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CN-122016614-A - Flow type fluorescence analyzer and flow type fluorescence analysis method

CN122016614ACN 122016614 ACN122016614 ACN 122016614ACN-122016614-A

Abstract

The application relates to the technical field of biomedical detection, in particular to a flow type fluorescence analyzer and a flow type fluorescence analysis method. The flow type fluorescence analyzer comprises a pretreatment module, a magnetic bead enrichment and resuspension module and a flow type detection module, wherein the pretreatment module comprises a plurality of independent reaction container stations for respectively bearing independent samples to be detected and corresponding single-index fluorescent coding magnetic beads and performing independent immune reaction so as to obtain a reaction product suspension containing the fluorescent coding magnetic beads carrying the objects to be detected. The magnetic bead enrichment and resuspension module comprises at least one enrichment container and a magnetic field generating device, wherein the same enrichment container is used for receiving a plurality of reaction product suspensions from the pretreatment module, the magnetic field generating device is used for generating or removing a magnetic field at the enrichment container so as to enrich fluorescent coding magnetic beads carrying an object to be detected in the plurality of reaction product suspensions and obtain a magnetic bead suspension to be detected, and the flow detection module is used for detecting fluorescent signals of the fluorescent coding magnetic beads.

Inventors

  • ZHANG HONGXIANG
  • KUANG YUJI
  • LI WEIGONG

Assignees

  • 深圳唯公生物科技有限公司

Dates

Publication Date
20260512
Application Date
20260209

Claims (10)

  1. 1. A flow-type fluorescence analyzer, comprising: The pretreatment module (11) comprises a plurality of independent reaction container stations (111) which are used for respectively bearing independent samples to be tested and corresponding single-index fluorescent coded magnetic beads and carrying out independent immune reaction so as to obtain a plurality of reaction product suspensions containing fluorescent coded magnetic beads carrying the objects to be tested; A magnetic bead enrichment and resuspension module (12) comprising at least one enrichment vessel (121) and a magnetic field generating device (122), wherein the same enrichment vessel (121) is used for receiving a plurality of parts of the reaction product suspension from the pretreatment module (11), the magnetic field generating device (122) is arranged outside the enrichment vessel (121) and is used for generating or removing a magnetic field at the enrichment vessel (121) so as to enrich the fluorescent coding magnetic beads carrying the to-be-detected objects in the plurality of parts of the reaction product suspension, and the same enrichment vessel (121) is also used for accommodating to-be-detected magnetic bead suspensions containing different fluorescent coding magnetic beads carrying the to-be-detected objects; The flow type detection module (13) is in fluid communication with the magnetic bead enrichment and resuspension module (12), the flow type detection module (13) comprises a detection flow channel (131), a laser (132) and a photoelectric detector (133), the detection flow channel (131) is used for enabling a plurality of different fluorescent coding magnetic beads carrying a to-be-detected object in the to-be-detected magnetic bead suspension to sequentially pass through a laser irradiation area, the laser (132) is used for irradiating the fluorescent coding magnetic beads carrying the to-be-detected object passing through the laser irradiation area so as to excite fluorescent signals, and the photoelectric detector (133) is used for collecting different fluorescent signals emitted by the fluorescent coding magnetic beads carrying the to-be-detected object and at least identifying classified fluorescent signals for identifying magnetic bead codes and reporting fluorescent signals for reflecting the concentration of the to-be-detected object, wherein the fluorescent signals are included.
  2. 2. The flow fluorescence analyzer of claim 1, wherein the bead enrichment and resuspension module (12) further comprises a liquid collection assembly (123), the liquid collection assembly (123) comprising a liquid collection conduit (1231), a switching valve (1232), a top tube set (1233) and a drain tube set (1234), the liquid collection conduit (1231) comprising a suction end (12311) and a communication end (12312), the suction end (12311) extending into the enrichment vessel (121), the communication end (12312) optionally communicating with the top tube set (1233) or the drain tube set (1234) via the switching valve (1232) for introducing a different suspension of the reaction products into the enrichment vessel (121) or withdrawing liquid from the enrichment vessel (121).
  3. 3. The flow fluorescence analyzer according to claim 2, wherein the enriching container (121) is made of titanium alloy, the enriching container (121) comprises a bottom wall (1211) and a side wall (1212), the bottom wall (1211) and the side wall (1212) are matched to form an enriching cavity (1213), the magnetic field generating device (122) comprises a magnetic component (1221) and a driving component (1222), the magnetic component (1221) is in transmission connection with the driving component (1222), the driving component (1222) is used for driving the magnetic component (1221) to approach or separate from the side wall (1212), the magnetic component (1221) is used for attracting the fluorescent coded magnetic beads carrying the to-be-detected object to the inner surface of the side wall (1212) through magnetic force, and the sucking end (12311) extends into the enriching cavity (1213), and an opening of the sucking end (12311) faces the bottom wall (1211).
  4. 4. The flow fluorescence analyzer of claim 3, wherein the bead enrichment and resuspension module (12) further comprises an ultrasonic generation assembly (124), wherein the ultrasonic generation assembly (124) is connected with the bottom wall (1211), and the ultrasonic generation assembly (124) is used for generating ultrasonic waves to drive the different fluorescent-encoded magnetic beads carrying the object to be measured in the magnetic bead suspension to be measured to be uniformly mixed and resuspended.
  5. 5. The flow fluorescence analyzer of claim 3, wherein a side of the bottom wall (1211) remote from the magnetic field generating device (122) is configured to be disposed obliquely downward in a gravitational direction and cooperates with the side wall (1212) to form a suction zone (1214), the suction end (12311) extending into the suction zone (1214).
  6. 6. The flow fluorescence analyzer of claim 3, wherein the driving member (1222) comprises a driving member (1223), a sliding seat (1224), a sliding rail (1225) and a fixing plate (1226), the driving member (1223) and the enrichment vessel (121) are respectively connected to two opposite sides of the fixing plate (1226), the sliding rail (1225) is fixedly connected with the fixing plate (1226), the sliding seat (1224) is slidingly connected with the sliding rail (1225), the magnetic member (1221) is fixedly connected with the sliding seat (1224), the driving member (1223) is in driving connection with the sliding seat (1224) and is used for driving the magnetic member (1221) to approach or separate from the side wall (1212), and the enrichment vessel (121) further comprises a cantilever (1217), one end of the cantilever (1217) is connected with the side wall (1212) and the other end of the cantilever (1217) is connected with the fixing plate (1226).
  7. 7. A flow fluorescence analysis method, characterized in that the flow fluorescence analyzer according to any one of claims 1 to 6 is used, the flow fluorescence analysis method comprising the steps of: S1, in a plurality of independent reaction container stations (111) of the pretreatment module (11), respectively carrying out independent immunoreaction on a sample to be tested and corresponding single-index fluorescent coding magnetic beads so as to obtain a plurality of reaction product suspensions containing fluorescent coding magnetic beads carrying an object to be tested; S2, a step of enriching and re-suspending, namely introducing a plurality of reaction product suspensions into the same enriching container (121) of the magnetic bead enriching and re-suspending module (12), generating a magnetic field through the magnetic field generating device (122) to enrich fluorescent coding magnetic beads in the plurality of reaction product suspensions, and re-suspending the fluorescent coding magnetic beads to form a magnetic bead suspension to be detected containing different fluorescent coding magnetic beads in the same enriching container (121); s3, conveying the magnetic bead suspension to be detected to the flow detection module (13), enabling a plurality of different fluorescent coding magnetic beads in the magnetic bead suspension to be detected to sequentially pass through a laser irradiation area, exciting by the laser (132) and collecting fluorescent signals by the photoelectric detector (133), and identifying classified fluorescent signals for identifying magnetic bead codes and report fluorescent signals for reflecting the concentration of an object to be detected, which are included in the fluorescent signals.
  8. 8. The flow fluorometric assay of claim 7, wherein the enriching and resuspension step further comprises: introducing a plurality of reaction product suspensions into the same enrichment vessel (121) one or more times by utilizing a liquid collecting component (123), so that the magnetic field generating device (122) generates a magnetic field to enrich the fluorescent coding magnetic beads carrying the objects to be detected; After the magnetic field generating device (122) generates a magnetic field and constrains the positions of the fluorescent coding magnetic beads carrying the objects to be detected, the liquid in the enrichment container (121) is pumped out by the liquid collecting component (123); adding a resuspension liquid or retaining at least part of the liquid in the reaction product suspension added last time to the enrichment container (121), removing the magnetic field, and mixing and resuspension the enriched fluorescent coding magnetic beads to form the magnetic bead suspension to be tested.
  9. 9. The method according to claim 8, wherein in the enrichment and resuspension steps, the fluorescent-encoded magnetic beads carrying the analyte are magnetically attracted to the inner surface of the sidewall (1212) of the enrichment vessel (121), and an ultrasonic wave is generated by an ultrasonic generating assembly (124) to drive the fluorescent-encoded magnetic beads to uniformly suspend to achieve mixed resuspension.
  10. 10. The flow fluorometric assay of claim 7, wherein in the enriching and resuspension step, the multiple reaction product suspensions are introduced by: Sequentially introducing a plurality of parts of the reaction product suspension into the enrichment vessel (121), performing a magnetic field enrichment and a liquid withdrawal operation once after each of the introduced parts of the reaction product suspension, or After all of the reaction product suspensions are pooled into the enrichment vessel (121), one-time magnetic field enrichment and liquid withdrawal operations are performed.

Description

Flow type fluorescence analyzer and flow type fluorescence analysis method Technical Field The application relates to the technical field of biomedical detection, in particular to a flow type fluorescence analyzer and a flow type fluorescence analysis method. Background In clinical diagnostics, it is often necessary to detect multiple biomarkers, such as a set of cytokines, autoimmune antibodies or allergens, in a blood or body fluid sample simultaneously in order to fully assess the physiological or pathological state of a patient. The multi-index combined detection has important clinical value for early screening, diagnosis, disease condition monitoring and prognosis judgment of diseases. At present, two main flow technical paths mainly exist for realizing multi-index joint detection. The first is a single index detection technology represented by chemiluminescence immunoassay, and has the advantages that each detection item uses an independent kit and a reaction flow, the reaction condition is specific, the result is accurate and reliable, and single charging can be flexibly carried out on the detection item according to medical advice. However, when a sample needs to detect multiple indexes, repeated detection needs to be performed on the sample, so that the whole detection flux is low, the sample turnaround time (TAT) is obviously prolonged, and the requirement of a modern laboratory on high efficiency is difficult to meet. The other is a liquid phase chip (or flow type multiple immune analysis) technology based on a flow type fluorescence technology, and the technology can obtain results of tens of indexes simultaneously in one reaction and one detection of one sample by using microspheres with different fluorescence codes as carriers, and has the remarkable advantages of high flux, low cost and sample saving. However, existing conventional flow multiplex detection methods typically pre-mix all items of capture microspheres into a fixed detection kit. This mode cannot meet the flexibility requirements of detecting and charging for clinical split projects on demand, and if various possible combination packages are developed to meet the flexibility, huge research and development, registration and inventory management burdens are brought to reagent manufacturers. More importantly, a plurality of antibodies and antigens aiming at different analytes are incubated in the same reaction system, so that nonspecific interference such as cross reaction, steric hindrance and the like is extremely easy to cause, the sensitivity and accuracy of detection can be influenced, and meanwhile, each single detection is difficult to strictly follow independent and regulated optimized reaction conditions. How to solve the above problems is considered by those skilled in the art. Disclosure of Invention In order to solve the problems in the prior art, the application provides a flow type fluorescence analyzer and a flow type fluorescence analysis method. The embodiment of the application provides a flow type fluorescence analyzer, which comprises a pretreatment module, a magnetic bead enrichment and resuspension module and a flow type detection module. The pretreatment module comprises a plurality of independent reaction container stations which are used for respectively carrying independent samples to be tested and corresponding single-index fluorescent coding magnetic beads and carrying out independent immune reaction so as to obtain a plurality of reaction product suspensions containing the fluorescent coding magnetic beads carrying the objects to be tested. The magnetic bead enrichment and resuspension module comprises at least one enrichment container and a magnetic field generating device, wherein the same enrichment container is used for receiving multiple reaction product suspensions from the pretreatment module, the magnetic field generating device is arranged outside the enrichment container and used for generating or removing a magnetic field at the enrichment container so as to enrich fluorescent coding magnetic beads carrying an object to be detected in the multiple reaction product suspensions, and the same enrichment container is also used for accommodating the magnetic bead suspensions to be detected containing different fluorescent coding magnetic beads carrying the object to be detected. The flow type detection module is in fluid communication with the magnetic bead enrichment and resuspension module, and comprises a detection flow passage, a laser and a photoelectric detector, wherein the detection flow passage is used for enabling a plurality of different fluorescent coding magnetic beads carrying an object to be detected in a magnetic bead suspension to sequentially pass through a laser irradiation area, the laser is used for irradiating the fluorescent coding magnetic beads carrying the object to be detected passing through the laser irradiation area so as to excite fluorescent signals, and the photoele