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      成品預裝培養基平皿質量要求

      發布時間:

      2022-12-27

      作者:


      成品預裝培養基平皿質量要求
       
       
      當今科學技術飛速發展,有關微生物檢測的高科技方法可謂如若瀚海。雖然培養皿檢測微生物是一個古老的、傳統的檢測方法,但由于它具有可以直接分離鑒別樣品、操作方便、結果可靠等優點,所以至今仍廣泛應用于醫藥衛生,臨床檢驗、實驗動物、食品、化妝品、工農業、環保等眾多領域。在藥品質量控制、安全評價和疾病診斷中尤其發揮著重要的作用。
       
      培養基是微生物試驗的基礎,直接影響微生物試驗結果,而成品預裝培養基平皿是培養基中的一種,是由脫水培養基完全溶解于水中,矯正PH值,然后滅菌和分裝于平皿而成。無論是實驗室配制的培養基平皿或商品化的成品預裝培養基平皿,其質量都依賴于其制備的整個過程。采用不適宜方法制備的培養基將影響微生物的生長或復蘇,從而影響試驗結果的可靠性。所以要確保培養基平皿的質量,需特別注意以下事項:
      1
      選擇適宜的培養基制備方法,不得使用結塊或顏色發生改變的脫水培養基。
      2
      培養基滅菌應按照生產商提供的或經驗證的參數進行,避免采用不適當的加熱和滅菌條件,而引起培養基顏色變化、透明度降低、瓊脂凝固力或pH值的改變。
      3
      應確定每批培養基滅菌后的pH值(冷卻至室溫25℃測定)。若培養基處方中未列出pH值的范圍,除非經驗證表明培養基的pH值允許的變化范圍很寬,否則,pH值的范圍不能超過規定值±0.2。
      4
      成品預裝培養基平皿應確保無菌,并且平皿不得破裂,盡量避免形成氣泡,固體培養基表面不得產生裂縫或漣漪,在冷藏溫度下不得形成結晶,不得污染微生物等。
      5
      培養基升溫滅菌過程中溫度上升緩慢或滅菌后降溫過慢,可能導致培養基的過熱或過度滅菌,一定程度上會降低微生物培養基促生長的質量,所以成品預裝培養基平皿應通過微生物促生長試驗進行驗證。
      6
      用于環境監控的培養基平皿必須特別防護,最好采用三層包裝和終端滅菌。不建議采用實驗室自制的培養皿通過100%預培養后用于潔凈區環境采樣,因為預培養過程中失水會影響微生物生長,且還存在培養皿被污染的可能。
      7
      貯藏和運輸條件應使成品培養基最低限度的失去水分并提供機械保護、避光保存。含瓊脂培養基平皿不得在0℃或0℃以下存放,冷凍可能破壞凝膠特性。
       
      培養基平皿制備過程質量控制和貯藏條件是提供優質培養基的保證。潔凈區環境浮游菌、沉降菌及表面微生物監測用培養基一般采用胰酪大豆胨瓊脂培養基(TSA),必要時可加入適宜的中和劑;當監測結果有疑似真菌或考慮季節因素影響時,可增加沙氏葡萄糖瓊脂培養基(SDA)。
      以用于潔凈環境的微生物監控與微生物檢測的成品培養基和2015版《中華人民共和國藥典》規定純化水微生物限度檢測采用R2A瓊脂培養基為參考,列舉相應質量要求如下表所示:
       
      TSA、SDA、R2A成品預裝培養基平皿質量要求
       
       

      TSA

      SDA

      R2A

      外觀

      表面濕潤,平整、干凈,培養基顏色為淺黃色透明固體,無破損、氣泡、脫離現象。

      pH值

      pH值7.3±0.2

      pH值5.6±0.2

      pH值7.2±0.2

      無菌性

      檢查

      培養5天后應無菌落生長

      培養7天后應無菌落生長

      培養5天后應無菌落生長

      填充量

      Φ90mm培養基的裝量為20ml; Φ55mm的接觸碟裝量應保證瓊脂在培養皿上形成凸起彎月面。

      注意:為避免平皿在取樣過程中失水過多影響微生物生長,可根據潔凈區環境條件選擇適宜的裝量。

      微生物

      促生長

      試驗

      50%≤(銅綠假單胞菌、金黃色葡萄球菌、枯草芽孢桿菌、白色念珠菌、黑曲霉)回收率≤200%,菌落形態、大小與對照板一致

      50%≤(白色念珠菌、黑曲霉)回收率≤200%,菌落形態、大小與對照板一致

      50%≤(銅綠假單胞菌、枯草芽孢桿菌)回收率≤200%,菌落形態、大小與對照板一致

        • 培養皿圖片

       

       

      Quality requirements for finished product preloaded medium plate
      With the rapid development of science and technology, the high technology of microbial detection is like the sea. Although Petri dish detection microorganism is an ancient and traditional detection method, it has been widely used in many fields, such as medical hygiene, clinical test, laboratory animal, food, cosmetics, industry and agriculture, environmental protection and so on, because it can separate and distinguish samples directly, and it has the advantages of convenient operation and reliable results. It plays an important role in drug quality control, safety evaluation and disease diagnosis.
      The culture medium is the basis of microbiological test, which directly affects the results of microbiological test. The product preloaded medium plate is one of the medium, which is completely dissolved in water, corrected the pH value, and then sterilized and distributed in a Petri dish. Whether the laboratory preparation of the culture medium or the commercialized finished product, the quality of the dish is dependent on the whole process of its preparation. The culture medium prepared by unsuitable method will affect the growth or recovery of microorganisms, thus affecting the reliability of the test results. Therefore, in order to ensure the quality of the dishes, we need to pay special attention to the following:
      One
      Choose suitable medium preparation method, and do not use dehydrated medium with caking or color changing.
      Two
      Culture medium sterilization should be carried out in accordance with the manufacturer's or verified parameters to avoid improper heating and sterilization conditions, resulting in changes in the color change of the medium, the decrease of transparency, the agar agar or the pH value.
      Three
      The pH value of each batch of medium after sterilization should be determined (cooling to room temperature at 25
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