CN111307770A - PCR detection device and method - Google Patents
PCR detection device and method Download PDFInfo
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- CN111307770A CN111307770A CN202010118084.4A CN202010118084A CN111307770A CN 111307770 A CN111307770 A CN 111307770A CN 202010118084 A CN202010118084 A CN 202010118084A CN 111307770 A CN111307770 A CN 111307770A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
Abstract
The invention provides a PCR detection device and a method, which relate to the technical field of biological detection, wherein the PCR detection device comprises an excitation end, a detection end and a conversion module; the excitation end comprises M excitation optical fiber bundles, and each excitation optical fiber bundle is provided with N optical fibers; the detection end comprises M detection optical fiber bundles, and each detection optical fiber bundle is provided with N optical fibers; wherein M is more than or equal to 2, and N is more than or equal to 2; the conversion module converts one of the M excitation optical fiber bundles and one of the M detection optical fiber bundles into N groups of sample side optical fibers; each N sets of sample side fibers correspond to a column of sample wells. When the high-flux detection is carried out, the whole optical fiber arrangement is simple and clear through the optical fiber arrangement mode, the whole volume and complexity are reduced, the occupied space is small, and the structure is simplified. And when a large amount of samples are detected in a high-flux manner, scanning is not needed, and the sample detection time is shortened. In particular, the effect is more remarkable when the number of sample holes is larger and the number of optical fibers is larger.
Description
Technical Field
The invention relates to the technical field of biological detection, in particular to a PCR detection device and a method.
Background
Real-time fluorescence quantitative Polymerase Chain Reaction (PCR) is a quantitative detection method, and can be used for labeling and tracking a PCR product through a fluorescent dye or a fluorescence-labeled specific probe, monitoring a Reaction process in real time on line, and analyzing the product. When the fluorescence quantitative detection is carried out on the PCR, the conventional PCR light path is transmitted through the dichroic mirror and the reflecting mirror, which can cause the problem of optical path difference of different reaction areas, and the problem can be solved by utilizing optical fiber transmission.
In the existing multi-fiber fluorescent signal detection device, an emission light path of a detection end and an excitation light path of an excitation end respectively use two independent fiber bundles, and fiber arrays of the emission fiber bundles correspond to sample holes one by one. Thus, in high-throughput detection (a large number of sample wells), the number of optical fibers used is also large, for example, 96 optical fibers are required for an 8 × 12-standard PCR well plate, both the excitation optical fiber for receiving the excitation light of the light source and the detection optical fiber for receiving the fluorescence for detection, which results in a complicated arrangement of the optical fibers and a large space occupation.
Disclosure of Invention
The invention aims to provide a PCR detection device and a method thereof, which are used for solving the technical problems that the whole optical fiber arrangement is complicated and a huge space is occupied due to the large number of optical fibers of a multi-optical fiber fluorescent signal detection device.
The invention also aims to provide a PCR detection device, which solves the technical problems that in the existing multi-optical fiber fluorescence signal detection device, the volume of the excitation end of a plurality of optical fibers and the volume of the detection end of a plurality of optical fibers after the optical fibers are converged are too large, so that the light intensity received by the inner optical fiber and the outer optical fiber is not uniform and has difference after the excitation light of a light source irradiates the excitation end or the emission light irradiates the detection end. In addition, the technical problem that the detection result aiming at different reaction regions is inaccurate due to the fact that the positions of the optical fibers are far away from each other, detection errors of detection equipment at a detection end are further relieved, and when the optical fibers are converged, each optical fiber needs to be converged in a one-to-one correspondence mode, so that processing is difficult, cost is high, a large number of optical fibers are converged together, and when the number of the optical fibers is large, the positions corresponding to the optical fibers are not easy to find is solved.
In a first aspect, the PCR detection apparatus provided by the present invention comprises an excitation end, a detection end and a conversion module; the excitation end comprises M excitation optical fiber bundles, and each excitation optical fiber bundle is provided with N excitation optical fibers; the detection end comprises M detection optical fiber bundles, and each detection optical fiber bundle is provided with N detection optical fibers; wherein M is more than or equal to 2, and N is more than or equal to 2;
the conversion module converts one of the M excitation fiber bundles and one of the M detection fiber bundles into a set of sample-side fibers; each N sets of sample side fibers correspond to a column of sample wells.
With reference to the first aspect, the present invention provides a first possible implementation manner of the first aspect, wherein M excitation optical fiber bundles are transmitted at the excitation end for a distance in a manner of an excitation optical cable, and M detection optical fiber bundles are transmitted at the detection end for a distance in a manner of a detection optical cable.
With reference to the first aspect, the present invention provides a second possible implementation manner of the first aspect, wherein the conversion modules include M, each conversion module converts one excitation fiber bundle and one detection fiber bundle into N sets of sample-side optical fibers, and each sample hole corresponds to one set of sample-side optical fibers.
With reference to the first aspect, an embodiment of the present invention provides a third possible implementation manner of the first aspect, where the optical fiber module further includes an optical fiber holder, and the M conversion modules are placed on the optical fiber holder side by side.
With reference to the first aspect, the present invention provides a fourth possible implementation manner of the first aspect, wherein the M conversion modules are divided into multiple groups, and each group is placed on the fiber fixing frame side by side.
With reference to the first aspect, the present invention provides a fifth possible implementation manner of the first aspect, wherein the apparatus further comprises a sample-side installation module; the sample side optical fiber is installed on the installation module.
With reference to the first aspect, the present disclosure provides a sixth possible implementation manner of the first aspect, wherein the mounting module includes M mounting seats arranged side by side, and every N groups of the sample-side optical fibers are arranged on one of the mounting seats.
With reference to the first aspect, the present invention provides a seventh possible implementation manner of the first aspect, wherein the excitation terminal and the detection terminal are separately encapsulated in different shells.
With reference to the first aspect, the present invention provides an eighth possible implementation manner of the first aspect, wherein M excitation fiber bundles or M detection fiber bundles have corresponding numbers respectively.
In a second aspect, the present invention provides a PCR detection method applied to the PCR detection apparatus, the method including:
exciting light emitted by the exciting end is transmitted to the conversion module through the M exciting optical fiber bundles and then transmitted to the sample hole through the sample side optical fiber, and a sample to be detected is excited to obtain emitted light; wherein each excitation optical fiber bundle is provided with N excitation optical fibers, M is more than or equal to 2, and N is more than or equal to 2;
the emitted light is transmitted to the conversion module through the sample side optical fiber and then transmitted to the detector through M detection optical fiber bundles; each detection optical fiber bundle is provided with N detection optical fibers;
the detector detects the intensity of the fluorescent signal of the emitted light. The PCR detection device provided by the invention is respectively arranged at the excitation end and the detection end in a mode of M optical fiber bundles, each optical fiber bundle comprises N optical fibers, one of the M excitation optical fiber bundles and one of the M detection optical fiber bundles are converted into N groups of sample side optical fiber bundles through a conversion module, and each N groups of sample side optical fibers correspond to one row of sample holes. When the high-throughput detection is carried out on a large number of sample holes (such as 24, 48, 96, 192, 384 or more sample holes) and a large number of used optical fibers, the whole optical fiber arrangement is simple and clear through the optical fiber arrangement mode, the whole volume and complexity of the detection device are reduced, the occupied space is small, and the structure is simplified. And when a large amount of samples are detected in a high-flux manner, scanning is not needed, and the sample detection time is shortened. In particular, the effect is more remarkable when the number of sample holes is larger and the number of optical fibers is larger.
In addition, the PCR detection device provided by the invention also has the following beneficial effects:
(1) the M excitation optical fiber bundles and the M detection optical fiber bundles are transmitted in a mode of exciting the optical cable and detecting the optical cable respectively, the M X N excitation optical fibers can be gathered together at the exciting end and the M X N detection optical fibers can be gathered together at the detecting end instead of being diverged after coming out from the port, and therefore the size and the arrangement complexity of the whole optical fibers can be greatly reduced.
(2) According to the invention, as the prior art does not need to arrange a sleeve for each optical fiber, the processing and the manufacturing are more convenient, the volumes of the excitation end and the detection end and the cross-sectional areas on the two end faces can be reduced, after the cross-sectional areas are reduced, when the excitation light emitted from the light source enters the excitation optical fibers in the excitation optical cable, the uniformity of light intensity among the optical fibers is better, and the difference is small; the detection optical fiber in the detection optical cable can also reduce the detection error caused by the position of different optical fibers, thereby improving the detection accuracy and sensitivity.
(3) The conversion of the optical fiber bundle for detecting the optical fiber bundle is completed through one conversion module, and the conversion of N optical fibers can be realized because one optical fiber bundle comprises N optical fibers, so that a plurality of conversion structures are not needed, and the structure is further simplified.
(4) In this embodiment, the sample side optical fibers are arranged in groups, so that each mounting seat or each row of sample holes corresponds to each group of sample side optical fibers, and the position of a certain sample side optical fiber is easy to find; in addition, when a certain sample side optical fiber is damaged, the optical fiber is easy to replace, and the structural complexity and the cost can be reduced.
(5) An excitation light path where an excitation end is located and an emission light path where a detection end is located are independently arranged, and the excitation end and the detection end are specifically arranged in different shells, so that a certain light path can be independently replaced and maintained when a fault occurs;
(6) the M excitation optical fiber bundles or the M detection optical fiber bundles are respectively provided with corresponding numbers, so that the positions of the excitation optical fibers or the detection optical fibers are easy to search; in addition, the optical fiber can be easily replaced when the excitation optical fiber or the detection optical fiber is damaged. According to the PCR detection method provided by the invention, excitation light is transmitted through M excitation optical fiber bundles at an excitation end, each excitation optical fiber bundle comprises N excitation optical fibers, emission light is transmitted through M detection optical fiber bundles at a detection end, and each detection optical fiber bundle comprises N detection optical fibers. High-throughput detection can be rapidly achieved through a large number of optical fibers. In particular, the effect is more remarkable when the number of sample holes is larger and the number of optical fibers is larger.
Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. The objectives and other advantages of the invention will be realized and attained by the structure particularly pointed out in the written description and claims hereof as well as the appended drawings.
In order to make the aforementioned and other objects, features and advantages of the present invention comprehensible, preferred embodiments accompanied with figures are described in detail below.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a schematic diagram of a PCR detection apparatus provided in an embodiment of the present invention;
fig. 2 (a) and (b) are schematic diagrams of the optical fiber arrangement of the excitation end provided by the embodiment of the present invention;
fig. 3 (a) and (b) are schematic diagrams of the optical fiber arrangement of the detection end provided by the embodiment of the invention;
FIG. 4 is a schematic diagram of an excitation end fiber bundle and a detection end fiber bundle converted into sample side fibers according to an embodiment of the present invention;
FIG. 5 is a schematic diagram of a sample side optical fiber provided by an embodiment of the present invention;
FIG. 6 is a schematic view of a fiber holder according to an embodiment of the present invention;
FIG. 7 is a schematic view of another fiber holder according to an embodiment of the present invention;
FIG. 8 is a schematic view of a sample side fiber mount provided by an embodiment of the present invention;
fig. 9 is a schematic diagram of a sample well provided by an embodiment of the present invention.
Icon: 1-a light source; 2-a convex lens; 3-excitation light filter wheel; 4-an excitation end; 5-an excitation optical cable; 6-a detector; 7-a receiving optical filter; 8-detection end; 9-a lens group; 10-detecting the optical cable; 11-a conversion module; 12-optical fiber fixing frame.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments, and it should be understood that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the existing multi-fiber fluorescent signal detection device, an emission light path of a detection end and an excitation light path of an excitation end respectively use two independent fiber bundles, and fiber arrays of the emission fiber bundles correspond to sample holes one by one. Thus, in high-throughput detection (a large number of sample wells), the number of optical fibers used is also large, for example, 96 optical fibers are required for an 8 × 12-standard PCR well plate, both the excitation optical fiber for receiving the excitation light of the light source and the detection optical fiber for receiving the fluorescence for detection, which results in a complicated arrangement of the optical fibers and a large space occupation. Based on this, the PCR detection device provided by the embodiment of the invention can enable the whole optical fiber arrangement to be simple and clear, reduce the whole volume and complexity of the detection device, occupy small space and simplify the structure during high-throughput detection.
For the understanding of the present embodiment, a detailed description will be given to a PCR detection apparatus disclosed in the present embodiment.
The PCR detection device provided by the embodiment of the invention comprises: the device comprises an excitation end, a detection end and a conversion module; the excitation end comprises M excitation optical fiber bundles, and each excitation optical fiber bundle is provided with N excitation optical fibers; the detection end comprises M detection optical fiber bundles, and each detection optical fiber bundle is provided with N detection optical fibers; wherein M is more than or equal to 2, and N is more than or equal to 2; the conversion module converts one of the M excitation optical fiber bundles and one of the M detection optical fiber bundles into N groups of sample side optical fibers, each N groups of sample side optical fibers correspond to one row of sample holes, and M rows of sample holes are shared.
Specifically, the excitation end emits excitation light, the excitation light is transmitted to the sample side optical fiber through the excitation optical fiber bundle, the sample to be detected on the sample side is excited through the sample hole to generate emission light, the emission light enters the sample side optical fiber and is projected into the detector through the detection optical fiber bundle, and the intensity of a fluorescence signal of the emission light is detected through the detector due to the fact that the emission light has the fluorescence signal.
In this embodiment, the excitation end emits excitation light through M excitation fiber bundles, each excitation fiber bundle is provided with N fibers, and then M × N excitation fibers and M × N detection fibers are converted by the conversion module to become M × N groups of sample side fibers capable of transmitting excitation light and receiving fluorescence, wherein one excitation fiber and one detection fiber can be converted to a sample side fiber, or bundled together to a bundle of sample side fibers, or converted to two sample side fibers arranged in parallel, and therefore, each group of sample side fibers can be one or a bundle of sample side fibers, or two sample side fibers arranged in parallel, one of the sample side fibers is used for transmitting excitation light, and the other is used for receiving fluorescence, of course, the two sample side fibers may be arranged in any manner, as long as they are aligned with one sample well at the same time.
It should be noted that specific values of M and N can be set according to the specification of the PCR well plate carrying the sample, and the number of sample wells on the PCR well plate is not limited, for example, 12 wells, 24 wells, 48 wells, 96 wells, 192 wells, 384 wells, and the like. The arrangement of the sample wells is not limited, and for example, 96 wells may have the specification of 8 × 12, 12 × 8, 16 × 6, 6 × 16, and the like. For example, for a 12 × 8 PCR well plate, 96 excitation fibers for receiving excitation light from the light source and 96 detection fibers for receiving fluorescence for detection are required, and in this case, M may be 12 and N may be 8.
According to the PCR detection device provided by the embodiment of the invention, the excitation end and the detection end are respectively arranged in a mode of M optical fiber bundles, each optical fiber bundle comprises N optical fibers, one of the M excitation optical fiber bundles and one of the M detection optical fiber bundles are converted into N groups of sample side optical fiber bundles through the conversion module, and each N groups of sample side optical fibers correspond to one row of sample holes. When the high-throughput detection with more sample holes and large used optical fibers is carried out, the whole optical fibers can be simply and clearly arranged by the optical fiber arrangement mode, the whole volume and complexity of the detection device are reduced, the occupied space is small, and the structure is simplified. And when a large amount of samples are detected in a high-flux manner, scanning is not needed, and the sample detection time is shortened. In particular, the effect is more remarkable when the number of sample holes is larger and the number of optical fibers is larger.
In an alternative embodiment, the M excitation fiber bundles are transported over a distance at the excitation end by means of an excitation optical cable and the M detection fiber bundles are transported over a distance at the detection end by means of a detection optical cable. Because M exciting optical fiber bundles comprise M exciting optical fibers, and M detecting optical fiber bundles comprise M detecting optical fibers, the M exciting optical fibers and the M detecting optical fibers can be converged together at the exciting end and converged together at the detecting end instead of diverging from the port in a transmission mode of the exciting optical fiber bundle and the detecting optical fiber bundle, and therefore the volume and the arrangement complexity of the whole optical fiber can be greatly reduced.
FIG. 1 is a schematic diagram of a PCR detection apparatus according to an embodiment of the present invention.
As shown in fig. 1, a PCR detection apparatus provided in an embodiment of the present invention includes an excitation light path and an emission light path, where the excitation light path includes a light source 1, a convex lens 2, an excitation light filter wheel 3, an excitation end 4, and an excitation optical fiber disposed in the manner of an excitation optical cable 5, and the emission light path includes a detector 6, a receiving light filter 7, a detection end 8, and a detection optical fiber disposed in the manner of a detection optical cable 10.
Excitation light emitted by the light source 1 is focused by the convex lens 2 and then filtered by the excitation light filter wheel 3 to generate excitation light with a specific wavelength, the excitation light is transmitted through the excitation optical cable 5 coming out of the excitation end 4, M excitation optical fiber bundles are arranged in the excitation optical cable 5, and each excitation optical fiber bundle comprises N excitation optical fibers. M detection optical fiber bundles are arranged in a detection optical cable 10 from the detection end 8, and each detection optical fiber bundle comprises N detection optical fibers. The light source may be an LED or a laser, and may be a single light source or a plurality of light sources with different wavelengths.
The optical fiber of the excitation end 4 and the optical fiber of the detection end 8 are respectively dispersed after being transmitted for a distance in the manner of the excitation optical cable 5 and the detection optical cable 10, and then are converted by a conversion module, specifically, the conversion can be performed at the block shown in fig. 1. The purpose of the conversion is to bring together one excitation fiber in the excitation optical cable 5 and one detection fiber in the detection optical cable 10 into a set of sample side fibers which are applied at the sample hole.
In practical application, the excitation light is transmitted for a distance through the excitation optical cable 5 and enters the sample-side optical fiber, and then is converged by the lens group 9 and emitted to the sample hole to excite the sample to be detected to generate emission light with a fluorescence signal, the emission light enters the sample-side optical fiber, is projected into the detector 6 at the detection end through the detection optical cable 10, the fluorescence signal intensity of the emission light is detected by the detector, and in addition, the emission light can be filtered by the receiving optical filter 7 before entering the detector.
Specifically, the excitation fibers may be arranged at the excitation end 4 in a manner shown in (a) or (b) in fig. 2, and the detection fibers may be arranged at the detection end 8 in a manner shown in (a) or (b) in fig. 3, wherein the excitation fibers are converged at the excitation end in a form of a plurality of excitation fiber bundles to form the excitation optical cable 5, and the detection fibers are converged at the detection end in a form of a plurality of detection fiber bundles to form the detection optical cable 10. Taking 96 sample wells as an example, there are 12 excitation fiber bundles A, B … in the excitation optical cable 5, and each excitation fiber bundle contains 8 excitation fibers a1 … a8, B1 … B8, and the like. The detection optical cable also comprises 12 detection optical fiber bundles A 'and B' …, and each detection optical fiber bundle comprises 8 detection optical fibers A '1 … A' 8, B '1 … B' 8 and the like.
It should be noted that each excitation fiber bundle and each detection fiber bundle may be coated with a coating layer, or may not have a coating layer. As shown in fig. 2 (a) and fig. 3 (a), the 12 excitation optical fiber bundles and the 12 detection optical fiber bundles are coated by an outer coating layer; as shown in fig. 2 (b) and 3 (b), 12 optical fiber bundles may be formed by combining 8 optical fibers into one optical fiber bundle, and the 12 optical fiber bundles may be transmitted in the form of an optical cable, without a cladding layer.
Through the arrangement mode, 96 excitation optical fibers of the excitation end are divided into 12 excitation optical fiber bundles, each excitation optical fiber bundle comprises 8 excitation optical fibers, and after the 8 excitation optical fibers are converged in each excitation optical fiber bundle, the 12 excitation optical fiber bundles are converged together; similarly, 96 detection optical fibers at the detection end are divided into 12 detection optical fiber bundles, each detection optical fiber bundle comprises 8 detection optical fibers, and after 8 excitation optical fibers are converged in each excitation optical fiber bundle, the 12 excitation optical fiber bundles are converged together.
Therefore, the embodiment of the invention does not need to configure a sleeve for each optical fiber as the prior art, so the processing and the manufacturing are more convenient, the volumes of the excitation end and the detection end and the cross-sectional areas on the two end surfaces can be reduced, after the cross-sectional areas are reduced, when the excitation light emitted from the light source enters the excitation optical fiber in the excitation optical cable, the light intensity uniformity among the optical fibers is better, and the difference is small; the detection optical fiber in the detection optical cable can also reduce the detection error caused by the position of different optical fibers, thereby improving the detection accuracy and sensitivity.
In an alternative embodiment, the conversion module includes M, each conversion module converts one excitation fiber bundle and one detection fiber bundle into N sets of sample side fibers, and each sample well corresponds to one set of sample side fibers. The number of the conversion modules is the same as that of the excitation optical fiber bundles and the detection optical fiber bundles, the conversion of one optical fiber bundle detection optical fiber bundle is completed only through one conversion module, and the conversion of N optical fibers can be realized because one optical fiber bundle comprises N optical fibers, so that a plurality of conversion structures are not needed, and the structure is further simplified.
As shown in fig. 4, the conversion process of the fiber bundle is: the excitation fiber a1 in the excitation fiber bundle a and the detection fiber a ' 1 in the detection fiber bundle a ' are converted into a group of sample side fibers a1 applied to the sample hole through the conversion module 11, and the excitation fiber a2 and the detection fiber a ' 2 are converted into a group of sample side fibers a2 applied to the sample hole through the conversion module 11, and A3, a4, a5, a6, a7 and A8 are obtained in the same way. Therefore, 8 excitation fibers in one excitation fiber bundle a and 8 detection fibers in one detection fiber bundle a' are converted into 8 sets of sample-side fibers a1, a2, a3, a4, a5, a6, a7, and a 8. The 8 groups of sample side optical fibers may be 8 or 8 bundles of sample side optical fibers, or each group may include two sample side optical fibers, and the two sample side optical fibers of each group may be arranged together in any manner as long as they can be simultaneously aligned with one sample hole. The 8 sample side optical fibers a1, a2, a3, a4, a5, a6, a7 and a8 are used for integrating the functions of light source excitation and fluorescence detection, exciting and detecting in the same direction, and selecting inner ring excitation or outer ring excitation; as shown in fig. 5, the excitation fiber a1 and the detection fiber a '1 may be coupled into one sample-side fiber a1, and 8 excitation fibers in the excitation fiber bundle a and 8 detection fibers in the detection fiber bundle a' are coupled to obtain 8 sample-side fibers a1, a2, a3, a4, a5, a6, a7, and A8, where the 8 sample-side fibers may be coupled to perform inner ring excitation or outer ring excitation.
The 8 bundles of sample-side optical fibers a1, a2, A3, a4, a5, a6, a7, and a8 mentioned above may be obtained by bundling the excitation optical fiber a1 and the detection optical fiber a '1 to form a bundle of sample-side optical fibers a1, and bundling the excitation optical fiber a2 and the detection optical fiber a' 2 to form a bundle of sample-side optical fibers a2, similarly obtaining A3, a4, a5, a6, a7, and a 8. After conversion in the above manner, only one set of sample side optical fibers can be arranged at each sample hole, excitation light is transmitted to the sample through the sample side optical fibers, and emitted light with a fluorescent signal is received, so that the number of the optical fibers at the sample side can be reduced, and the structure is simplified.
In an alternative embodiment, as shown in fig. 6, the PCR detection apparatus according to the embodiment of the present invention further includes a fiber holder 12, and the M conversion modules 11 are placed on the fiber holder 12 side by side. Specifically, M holes may be formed in the optical fiber holder 12, and one conversion module is disposed in each hole, so that the optical fiber arrangement is neater.
To further reduce the volume occupied by the optical fibers, as shown in fig. 7, the M conversion modules 11 may be divided into a plurality of groups, each group being placed side by side on the fiber holder 12. For example, for 96 excitation fibers of 12 excitation fiber bundles, 12 conversion modules are required in total, the 12 conversion modules can be arranged on a fiber fixing frame in two groups side by side, each group of 6 conversion modules can be arranged in holes of the fiber fixing frame, and therefore the arrangement of the fibers is tidier, the space is fully utilized, and the volume occupied by the fibers is reduced. Of course, the grouping method is not fixed, and may be not only 2 rows by 6, but also 3 rows by 4, and the like. In addition, each conversion module may be numbered to facilitate searching or replacement. For example, for a grouping of 2 rows by 6, the first row may be numbered 1, 3, 5, 7, 9, 11, and the second row may be numbered 2, 4, 6, 8, 10, 12.
In an alternative embodiment, a mounting module may be provided on the sample side; the installation module is provided with a sample side optical fiber, so that the sample side optical fiber corresponds to the sample hole conveniently.
In order to enable the sample side optical fibers to be neatly arranged, the installation module comprises M installation seats which are arranged side by side, and every N groups of sample side optical fibers are arranged on one installation seat. As shown in fig. 8, the sample well is exemplified by 96 wells, and the converted 8 sets of optical fibers a1, a2, a3, a4, a5, a6, a7, and a8 are mounted side by side on one mount, and 12 mounts are disposed side by side. Namely, each of the 12 excitation A, B … fiber bundles in the excitation cable and the 12 detection A ', B' … fiber bundles in the detection cable finally corresponds to one of the mounts.
It should be noted that the sample-side mounting seat shown in fig. 8 is not necessary, and 8 groups of converted sample-side optical fibers may be directly corresponding to one column of sample wells by other means, each column is 8 sample wells, so that each sample-side optical fiber or each bundle of sample-side optical fibers corresponds to one sample well, and the sample wells have 12 columns in total, as shown in fig. 9, the arrangement of the sample wells is 8 × 12.
In this embodiment, the sample side optical fibers are arranged in groups, so that each mounting seat or each row of sample holes corresponds to each group of sample side optical fibers, and the position of a certain sample side optical fiber is easy to find; in addition, when a certain sample side optical fiber is damaged, the optical fiber is easy to replace, and the structural complexity and the cost can be reduced.
In an optional embodiment, the excitation end and the detection end are respectively and independently encapsulated in different shells, specifically, the excitation light path and the emission light path are independently arranged, the excitation light path may include a light source, a convex lens, an excitation light filter wheel, an excitation end, an excitation optical fiber arranged in an excitation optical cable manner, and the like, and the emission light path may include a detector, a receiving optical filter, a detection end, a detection optical fiber arranged in a detection optical cable manner, and the like. When a certain optical path is in fault, the optical path can be independently replaced and maintained, and the maintenance cost is reduced.
In an alternative embodiment, the M excitation optical fiber bundles or the M detection optical fiber bundles have corresponding numbers, respectively, so that the positions of the excitation optical fibers or the detection optical fibers are easy to find; in addition, the optical fiber can be easily replaced when the excitation optical fiber or the detection optical fiber is damaged.
In all examples shown and described herein, any particular value should be construed as merely exemplary, and not as a limitation, and thus other examples of example embodiments may have different values.
The embodiment of the invention also provides a PCR detection method, which is applied to the PCR detection device and comprises the following steps:
(1) exciting light emitted by the exciting end is transmitted to the conversion module through the M exciting optical fiber bundles and then transmitted to the sample hole through the sample side optical fiber, and a sample to be detected is excited to obtain emitted light; wherein, each excitation optical fiber bundle is provided with N excitation optical fibers, M is more than or equal to 2, and N is more than or equal to 2;
specifically, excitation light emitted by the light source is focused by the convex lens, filtered by the excitation light filter wheel to generate excitation light with a specific wavelength, transmitted by the M excitation optical fiber bundles emitted by the excitation end, transmitted for a distance, enters the sample side optical fiber, and then converged by the lens group 9 to be emitted to the sample hole, so as to excite the sample to be detected to generate fluorescence signal emission light.
(2) Transmitting emitted light to a conversion module through a sample side optical fiber, and then transmitting the emitted light to a detector through M detection optical fiber bundles; wherein, each detection optical fiber bundle is provided with N detection optical fibers;
(3) the detector detects the intensity of the fluorescent signal of the emitted light.
Specifically, after entering the sample side optical fiber and being transmitted to the conversion module, the emitted light is projected to the detector 6 at the detection end through the M detection optical fiber bundles, and the fluorescence signal intensity of the emitted light is detected by the detector, and in addition, the emitted light can be filtered by the receiving optical filter 7 before entering the detector.
In this embodiment, the excitation end emits excitation light through M excitation fiber bundles, and each excitation fiber bundle is provided with N fibers, which are collectively M × N excitation fibers, and similarly, the detection end also includes M × N detection fibers, and after being converted by the conversion module, the M × N excitation fibers and the M × N detection fibers become M × N groups of sample side fibers capable of transmitting excitation light and receiving fluorescence.
And each N groups of sample side optical fibers correspond to one row of sample holes, so that the samples to be detected at the plurality of sample holes can be excited simultaneously. The column of sample wells may be N such that there is one sample well for each or each bundle of sample side fibres.
In an alternative embodiment, the M excitation fiber bundles may be transported over a distance at the excitation end in the form of an excitation optical cable and the M detection fiber bundles may be transported over a distance at the detection end in the form of a detection optical cable. Therefore, the excitation light emitted by the excitation end is transmitted to the conversion module through the excitation optical cable, and the emission light is transmitted to the conversion module through the sample side optical fiber and then transmitted to the detector through the detection optical cable.
Through the mode transmission of excitation optical cable and detection optical cable, can gather together M N excitation optic fibre and gather together M N detection optic fibre at the sense terminal, rather than come out from the port and just disperse, like this, can reduce the volume and the complexity of arranging of whole optic fibre greatly.
In the PCR detection method provided by the embodiment of the invention, excitation light is transmitted through M excitation optical fiber bundles at an excitation end, each excitation optical fiber bundle comprises N excitation optical fibers, emission light is transmitted through M detection optical fiber bundles at a detection end, and each detection optical fiber bundle comprises N detection optical fibers. High-throughput detection can be rapidly achieved through a large number of optical fibers. In particular, the effect is more remarkable when the number of sample holes is larger and the number of optical fibers is larger.
It is clear to those skilled in the art that, for convenience and brevity of description, specific implementation procedures of the foregoing method embodiments may be referred to in the foregoing device embodiments, and are not described herein again.
In the embodiments provided in the present application, it should be understood that the disclosed apparatus may be implemented in other ways. The above-described embodiments of the apparatus are merely illustrative, and for example, the division of the units is only one logical division, and there may be other divisions when actually implemented, and for example, a plurality of units or components may be combined or integrated into another system, or some features may be omitted, or not executed. In addition, the shown or discussed mutual coupling or direct coupling or communication connection may be an indirect coupling or communication connection of devices or units through some communication interfaces, and may be in an electrical, mechanical or other form.
In addition, in the description of the embodiments of the present invention, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.
In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc., indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, and are only for convenience of description and simplicity of description, but do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus, should not be construed as limiting the present invention.
Finally, it should be noted that: the above-mentioned embodiments are only specific embodiments of the present invention, which are used for illustrating the technical solutions of the present invention and not for limiting the same, and the protection scope of the present invention is not limited thereto, although the present invention is described in detail with reference to the foregoing embodiments, those skilled in the art should understand that: any person skilled in the art can modify or easily conceive the technical solutions described in the foregoing embodiments or equivalent substitutes for some technical features within the technical scope of the present disclosure; such modifications, changes or substitutions do not depart from the spirit and scope of the embodiments of the present invention, and they should be construed as being included therein. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.
Claims (10)
1. A PCR detection apparatus, comprising: the device comprises an excitation end, a detection end and a conversion module; the excitation end comprises M excitation optical fiber bundles, and each excitation optical fiber bundle is provided with N excitation optical fibers; the detection end comprises M detection optical fiber bundles, and each detection optical fiber bundle is provided with N detection optical fibers; wherein M is more than or equal to 2, and N is more than or equal to 2;
the conversion module converts one of the M excitation fiber bundles and one of the M detection fiber bundles into N groups of sample side fibers; each N sets of sample side fibers correspond to a column of sample wells.
2. The apparatus of claim 1 wherein M of said excitation fiber bundles are transported over a distance at said excitation end by means of excitation fiber optic cables and M of said detection fiber bundles are transported over a distance at said detection end by means of detection fiber optic cables.
3. The apparatus of claim 1, wherein said conversion modules comprise M, each said conversion module converting one said excitation fiber bundle and one said detection fiber bundle into N sets of sample side fibers, one set of said sample side fibers for each said sample well.
4. The apparatus of claim 3, further comprising a fiber mount on which M of the conversion modules are placed side-by-side.
5. The apparatus of claim 4, wherein M of the conversion modules are grouped into sets, each set being positioned side-by-side on the fiber mount.
6. The apparatus of claim 1, further comprising a sample side mounting module; the sample side optical fiber is installed on the installation module.
7. The apparatus of claim 6, wherein the mounting module comprises M side-by-side mounts, and every N sets of the sample-side optical fibers are arranged on one of the mounts.
8. The device of claim 1, wherein the excitation tip and the detection tip are each separately housed in separate housings.
9. The apparatus of claim 1, wherein each of the M excitation fiber bundles or the M detection fiber bundles has a corresponding number.
10. A PCR detection method applied to the PCR detection apparatus according to any one of claims 1 to 9, the method comprising:
exciting light emitted by the exciting end is transmitted to the conversion module through the M exciting optical fiber bundles and then transmitted to the sample hole through the sample side optical fiber, and a sample to be detected is excited to obtain emitted light; wherein each excitation optical fiber bundle is provided with N excitation optical fibers, M is more than or equal to 2, and N is more than or equal to 2;
the emitted light is transmitted to the conversion module through the sample side optical fiber and then transmitted to the detector through M detection optical fiber bundles; each detection optical fiber bundle is provided with N detection optical fibers;
the detector detects the intensity of the fluorescent signal of the emitted light.
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| CN202010118084.4A CN111307770A (en) | 2020-02-25 | 2020-02-25 | PCR detection device and method |
| PCT/CN2020/092278 WO2021169069A1 (en) | 2020-02-25 | 2020-05-26 | Pcr detection device and method |
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| CN113008854A (en) * | 2021-03-03 | 2021-06-22 | 艾普拜生物科技(苏州)有限公司 | Optical fiber array imaging detection device |
| CN113046230A (en) * | 2021-03-17 | 2021-06-29 | 杭州博日科技股份有限公司 | PCR instrument, detection method of PCR instrument and electronic terminal |
| CN117402729A (en) * | 2023-12-14 | 2024-01-16 | 鲲鹏基因（北京）科技有限责任公司 | PCR detection device |
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| CN116009142B (en) * | 2022-11-30 | 2023-07-25 | 南京春辉科技实业有限公司 | Medical PCR optical fiber bundle and multipath fluorescent quantitative PCR detector |
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Address after: Hangzhou City, Zhejiang province 310000 high tech Industrial Development Zone Waterfront Road 1192 Applicant after: Hangzhou bori Technology Co., Ltd Address before: 310000 no.1192 Bin'an Road, Binjiang District, Hangzhou City, Zhejiang Province Applicant before: HANGZHOU BIOER TECHNOLOGY Co.,Ltd. |
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Application publication date: 20200619 |