Machine for quantitative or qualitative PCR

Abstract

Machine for quantitative or qualitative PCR (1), characterized in that it comprises a viewing module (2) which includes at least one camera and is designed to monitor the position of the test tubes and the samples in the machine in order to signal to the user via display means if errors have been made in the positioning of the samples.

Description

[0001] The present invention relates to a machine for quantitative or qualitative PCR according to claim 1.

[0002] PCR (polymerase chain reaction) machines represent a milestone in molecular biology, as they enable the amplification and analysis of genetic material (DNA or RNA) in a biological sample. There are two main categories of PCR: qualitative and quantitative PCR.

[0003] Qualitative PCR is used to determine whether or not specific DNA or RNA sequences are present in a sample. This type of analysis is essential in fields such as medical diagnostics, where it is crucial to detect the presence of pathogens or genetic mutations. The qualitative PCR process involves several key steps. Initially, the DNA sample undergoes denaturation by heating, causing the separation of the two complementary strands. Specific probes or primers for the sequence of interest are then introduced and bind to the target DNA. Finally, a DNA polymerase synthesizes a new DNA chain complementary to the target, and the amplification cycle is completed. The results are analyzed using a dichotomous classification, indicating whether the target is present ("positive") or absent ("negative").

[0004] Quantitative PCR, also known as qPCR or real-time PCR, on the other hand, goes beyond the mere presence / absence of the target and allows the determination of its relative quantity in the initial sample. This method plays a crucial role in fields such as scientific research and disease diagnostics, where the quantity of genetic material can provide significant information about the biological state of the sample. qPCR differs from qualitative PCR in the use of fluorescent substances that bind to the DNA during amplification. The quantity of emitted fluorescence is proportional to the quantity of amplified DNA, allowing for real-time measurement of amplification. This technique offers remarkable precision in quantifying the genetic target present in the sample.

[0005] In short, PCR machines represent an essential component in molecular research and diagnostics, offering two complementary approaches: qualitative PCR, which determines the presence or absence of the target, and quantitative PCR, which provides a precise measurement of the quantity of the target in the sample. Both methods play a crucial role in advancing molecular biology and disease diagnosis.

[0006] PCR (Polymerase Chain Reaction) machines are complex devices whose construction is the result of sophisticated engineering that incorporates essential components to ensure the correct execution of the process.

[0007] The heart of a PCR machine is the thermocycler. This component is responsible for the precise and sequential control of temperatures during the denaturation, annealing, and elongation cycles.

[0008] Typically, the thermal cycler consists of an aluminum or copper block with heating elements and water or air cooling systems. These allow the temperatures required by the amplification protocol to be precisely reached and maintained.

[0009] The thermal cycler can incorporate a detection system that allows real-time monitoring of the accumulation of amplified DNA. This system utilizes fluorescence by incorporating a light source with one or more photocells. During the amplification phase, fluorescent markers are used to bind to the replicating DNA, generating a signal proportional to the quantity of genetic material produced.

[0010] An intuitive user interface is essential, allowing the operator to program and monitor the PCR run. This interface can include a display, control buttons, and dedicated data analysis software.

[0011] Finally, to ensure the efficiency and reliability of the processes, a PCR machine is equipped with safety and control systems to prevent errors or malfunctions. This can include temperature sensors, alarms, and emergency circuits.

[0012] Overall, a PCR machine represents a sophisticated example of the integration of electronic, thermal, and optical engineering, designed to deliver reliable and reproducible results in molecular analyses. Its precisely planned design and the integration of high-quality components are essential to ensuring the success of DNA or RNA amplification processes in the laboratory.

[0013] The throughput of a PCR machine can vary considerably depending on the specific model and device characteristics. Throughput refers to the quantity of samples that can be processed simultaneously or sequentially in a single run.

[0014] Some PCR machines are designed to analyze a single sample at a time, while others can process multiple samples in parallel. For example, some machines may have a 96- or 384-well thermal cycler, allowing for the simultaneous analysis of a corresponding number of samples. There are also more advanced platforms, such as those used in high-volume research and diagnostic applications, capable of processing even larger numbers of samples in a single run.

[0015] There are also high-performance PCR machines that can perform PCR cycles in parallel on multiple plates or thermal cyclers, thereby increasing throughput even further.

[0016] It should be noted that the throughput of a PCR machine can influence the workload that can be managed in a given period of time. For example, high-performance machines may be essential in diagnostic laboratories that need to analyze large sample volumes in a short time, while lower-throughput machines may be sufficient in a research setting with a lower workload.

[0017] Ultimately, the throughput of PCR machines is an important feature to consider when selecting the device that best suits a laboratory's specific needs.

[0018] Currently, the economic accessibility of PCR machines for small laboratories is often limited to relatively low-throughput machines. This means that these machines are capable of processing a limited number of samples simultaneously or sequentially. While these machines are essential for performing basic DNA or RNA amplification tasks, their low throughput can pose a significant challenge for laboratories with higher analysis volumes. The need to perform PCR cycles in series can lead to longer analysis times and less efficient management of the overall workload. This economic limitation can impact the productivity and responsiveness of such laboratories in the face of increased demand for molecular analysis, as was the case during the COVID-19 pandemic.Therefore, although low-throughput PCR machines represent a valuable access point for many laboratories, it is important to recognize that they can slow down the analysis process and that it would be appropriate to have medium-throughput but low-cost machines.

[0019] A particular problem with PCR machines is the risk of operator error in correctly positioning test tubes in the device. This type of error can occur for a variety of reasons, such as momentary distraction, inadequate lighting in the work area, or a lack of familiarity with the specific arrangement of samples in the machine. It is essential that the operator pays particular attention during this crucial phase of the procedure, as incorrect positioning of test tubes could compromise the accuracy and reliability of the amplification results. This eventuality underscores the importance of standardized training and procedures to ensure the correct and reliable execution of molecular analyses.

[0020] Currently, PCR machines do not have systems for automatically controlling the positioning of test tubes. This means that the operator bears sole responsibility for correctly positioning the test tubes within the device. There are no verification mechanisms or automatic detection systems that can prevent or correct any positioning errors. It is therefore essential that the operator exercise attention and care during this critical phase of the molecular analysis process to ensure the reliability and accuracy of the results obtained by amplifying the genetic material.

[0021] The aim of the present invention is to provide a machine for quantitative or qualitative PCR capable of providing a medium throughput and preventing errors in the positioning of the test tubes.

[0022] The particular solution of the present invention provides for a PCR machine provided with a vision module comprising at least one camera and designed to monitor the position of the test tubes and the samples in the machine in order to notify the user, via display means, in case of errors in the positioning of the samples.

[0023] The machine's components also make it possible to keep commercial costs low while ensuring a medium throughput.

[0024] Further advantages and characteristics of the invention will become clearer from the description of one of its possible embodiments, given purely as an example and not as limiting, with the aid of the accompanying drawings, in which: - Fig. Figure 1 is a front view of an open PCR machine according to the invention; - Fig. Figure 2 is an axonometric view of a PCR machine according to the invention without side walls for ease of illustration; - Fig. Figure 3 is an axonometric view of a closed PCR machine according to the invention; - Fig. Figure 4 is an exemplary schematic view of the vision module present in a PCR machine according to the invention;

[0025] In Fig. 1 shows a machine for quantitative or qualitative PCR 1, which in the illustrated embodiment contains inside a housing: - a test tube rack 6 suitable for holding the test tubes containing the sample to be analyzed, the controls and the standards; advantageously, the test tubes may be the same ones into which the sample to be analyzed has been taken, without the need for decanting or any preparatory operations; advantageously, a further separate rack may be provided for holding the controls and standards. - a mobile pipettor 7 capable of moving within the housing thanks to the action of its own moving means and suitable for extracting, with a tip, all or part of the samples to be analyzed from the test tubes in the receiving rack 6, in order to introduce them into an extraction module 8 equipped with cavities, suitable for carrying out the extraction and / or purification of the DNA / RNA by means of magnetic beads, magnetic bars being provided in the extraction module for moving the beads.

[0026] The sampling process is carried out thanks to the action of the pipettor's moving means, which allows the tip to aspirate the sample from the container (test tube). The pipettor is equipped with a suction system that creates a pressure differential, allowing the liquid to be drawn into the tip. Once the sample has been sampled, the pipettor can transfer it to other parts of machine 1, such as the extraction module or other phases of the molecular analysis process. This allows the sample to undergo further processing or analysis, such as DNA or RNA extraction and amplification by PCR. - a waste tray 9, into which the pipettor 7 can deposit its tip after extracting a sample. It is generally necessary to change the tips of the pipettors of a PCR machine 1 from one sample to another. This is essential to avoid cross-contamination between different samples, which could compromise the accuracy and validity of molecular analysis results. The use of disposable or washable tips, depending on the type of pipettor, is standard practice in molecular laboratories. Furthermore, changing tips also helps prevent any residues that could affect sample aspiration or dispensing during subsequent procedures. - a tip rack 10, suitable for holding the tips of the pipettor 7 and designed to remove one of them when necessary. The tip rack 10 can be designed to accommodate different types of tips, depending on the model of the machine 1 and the specific requirements of the laboratory; - a thermocycler 11 to complete the quantitative or qualitative PCR analysis from the samples contained in the extraction module 8 after extraction by the magnetic rod system. The main function of a thermocycler is to regulate and control the temperatures during the DNA or RNA amplification cycles. PCR requires heating and cooling cycles of the sample, as it is a process involving three phases: denaturation, annealing, and elongation. These phases require different temperatures to activate specific biochemical reactions. The thermocycler is designed to perform these heating and cooling cycles quickly and precisely. This process allows for the efficient amplification of the genetic material contained in the sample.Furthermore, in the case of quantitative PCR or “qPCR”, the thermal cycler 11 is capable of detecting the accumulation of amplified DNA in real time and provides quantitative data on the quantity of genetic target present in the sample. - an electronic control unit suitable for receiving the data in the thermocycler and coordinating the operability of the various elements of the machine 1.

[0027] The PCR machine 1 according to the invention is capable of performing analyses using directly the sample in the tubes used for collection from the patient.

[0028] To ensure accurate positioning of the test tubes and avoid potential human errors, the PCR machine 1 is equipped with an advanced vision module 2 that includes at least one camera. This component plays a crucial role in the system's operability, continuously and constantly monitoring the correct arrangement of the test tubes in the machine 1. The camera records images of the test tubes in real time, allowing the system to provide immediate feedback to the operator if potential positioning errors are detected.

[0029] The vision module 2 is preferably configured to execute algorithms capable of precisely analyzing and distinguishing the shapes of the test tubes present in the machine 1. These algorithms operate by processing the images captured by the camera and are capable of determining whether the test tubes are positioned correctly or not. For this purpose, the vision module 2 advantageously uses a predefined set of correct positions contained in its own database. During operation, the captured images are constantly compared with these sets of ideal positions. If a discrepancy is detected, the system is capable of indicating it to the operator, allowing timely correction of the test tubes' position.

[0030] This technological approach is important for guaranteeing the accuracy and reliability of the molecular analyses performed by the PCR machine 1, as it minimizes the possibility of malfunctions or false results due to improper positioning of the test tubes.

[0031] Feedback is provided via a suite of specially designed indicators, including acoustic, haptic, and optical systems. In the event of improper positioning of the test tubes, these devices are activated, alerting the operator with acoustic and visual signals and tactile feedback. This synergy between the vision module 2 and the indicators ensures an immediate and efficient response if a positioning error is detected. Thanks to this control and notification system, greater precision and reliability are ensured in the molecular analysis process performed by the PCR machine 1.

[0032] The PCR machine 1 is further equipped with a display 3, which can be implemented with touchscreen technology. This instrument provides the user with an intuitive and interactive interface that facilitates programming and monitoring procedures. Through this display 3, the operator can navigate between the various configuration options, adjust the execution parameters, and monitor the analysis status in real time. Its intuitive interface makes the operation of the PCR machine 1 accessible even to less experienced users, improving the efficiency and ease of use of the device. This instrument, together with the other control and monitoring components, helps ensure that the molecular analysis is carried out precisely and reliably and can be used by the vision module 2 as a display to provide the user with visual feedback.The display 3 allows the sequence of procedures to be performed to be displayed, verifying the correct positioning of the various elements and detecting and signaling any positioning errors. Advantageously, the PCR machine 1 includes a reader for identification codes 4, such as barcodes, which plays a key role in ensuring precise sample traceability during the various analysis phases. This device is designed to read the identification codes applied to the test tubes containing the samples, allowing precise identification and registration of each sample in the system. Thanks to this technology, it is possible to monitor in detail the movements of the test tubes in the machine 1, guaranteeing reliable and accurate sample management throughout the entire molecular analysis process.This functionality translates into a significant reduction in the risk of errors and complete traceability, which are essential elements for the quality and reliability of the molecular analyses performed by PCR Machine 1.

[0033] The PCR machine 1 advantageously also includes a temperature monitoring and control system, enabled by the use of dedicated sensors. These sensors are strategically positioned to accurately detect ambient temperature changes. The information acquired by the sensors is then transmitted to the electronic control unit. This control unit is specifically designed to process the data from the sensors and activate the heating / cooling means to precisely regulate the temperature conditions in the machine 1, maintaining them consistently within the desired ranges to perform accurate and reliable PCR.

[0034] The PCR machine 1 is also optionally equipped with a HEPA (High Efficiency Particulate Air) filter 5, which plays a crucial role in maintaining a clean working environment. This filter is designed to capture microscopic particles, including potential contaminants such as cells, bacteria, or viruses present in the ambient air. Its high efficiency in capturing such particles guarantees a contaminant-free environment, which is essential for the accuracy and reliability of the molecular analyses performed in the machine 1. Thanks to the HEPA filter 5, the risk of cross-contamination between samples is minimized, ensuring precise and reliable results at every stage of the amplification process.

[0035] Advantageously, at least one UV lamp 15 may also be included, always to guarantee environmental safety.

[0036] From the above description, it is clear that the PCR machine 1 according to the invention is capable of solving the known problems of the prior art by simplifying the analysis process and reducing the risk of errors. Furthermore, the special components used enable good throughput at a low cost, thus increasing productivity even for small / medium-sized analysis laboratories.

[0037] Although the inventive concept has been described with reference to exemplary embodiments, it will be apparent to those skilled in the art that various changes and modifications may be made without departing from the scope of the inventive concept. Therefore, it should be understood that the above embodiments are not limiting but illustrative. The invention is defined by the following claims.

Claims

[1] Machine for quantitative or qualitative PCR (1), characterized by in that it comprises a vision module (2) which comprises at least one camera and is designed to monitor the position of the test tubes and the samples in the machine in order to signal to the user via display means if errors have been made in the positioning of the samples. [2] Machine for quantitative or qualitative PCR (1) according to claim 1, characterized by that it contains in one housing: - a test tube rack (6) suitable for holding the test tubes containing the samples to be analyzed, the controls and the standards; - a mobile pipettor (7) capable of moving within the housing thanks to the action of its own moving means and suitable for extracting, with a tip, all or part of the samples to be analyzed from the test tubes in the receiving rack (6) in order to introduce them into an extraction module (8) equipped with cavities, suitable for carrying out the extraction and / or purification of the DNA / RNA by means of magnetic beads, magnetic bars being provided in the extraction module for moving the beads. - a waste tray (9) into which the pipettor (7) can eject its tip after extracting a sample. - a tip holder rack (10) suitable for holding the tips of the pipettor (7) and designed to remove one of them when necessary. - a thermocycler (11) to complete the quantitative or qualitative PCR analysis starting from the samples contained in the extraction module (8) after extraction by the magnetic rod system. - an electronic control unit suitable for receiving the data in the thermal cycler and coordinating the operability of the various elements of the machine. [3] Machine for quantitative or qualitative PCR (1) according to any one of the preceding claims, characterized by that it comprises a display (3) to enable the user to interact with the machine, to show the user the results of the analysis and to signal any errors in the positioning of the samples detected by the vision module (2). [4] Machine for quantitative or qualitative PCR (1) according to any one of the preceding claims, characterized bythat it comprises an identification code reader (4) suitable for reading the identification codes present on the test tubes. [5] Machine for quantitative or qualitative PCR (1) according to any one of the preceding claims, characterized by that it includes a HEPA filter (5). [6] Machine for quantitative or qualitative PCR (1) according to any one of the preceding claims, characterized by that it comprises at least one UV lamp (15). [7] Machine for quantitative or qualitative PCR (1) according to any one of the preceding claims, characterized by that it comprises a temperature monitoring and control system comprising at least one temperature sensor and heating / cooling means. [8] Machine for quantitative or qualitative PCR (1) according to any one of the preceding claims, characterized bythat it provides a separate rack for the controls and standards.

Images