CN-121972072-A - Automatic liquid mixer for detecting virus titer based on plaque method

CN121972072ACN 121972072 ACN121972072 ACN 121972072ACN-121972072-A

Abstract

The invention discloses an automatic liquid mixer for detecting virus titer based on a plaque method, which comprises a workbench, a first constant temperature module, a second constant temperature module, a host cell liquid fixing module, a virus test sample fixing module and an execution module, wherein the first constant temperature module, the second constant temperature module, the host cell liquid fixing module, the virus test sample fixing module and the execution module are integrated on the workbench, the first constant temperature module and the virus test sample fixing module are provided with space intervals, the execution module is provided with a first moving path, a second moving path and a third moving path, the virus test sample fixing module and the first moving path are arranged in a staggered mode, the host cell liquid fixing module and the second moving path are arranged in a staggered mode, the positions of the modules are reasonably arranged, and the execution module is utilized to automatically execute liquid transferring, liquid dividing and mixing operations, so that the mixed liquid proportion is accurate, the detection result is accurate, and the operation efficiency is improved.

Inventors

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Assignees

杭州纽创生物检测有限公司

Dates

Publication Date: 20260505
Application Date: 20251231

Claims (10)

1. An automatic liquid mixer for detecting virus titer based on plaque method is characterized by comprising a workbench, a first constant temperature module, a second constant temperature module, a host cell liquid fixing module, a virus test sample fixing module and an execution module, wherein the first constant temperature module, the second constant temperature module, the host cell liquid fixing module, the virus test sample fixing module and the execution module are integrated on the workbench; The first constant temperature module is used for preserving heat of mixed liquid in the mixing tube; the second constant temperature module is used for preserving the temperature of the liquid agar in the agar bottle; the host cell liquid fixing module is used for placing a host cell liquid bottle; the virus test sample fixing module is used for placing a container containing a virus test sample; the first constant temperature module and the virus test sample fixing module are provided with a space interval; The execution module is provided with an execution end which can form detachable sealing connection with the suction head and controls the suction head to automatically suck, carry and release host cell liquid or virus test sample or liquid agar; the execution module is provided with a first moving path, a second moving path and a third moving path; On the first moving path, the execution module is sucked by the host cell liquid fixing module through a suction head and carries host cell liquid to move to the first constant temperature module, and the host cell liquid is injected into the mixing pipe; on the second moving path, the execution module sucks and carries the virus test sample to the first constant temperature module through a suction head by the virus test sample fixing module, and injects the virus test sample into the mixing tube; The virus test sample fixing module and the first moving path are arranged in a staggered manner, and the host cell liquid fixing module and the second moving path are arranged in a staggered manner; And on the third moving path, the executing module sucks and carries the liquid agar to the first constant temperature module through the suction head by the second constant temperature module, and injects the liquid agar into the mixing tube.
2. The automated mixer of claim 1, wherein the first path has a minimum length L1, the second path has a minimum length L2, L1< L2 is satisfied, and/or 200mm < L2 < 500mm.
3. The automated pipetting machine of claim 2 wherein the host cell fluid holding module and the virus test sample holding module are each located on either side of the first constant temperature module and/or 30 mm≤L1≤80 mm.
4. An automated mixer according to any one of claims 1-3, further comprising a tip retrieval module for collecting used tips, the execution module further having a fourth path of movement on which the execution module is moved from the first thermostatic module to the tip retrieval module and the used tips are thrown into the tip retrieval module; the second constant temperature module, the host cell liquid fixing module and the virus test sample fixing module are arranged in a staggered mode with the fourth moving path.
5. The automated fluidic mixer of claim 4, wherein the second thermostatic module is located between the first thermostatic module and the virus-test-sample fixing module, or wherein the tip recovery module is located between the first thermostatic module and the virus-test-sample fixing module.
6. The automated fluidic mixer of claim 1, wherein the second thermostatic module is located between the first thermostatic module and the virus test sample fixing module, and the third movement path has a minimum length L3, wherein 30mm ∈l3 ∈80mm.
7. The automated liquid mixing machine of claim 4, further comprising a tip module comprising a tip rack and a plurality of unused tips inserted into the tip rack; The execution module is provided with a fifth moving path, and the execution module moves to the suction head module from the suction head recovery module to acquire unused suction heads on the fifth moving path; the suction head module, the first moving path, the second moving path and the fourth moving path are arranged in a staggered mode.
8. The automated mixer of claim 7, wherein the tip module and the second thermostatic module are located on either side of the first thermostatic module.
9. The automated fluidic mixer of claim 7, wherein the tip module and the tip recovery module are on the same side of the first thermostatic module and/or the tip module and the host cell fluid immobilization module are on the same side of the first thermostatic module.
10. The automated pipetting machine of claim 1 wherein a cabinet is positioned above the table and wherein the first constant temperature module, the second constant temperature module, the host cell fluid holding module, the virus test sample holding module, and the actuator module are positioned within a space enclosed by the cabinet and/or, The execution module comprises a liquid transfer device, a mechanical arm, a linear actuator and a controller, wherein the liquid transfer device is arranged at the execution end of the mechanical arm, the mechanical arm is movably arranged on the linear actuator, the controller is used for controlling the execution module to execute liquid taking operations and corresponding moving paths, and the linear actuator is fixed on the workbench.

Description

Automatic liquid mixer for detecting virus titer based on plaque method Technical Field The invention relates to the technical field of virus detection, in particular to an automatic liquid mixer for detecting virus titer based on a plaque method. Background In the processing technology of biological agents, virus removal has become an indispensable step to meet the virus safety requirements. In the production process of the biological agent, virus inactivation and/or virus removal procedures are required, and a common virus removal method is to remove viruses through interception of a filter membrane, and the virus removal capacity of the filter membrane directly determines the virus safety of the biological agent. The membrane separation technology uses a filter membrane as a separation medium, when a certain driving force (such as pressure difference, concentration difference and the like) exists on two sides of the filter membrane, raw material components selectively permeate the filter membrane, and viruses are physically trapped due to the fact that the size of the raw material components is larger than the aperture of the filter membrane. Plaque method based on virus infectivity detection of virus titer is classical virus titer detection technique, the basic principle is that after host cells are cultured to a monolayer, properly diluted virus is inoculated and cultured, virus is adsorbed and invaded into host cells, and enzyme in the host cells is utilized to synthesize self-components, so that the host cells are cracked and dead, and plaque is formed, and one plaque represents one living virus particle. For example, the American society PDA (Parenteral Drug Association) refers to the biological agent's Virus Filtration test method and the manual file "Virus Filtration," TECHNICAL REPORT No.41 (recycled 2008), abbreviated as TR41, issued for Virus removal verification. The TR41 document states that the entrapped virus is phage PR772 or PP7 (PR 772 is a large virus, PP7 is a small virus, optionally) and the material stream is immunoglobulin IVIG (or bovine serum albumin BSA). Titer detection of both PR772 phage and PP7 phage was achieved using the plaque method described previously. More specifically, the minivirus-entrapment filter-test protocol, which uses PP7 phage as a nominally challenge virus model, is well documented in appendix III of TR41, with human immunoglobulin (IVIG) for protein transmittance assessment. To calculate the viral retention, the titers of PP7 phage in the challenge and filtrate, respectively, were measured. The method for detecting the titer of PP7 phage is well described in TR41, and the samples are counted by culturing using a double-layer plate counting method, wherein the specific formulation of the detection system is 1mL of a filtrate sample containing PP7 phage and 2mL of host cells (Pseudomonas aeruginosa) at a specific concentration, and 9mL of warm agar, or about 0.1mL of a filtrate sample containing PP7 phage, 1mL of host bacteria and 4.5mL of agar solution. At present, the operations of liquid preparation and liquid mixing of host cell liquid, bacteriophage and soft agar are completely carried out manually, the detection quantity is limited by manual work, the efficiency is extremely low, and the detection requirement cannot be met. In addition, for the method for detecting virus titer in the ultralow virus titer system as disclosed in the prior patent CN115181816B, since the titer of PP7 in the filtrate is lower than the detection limit, in order to ensure the accuracy of the detection result, a method is adopted in which a filtrate with a sufficiently large detection volume must be accumulated for a certain volume of filtrate, i.e. the detected filtrate volume must be sufficiently large. For example, the volume of the filtrate is 100ml, and if only 1ml is detected, the probability of detecting 0 pp7 is relatively high, and the detection result has no statistical significance, so that at least 30ml of filtrate is required to be detected, 30 times of mixing are required, and the workload is extremely high. Based on this, it is necessary to design an automatic liquid mixer for virus titer detection based on plaque method to improve detection efficiency and provide high-precision detection results. Disclosure of Invention Aiming at the defects existing in the prior art, the invention aims to provide an automatic liquid mixer for detecting virus titer based on a plaque method, which solves the problem of lower efficiency of the existing plaque method-based virus titer detection. In order to achieve the above purpose, the present invention adopts the following technical scheme: an automatic liquid mixer for detecting virus titer based on plaque method comprises a workbench, a first constant temperature module, a second constant temperature module, a host cell liquid fixing module, a virus test sample fixing module and an execution module, wherein the first constant