Publish Time: 2022-01-24 Origin: Site
The characteristics of the management system are that it should have a clear purpose, standardized management, effective restraint, efficient mechanism, and self-development as a whole.
A well-established SOP document shall be established to specify the method for accomplishing various quality activities, and each SOP shall describe one or a group of interrelated activities.
Each SOP document shall describe the documents, materials, personnel, records required for the input, transformation and output of the quality link and their interface with the relevant activities.
Clarify the requirements of various factors in the conversion process of each link, that is, who will do it, what will be done, what procedures will be done, what requirements will be met, how to control, what records and reports will be formed, and the corresponding approval procedures. Corrective actions that specify exceptions or special circumstances that require attention in quality activities. SOPs should be concise, clear and understandable and should be mastered and strictly followed by staff.
To complete a set of PCR experiments, it usually requires four experimental processes: reagent preparation, sample processing, nucleic acid amplification and product analysis.
The experimental rooms for the four experimental processes should be arranged adjacent to each other to form an independent PCR experimental area. The standard PCR experimental area includes: reagent preparation area, sample preparation area, amplification area, product analysis area, supporting buffer area and public corridor in each area.
1. Reagent preparation area
The main operations in this experimental area are the preparation of storage reagents, the sub-packaging of reagents and the preparation of the main reaction mixture. Reagents and materials used for specimen preparation should be shipped directly to this area and must not pass through other areas. The reagent raw materials must be stored in this area, and the required storage reagents must be prepared in this area. The reagent preparation area is equipped with a 2~8°C refrigerator and a -80°C refrigerator, which need to be included when calculating the cooling load. The area of the room should be controlled within 15m2~20m2. The pressure gradient requirements in this area are: a relatively positive pressure state to prevent the entry of outside air containing nucleic acid aerosols and cause pollution.
2. Specimen preparation area
The main operations in this area are the preservation of clinical specimens, nucleic acid (RNA, DNA) extraction, storage, and cDNA synthesis when adding to amplification reaction tubes and measuring RNA.
The specimen preparation area is equipped with a 2~8°C refrigerator and a -20°C refrigerator, which needs to be included when calculating the cooling load. In the specimen preparation area, a biological safety cabinet is also required for the extraction of nucleic acid. In order to avoid repeated circulation of the extracted nucleic acid in the cabinet, resulting in cross-contamination between specimens and false positive results, the biological safety cabinet equipped in this area must be of type B2. The vertical airflow in the working area of the biological safety cabinet is all from the laboratory, and the exhaust air is filtered by the high-efficiency filter and directly discharged to the outside, and is not allowed to return to the safety cabinet and laboratory. According to experience, if the laboratory is equipped with biological safety cabinets, the area of the laboratory will increase by 10m2 for each set; the area of the specimen preparation area should be between 25m2 and 30m2. The pressure gradient of this area is required to be positive pressure relative to the adjacent area, so as to avoid aerosol pollution from the adjacent area into this area.
3. Amplification chamber
The main operations performed in this region are DNA or cDNA amplification. In addition, the addition of prepared DNA templates and synthesized cDNA (from the sample preparation area) and the preparation of the main reaction mixture (from the reagent storage and preparation area) into a reaction mixture, etc., can also be performed in this area.
The amplification room is mainly equipped with a PCR instrument, the core instrument of the PCR laboratory. In the process of laboratory construction, it is necessary to equip the PCR instrument with a special UPS power supply to ensure its normal operation. The area of this area is controlled at 15m2~20m2.
The pressure gradient in this area is required to be negative pressure relative to the adjacent area to avoid aerosol leakage from this area. To avoid pollution caused by aerosols, unnecessary movement in the area should be minimized. Individual operations such as sample addition should be carried out in a clean bench. It is recommended to use 5~10Pa differential pressure, which is easier to achieve in control.
4. Product analysis area
The main operation in this area is the determination of amplified fragments. This area can be omitted if the automatic closed analytical instrument is used for detection.
Equipped with a fume hood indoors to ensure a relative negative pressure in the room, and the air flows from the outdoors to the indoors. The area of this area is controlled at 15m⊃2;~20m⊃2;. This area is the main source of contamination of amplification products, so the requirement for the pressure gradient in this area is: a negative pressure relative to the adjacent area, so as to avoid the diffusion of amplification products from this area to other areas. It is recommended to use 5~10Pa differential pressure, which is easier to achieve in control.