Clinical PCR laboratory design

- Aug 06, 2020-

n recent years, new outbreaks of infectious diseases have occurred in the world, especially the new crown pneumonia, which has strong transmission capacity, fast transmission speed, wide range of infections, and high levels of infection. It has become an important area in global public health and is in the fight against new crown pneumonia. In the epidemic, large-scale nucleic acid testing has determined that the infected population is the top priority. The state has vigorously built clinical PCR laboratories in hospitals and epidemic prevention stations in various cities and counties for the new crown pneumonia. Next, I will introduce what is a clinical PCR laboratory.


       Clinical gene amplification testing laboratory refers to a laboratory that detects specific DNA or RNA through amplification for disease diagnosis, treatment, treatment, and prognosis determination. The clinical PCR laboratory must pass the inter-laboratory quality evaluation organized by the Ministry of Health Clinical Inspection Center or a designated institution before it can operate. Temporary laboratory personnel must pass the technical training of institutions designated by the provincial health administrative department before they can engage in clinical gene amplification testing.

PCR laboratory settings ●●




       The PCR laboratory is divided into four areas in principle: reagent preparation area, specimen preparation area, amplification area, and product analysis area. If using an automatic amplification analyzer, the latter two areas can be combined. If using a fully automatic analysis system that integrates nucleic acid extraction, amplification and product analysis, the latter three areas can be combined.


       With the popularity of real-time fluorescent quantitative PCR machines, most PCR laboratories are now divided into three areas: reagent preparation area (area one), specimen preparation area (area two), and amplification area (area three).


The design of the clinical PCR laboratory is tailored to local conditions, but it is necessary to ensure that each area has an independent buffer zone, each area has a pool, each area has an independent ventilation system, the specimen preparation area must have an eye wash and a biological safety cabinet, and each area should be equipped with fixed and mobile UV Lights, personnel are assigned to the first area (1), the second area (2), and the third area (1) experimental personnel.

Functions and main equipment of each area of the laboratory




1 Zone 1: Reagent preparation area


       Mainly carry out reagent preparation, sub-packaging and preparation of reaction mixture. The reagent raw materials must be stored in this zone and prepared into the required reagents in this zone. The main equipment in this area includes ultra-clean workbenches, balances, refrigerators, centrifuges, pipettes, shakers, etc. For the control of airflow pressure, there is no strict requirement in this zone, generally +10Pa.




2 Zone 2: Sample Preparation Zone


       Mainly carry out sample preservation, nucleic acid extraction, storage and adding to the amplification reaction reagent. The main equipments in this area include nucleic acid extractor, refrigerator, biological safety cabinet, centrifuge, pipette, shaker, etc. The pressure in this zone is positive relative to zone three to avoid pollution caused by aerosols entering this zone from zone three, generally +10Pa.




3 Three areas: amplification area


       Mainly for DNA amplification. The main equipments in this area are computers and real-time fluorescent quantitative PCR machines. The pressure in this area is negative relative to the adjacent area to prevent aerosol from leaking out of this area, generally -25Pa


#Laboratory ventilation system and pressure control




      In order to prevent the amplification product (aerosol) from entering the pre-amplification area from the three zones along with the air flow, entering each work zone from the corridor should strictly follow a single direction and not travel in the reverse direction. The transfer of reagent samples from each zone must also follow the direction of zone one-zone two-zone three through the transfer window and not flow backwards.


      Each area needs to set up a buffer room, through the installation of exhaust fans, negative pressure exhaust and other devices to make the buffer room form high pressure on the corridor and the work area to ensure that the air in the buffer room flows outwards. When entering the work area from the buffer room, the two doors cannot be opened at the same time.


      In order to avoid the possibility of cross-contamination in various experimental areas, it is advisable to adopt a full-supplied full-exhaust airflow organization form, and strictly control the ratio of supply and exhaust air to ensure the pressure requirements of each experimental area.