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疾控中心實驗室設計之PCR 實驗室基本要求
2023-07-05 10:49 來源:

疾控中心實驗室設計之PCR 實驗室基本要求


1.1.1  PCR實驗室設計規劃的基本要求

  2.1 PCR 實驗室依據所使用的方法可分為試劑準備、標本製備、擴增、產物分析等三或四個區域。隻是使用實時熒光PCR儀、HIV病毒載量測定儀的PCR實驗室,有上述前麵三個區域即可。各區域應完全獨立分隔,不應有任何的空氣直通。

  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 產物分析區或三個區域的最後一個區域(即擴增及產物分析區),空氣流向應由室外向室內,可通過在室內設置通風櫥、排風扇或其他排風係統達到空氣流由室外向室內流動的要求。

  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實驗室各區域儀器設備配備通常如下:

  The equipment configuration for PCR labs are as follows:

  2.3 .1試劑準備區: Reagent preparation area

  儀器設備主要應有加樣器、冰箱、天平、低速離心機、混勻器、可移動紫外燈等。可使用超淨工作台作為試劑配製操作台麵。

  The main equipment here should be: pipette, refrigerator, balance, low speed centrifuge, vortex mixer, mobiles UV lamp, etc. It can use Laminar flow cabinet for the reagent preparation.

  2.3 .2標本製備區sample preparation area

  儀器設備主要應有生物安全櫃(最好為B2, 可避免提取核酸在櫃內反複循環,造成標本間交叉“汙染”,出現假陽性結果。此外還應配備加樣器、台式高速離心機(冷凍及常溫)、台式低速離心機、恒溫設備(水浴和/或幹浴儀)、冰箱、混勻器和可移動紫外燈等。

  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 mobiles lamp UV etc.

  2.3 .3擴增區amplification area

  主要儀器就是核酸擴增熱循環儀(PCR儀,實時熒光或普通的)。熱循環儀的電源應專用,並配備一個穩壓電源或UPS,以防止由於電壓的波動對擴增測定的影響。此外,根據工作需要,還可配備加樣器、超淨台等。

  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產物分析區:PCR product analysis area


  本區所使用的儀器設備可能有加樣器、電泳儀(槽)、電轉印儀、雜交爐或雜交箱、水浴箱、DNA測序儀、酶標儀和洗板機等。

  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.


  建議Recommendations

  在理想情況下,PCR實驗室試劑準備、標本製備、擴增三個區域可設置緩衝間。在緩衝間內,可設置正壓,使室內空氣不流向室外,室外空氣不流向室內。

  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.



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