【正文】 entration, it may be necessary to modify the dilution series and to use more plates at each dilution. Prepare a separate series of plates for each aliquot. Place mL of each selected dilution in each of two 15 100mm Petri dishes. Within 20 minutes, add to each plate 20 mL of Soybean–Casein Digest Agar Medium (see Microbial Limit Tests 61 ) that has been melted and cooled to 45 to 50 . Swirl to attain a homogeneous suspension, and allow to solidify. 如果此生物指示劑的孢子濃度較低，則有必要改變稀釋系列，在每個(gè)稀釋級(jí)增加更多的平皿。在 0℃～4 ℃冰浴中迅速冷卻。For Biological Indicator for Steam Sterilization, Paper Carrier, heat the tube containing the suspension in a water bath at 95 to 100 for 15 minutes (heat shock), starting the timing when the temperature reaches 95 . 如果是紙載的蒸汽滅菌生物指示劑，則把裝有懸浮液的管放入 95℃～100℃水浴15min（熱休克），在懸浮液溫度達(dá)到 95℃時(shí)開(kāi)始計(jì)時(shí)。把杯子里的紙片磨碎成纖維狀，并混合 3～5min 使之成為均一的懸浮液。USP 55 BIOLOGICAL INDICATORS—RESISTANCE PERFORMANCE TESTS 生物指示劑－抗性測(cè)試TOTAL VIABLE SPORE COUNT 活孢子數(shù)總數(shù)計(jì)數(shù)Remove three specimens of the relevant biological indicator from their original individual containers. Disperse the paper into ponent fibers by placing the test specimens in a sterile 250mL cup of a suitable blender containing 100 mL of chilled, sterilized Purified Water and blending for 3 to 5 minutes to achieve a homogeneous suspension. Transfer a 10mL aliquot of the suspension to a sterile, screwcapped 16 125mm tube. 從生物指示劑的原包裝袋中取出 3 支作為樣本。把待測(cè)樣品放入一個(gè)已滅菌的 250mL 有攪拌器的杯子里，杯子內(nèi)裝 100 mL 放冷的滅菌純化水。轉(zhuǎn)移 10ml 的整數(shù)倍的懸浮液至一個(gè)已滅菌的16 125mm 的螺蓋管中。For Biological Indicator for DryHeat Sterilization, Paper Carrier, and for Biological Indicator for Ethylene Oxide Sterilization, Paper Carrier, heat the tube containing the suspension in a water bath at 80 to 85 for 10 minutes, starting the timing when the temperature reaches 80 . Cool rapidly in an ice water bath at 0 to 4 . 如果是紙載的干熱滅菌生物指示劑及環(huán)氧乙烷滅菌生物指示劑，則把裝有懸浮液的管放入 80℃～85℃水浴 10min，在懸浮液溫度達(dá)到 80℃時(shí)開(kāi)始計(jì)時(shí)。Transfer two 1mL aliquots to suitable tubes, and make appropriate serial dilutions in sterilized Purified Water, the dilutions being selected as calculated to yield preferably 30 to 300 colonies, but not less than 6, on each of a pair of plates when treated as described below. 轉(zhuǎn)移 2 個(gè) 1ml 的懸浮液到合適的試管，用滅菌純化水進(jìn)行適宜的系列稀釋，所選稀釋液要求在進(jìn)行下述操作后能達(dá)到 30～300CFU ／皿，但不少于 6 CFU／皿。每一取整數(shù)倍制備一個(gè)單獨(dú)的系列平皿。在 20min 內(nèi)，向每個(gè)平皿中倒入 20 mL 溶解并冷卻到45℃～50℃的大豆－酪蛋白消化物瓊脂（參考 Microbial Limit Tests 61 ）。Incubate the plates in an inverted position at 55 to 60 for Biological Indicator for Steam Sterilization, Paper Carrier, and at 30 to 35 for Biological Indicator for Ethylene Oxide Sterilization, Paper Carrier, and for Biological Indicator for DryHeat Sterilization, Paper Carrier, or at the optimal recovery temperature specified by the manufacturer, and examine the plates after 24 and 48 hours, recording for each plate the number of colonies, and using the number of colonies after 48 hours to calculate the results. 倒扣平皿進(jìn)行培養(yǎng)，紙載蒸汽滅菌生物指示劑：55℃～60 ℃培養(yǎng)，紙載的干熱滅菌生物指示劑及環(huán)氧乙烷滅菌生物指示劑：30 ℃～35 ℃培養(yǎng)，或在供應(yīng)商指定的有最佳回收率的溫度下培養(yǎng)，24h 和 48h 后檢查平皿，記錄各皿的菌落數(shù)，用 48h 后的菌落數(shù)進(jìn)行結(jié)果計(jì)算。如果每個(gè)懸浮液 48h 的孢子數(shù)的對(duì)數(shù)等于或大于其 24h 的孢子數(shù)的對(duì)數(shù)，則實(shí)驗(yàn)成立。D VALUE DETERMINATION D 值的確定For all tests described in this section, handle each test specimen with aseptic precautions, using sterilized equipment where applicable.在本部分的所有實(shí)驗(yàn)中，要在無(wú)菌的條件下處理各檢測(cè)樣品，并使用無(wú)菌的器具。The geometrical pattern of the heat source(s) is such as to enable the biological indicators under test to be uniformly heated under the specified conditions. The temperature profile in the chamber is known, and cold spots, hot spots, and slow heat zones identified. 熱源的幾何模式是用來(lái)確保受測(cè)的生物指示劑能在特定的條件下受熱均勻。The chamber has the capability to work within a temperature range of 40 to 300 , with an accuracy at any particular setting of not less than 177。2℃。For Biological Indicator for Ethylene Oxide Sterilization, Paper Carrier, use an apparatus that consists of a test chamber with a means of ensuring adequate mixing of the sterilant gas and a means of heating the sterilant gas to not lower than the preselected operating temperature so that no liquid enters the test chamber, equipped with temperature control and monitoring, pressure control, humidification, and gas concentration monitoring devices. 對(duì)于紙載環(huán)氧乙烷滅菌生物指示劑，使用帶有測(cè)試腔的設(shè)備，此測(cè)試腔可以確保滅菌氣體充分混勻，并且可以把滅菌氣體加熱到不低于預(yù)設(shè)的運(yùn)行溫度從而沒(méi)有液體進(jìn)入這個(gè)測(cè)試腔。Detailed specifications and operational parameters for suitable apparatus are those published in Standard for a Biological Indicator—Evaluator Resistometer for Ethylene Oxide Gas Vessels (BIER/EO) Gas 設(shè)備的詳細(xì)說(shuō)明和和運(yùn)行參數(shù)發(fā)表載在《生物指示劑標(biāo)準(zhǔn)－環(huán)氧乙烷氣體管道(BIER/EO)氣體管道鎳鉻表評(píng)估》。設(shè)備的詳細(xì)說(shuō)明和和運(yùn)行參數(shù)發(fā)表載在《生物指示劑標(biāo)準(zhǔn)－飽和蒸汽 (BIER/蒸汽管道)鎳鉻表評(píng)估》。從生物指示劑的原包裝盒里取出足量的一組生物指示劑樣本，每組包含 5～10 個(gè)樣本。把每一組置于一個(gè)適宜的樣本容器中，使滅菌腔中特定位置的每個(gè)樣本都能暴露于以上所述的滅菌條件。選擇一系列以樣本檢測(cè)用最短時(shí)間為基礎(chǔ)不斷增加的滅菌時(shí)間。For Biological Indicator for DryHeat Sterilization, Paper Carrier, preheat the sterilizing chamber for 30 minutes. Open the access door or port, place one of the holders with a group of specimens in the sterilizing chamber, close the access door or port, and continue to