be changed simultaneously, the downstream green fluorescent protein gene is not expressed, and the detection system does not generate a fluorescent signal. Thus, the microbial sensor in this example was used for the combined detection of miRNA-155 and miRNA-21.
Example 4
As shown in FIG. 6, the micro-fluidic detection chip comprising the above-mentioned biosensor for early detection of breast cancer comprises, from top to bottom, a sample inlet, a sample outlet layer 1, a channel layer 2, a culture chamber layer 3 and a substrate layer 4, which are connected by thermal bonding technology. The sample inlet and the sample outlet layer are respectively provided with a sample inlet 11 and a sample outlet 12 at two sides of the chip. The culture chamber layer is provided with a culture chamber 31 in the middle of the chip, and the microbial sensor is placed in the culture chamber, preferably, the microbial sensor is made into dry powder and fixed in the culture chamber. The channel layer is provided with channels which are respectively connected with the sample inlet and the culture chamber, and the sample outlet and the culture chamber in the chip.
Wherein the thickness of the sample inlet, the sample outlet layer, the channel layer and the substrate layer is 1-2 mm; the thickness of the culture chamber layer is 2-2.5 mm. The thermal bonding is carried out in a fiber box type electric furnace, the temperature of the thermal bonding is 170-180 ℃, and the time of the thermal bonding is 18-25 min.
After the chip is manufactured, the overnight cultured microbial sensor bacterial liquid can be injected into the microfluidic chip culture chamber and placed in a vacuum freeze dryer for freeze drying. After the freeze-drying is finished, the microbial sensor can be fixed in the culture chamber in the form of dry powder. During detection, a sample to be detected and an LB culture medium are injected into a culture chamber, and the bacteria liquid dry powder is subjected to resuscitation culture, so that the detection of miRNA in the sample can be completed.
example 5
the invention provides a microbial sensor detection system containing the detection chip and used for early detection of breast cancer, as shown in fig. 7, the detection system comprises an ultraviolet light source, a detection chip, a temperature control heating module and an imaging analysis module, the detection chip is loaded in the temperature control heating module, the ultraviolet light source irradiates a culture chamber of the detection chip, and the imaging analysis module takes a picture of the culture chamber of the detection chip and detects and analyzes a fluorescent signal in the picture. The exciting light can be high-power LED ultraviolet lamp (365nm) as exciting light source, and 365nm +/-10 nm narrow-band filter is placed at the position of the light source.
The temperature control heating module comprises a bearing bottom plate and a chip loading clamping strip, wherein an aluminum oxide ceramic plate is arranged in the bearing bottom plate, and the temperature of the chip during culture is controlled by a connecting circuit. The chip loading strip has the function of fixing the microfluidic chip and placing the chip on the upper part of the bearing bottom plate.
The imaging analysis module sets up the dichroic mirror that can filter out the below 509nm optical signal of wavelength in fluorescence signal collection department, can prevent that background ultraviolet from passing through, improves the SNR. Preferably, the imaging analysis module can be a smart phone, a rear camera of the smart phone is used for photographing and imaging, a picture is displayed in a screen of the smart phone, and detection and analysis are performed on a fluorescence signal in the film. And the smart phone is communicated with a circuit of the system through a Bluetooth connection function, and then the temperature is set through a switch of a control circuit of the smart phone and a temperature control heating module.
The above-mentioned embodiments only express the embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (18)
1. A microbial sensor for early detection of breast cancer is characterized in that the microbial sensor is a genetically recombinant engineering strain, and a recombinant plasmid gene sequence introduced by the strain is as follows in sequence: a constitutive promoter, an RBS sequence, a sequence complementary to miRNA-155, a gene sequence for expressing tetR protein, a terminator, a first tetracycline promoter Ptet, an integrase FimE gene sequence and a terminator.
2. The microbial sensor for early detection of breast cancer according to claim 1, wherein the microbial sensor is used for detection against miRNA-155.
3. The biosensor for the early detection of breast cancer according to claim 1, wherein the following gene sequences are further ligated downstream of the gene sequences expressing tetR protein and terminator: a second tetracycline promoter, Ptet, a tara sequence, a terminator; the CrRNA sequence is also connected with the downstream of the first tetracycline promoter Ptet.
4. A microbial sensor for early detection of breast cancer is characterized in that the microbial sensor is a genetically recombinant engineering strain, and a recombinant plasmid gene sequence introduced by the strain is as follows in sequence: a constitutive promoter, an RBS sequence, a sequence complementary to miRNA-21, a gene sequence for expressing lacI protein, a terminator, a first lactose promoter Plac, an integrase BxB1 gene sequence and a terminator.
5. The microbial sensor for early detection of breast cancer according to claim 4, wherein the microbial sensor is used for detection of miRNA-21.
6. The biosensor for the early detection of breast cancer according to claim 4, wherein the following gene sequences are further ligated downstream of the gene sequence expressing lacI protein and terminator: a second lactose promoter, Plac, TaRNA sequence, terminator; a CrRNA sequence was also ligated downstream of the first lactose promoter Plac.
7. A microbial sensor for early detection of breast cancer is characterized in that the microbial sensor is a genetically recombinant engineering strain, and a recombinant plasmid gene sequence introduced by the strain is as follows in sequence: a constitutive promoter, an RBS sequence, a sequence complementary to miRNA-155, a gene sequence for expressing tetR protein, a terminator, a first tetracycline promoter Ptet, an integrase fimE gene sequence, a terminator, a constitutive promoter, an RBS sequence, a sequence complementary to miRNA-21, a gene sequence for expressing lacI protein, a terminator, a first lactose promoter Plac, an integrase BxB1 gene sequence, a terminator, a constitutive promoter, an integrase fimE action site gene sequence, an integrase BXB1 action site gene sequence, an RBS sequence, a fluorescent protein gene sequence and a terminator.
8. The biosensor in accordance with claim 7, wherein said fluorescent protein is GFP.
9. The biosensor for the early detection of breast cancer according to claim 7, wherein the following gene sequences are linked downstream of the gene sequences expressing tetR protein and terminator: a second tetracycline promoter, Ptet, a tara sequence, a terminator; a CrRNA sequence is also connected with the downstream of the first tetracycline promoter Ptet; the following gene sequences were ligated downstream of the gene sequence expressing lacI protein and terminator: a second lactose promoter, Plac, TaRNA sequence, terminator; a CrRNA sequence was also ligated downstream of the first lactose promoter Plac.
10. The microbial sensor for early detection of breast cancer according to claim 7, wherein the microbial sensor is used for combined detection of miRNA-155 and miRNA-21.
11. The microbial sensor for the early detection of breast cancer according to any one of claims 1 to 10, wherein the microbial sensor is prepared as a lyophilized powder.
12. the biosensor for the early detection of breast cancer according to any one of claims 1 to 10, wherein the engineered strain is Escherichia coli.
13. a microfluidic detection chip comprising the micro-biological sensor for early detection of breast cancer according to any one of claims 1 to 10, wherein the microfluidic detection chip comprises a sample inlet, a sample outlet layer, a channel layer, a culture chamber layer and a substrate layer in sequence from top to bottom, the layers are connected by thermal bonding technology, and the sample inlet and the sample outlet layer are respectively provided with a sample inlet and a sample outlet on two sides of the chip; the culture cavity chamber layer is provided with a culture cavity chamber in the middle of the chip, and the microbial sensor is arranged in the culture cavity chamber layer; the channel layer is provided with channels which are respectively connected with the sample inlet and the culture chamber, and the sample outlet and the culture chamber in the chip.
14. The microfluidic detection chip according to claim 13, wherein the thickness of the sample inlet, the sample outlet layer, the channel layer and the substrate layer is 1-2 mm; the thickness of the culture chamber layer is 2-2.5 mm.
15. The microfluidic detection chip according to claim 13, wherein the thermal bonding is performed in a fiber van-type electric furnace, the temperature of the thermal bonding is 170-180 ℃, and the thermal bonding time is 18-25 min.
16. The detecting chip according to claim 13, wherein the biosensor is fixed in the culture chamber as a dry powder.
17. The system for detecting the micro-organism sensor for the early detection of the breast cancer, which comprises the detection chip of claim 13, is characterized in that the detection system comprises an ultraviolet light source, the detection chip, a temperature control heating module and an imaging analysis module, wherein the detection chip is loaded in the temperature control heating module, the ultraviolet light source irradiates a culture chamber of the detection chip, and the imaging analysis module takes a picture of the culture chamber of the detection chip and detects and analyzes a fluorescent signal in the picture.
18. The detection system according to claim 17, wherein the imaging analysis module is a smart phone, a rear camera of the smart phone is used for photographing and imaging, and a picture is displayed in a screen of the smart phone to perform detection analysis on the fluorescence signal in the comparison sheet.
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