疾病预防控制中心实验室建设之PCR 实验室基本要求

疾病预防控制中心实验室建设之PCR 实验室基本要求
1.1.1   PCR 实验室基本要求
2.1  PCR 实验室依据所使用的方法可分为试剂准备、标本制备、扩增、产物分析等三或四个区域。只是使用实时荧光PCR仪、HIV病毒载量测定仪的PCR实验室，有上述前面三个区域即可。各区域应完全独立分隔，不应有任何的空气直通。
Based on the methods used by PCR lab, the PCR laboratory can be divided into three or four rooms or areas for (i) reagent preparation, (ii) sample preparation, (iii) amplification area, and (iv) product analysis area. If the PCR laboratory only uses real-time fluorescence PCR instruments and /or instruments for HIV virus load, the PCR lab may have only the first three rooms or areas (reagent preparation, sample preparation, amplification). The rooms or areas of the PCR lab should be independently separated, and there is no air flow through the separated rooms or areas.
2.2
            产物分析区或三个区域的最后一个区域（即扩增及产物分析区），空气流向应由室外向室内，可通过在室内设置通风橱、排风扇或其他排风系统达到空气流由室外向室内流动的要求。
For PCR product analysis area or last area of three areas (area for amplification and product analysis), direction of air flow shall be from outside to inside of the area by setting up fume hood, exhaust fan or other exhaust system in the area.
2.3 PCR实验室各区域仪器设备配备通常如下：
The equipment configuration for PCR labs are as follows:
2.3.1
            试剂准备区: Reagent preparation area
仪器设备主要应有加样器、冰箱、天平、低速离心机、混匀器、可移动紫外灯等。可使用超净工作台作为试剂配制操作台面。
The main equipment here should be: pipette, refrigerator, balance, low speed centrifuge, vortex mixer, mobile UV lamp, etc. It can use Laminar flow cabinet for the reagent preparation.
2.3.2
            标本制备区sample preparation area
仪器设备主要应有生物安全柜（最好为B2, 可避免提取核酸在柜内反复循环，造成标本间交叉“污染”，出现假阳性结果。此外还应配备加样器、台式高速离心机（冷冻及常温）、台式低速离心机、恒温设备（水浴和/或干浴仪）、冰箱、混匀器和可移动紫外灯等。
The main equipment is the bio-safety cabinet (type B2 is better, because it can avoid recycling of nucleic acid cabinet and cross contamination of specimens). Besides, it shall be equipped with pipettes, centrifuge table high speed (refrigerated and normal temperature), centrifuge table low speed, constant temperature devices (water bath or dry bath), refrigerator, vortex mixer and mobile lamp UV etc.
2.3.3
            扩增区amplification area
主要仪器就是核酸扩增热循环仪（PCR仪，实时荧光或普通的）。热循环仪的电源应专用，并配备一个稳压电源或UPS，以防止由于电压的波动对扩增测定的影响。此外，根据工作需要，还可配备加样器、超净台等。
The main equipment here is a thermal cycler system for nucleic acid amplification (PCR, quantitative fluorescence detection or common). Power for the thermal cycler shall be provided with a manostat or a UPS to avoid the influence on PCR amplification arose by fluctuation of voltage. Moreover, depending on the work demand, the amplification area could be equipped with pipette, ultra clean cabinet, etc.
2.3.4
            产物分析区：PCR product analysis area
本区所使用的仪器设备可能有加样器、电泳仪（槽）、电转印仪、杂交炉或杂交箱、水浴箱、DNA测序仪、酶标仪和洗板机等。
The equipment here may be pipette, electrophoresis system capillary, transblot system, chest/ oven molecule hybridization, water bath, DNA sequencer, microplate-reader for ELISA, microplate washer and so on.
建议Recommendations
在理想情况下，PCR实验室试剂准备、标本制备、扩增三个区域可设置缓冲间。在缓冲间内，可设置正压，使室内空气不流向室外，室外空气不流向室内。
In an optimal situation, for three rooms (reagent preparation, sample preparation, amplification) of the PCR lab one buffer area could be set up. The pressure in the buffer area should be positive to assure the indoor air could not flow to outside and the outside air could not flow to the inside.