Optimizing the Glass Bead Sterilization Protocol Focusing on Removal of Organic and Bacterial Intraoperative Contamination.

Keith Lewy, Jonathan Bova, Timothy A Erickson, Robert Rose, Sara D Lawhon, Tracy H Vemulapalli
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Abstract

Validated glass bead sterilization protocols to effectively sterilize rodent surgical instruments after bacterial exposure (for example, cecal contamination) are lacking. To refine current approaches, we added either a multienzyme detergent, neutral pH detergent, or chlorhexidine scrub step before glass bead sterilization of forceps or needle drivers exposed to cecal contents. We exposed sets of forceps and needle drivers to cecal contents, which were then air dried for 3 min. Immediately after, the instruments were wiped several times with a clean, dry paper towel. The contaminated tips were soaked in either a multienzyme or neutral pH detergent (t = 5 min), chlorhexidine scrub (t = 2 min), or no pretreatment solution. To further increase debris removal, instruments (from all groups) were brushed using a clean toothbrush. The nonpretreatment instruments were briefly soaked in saline before brushing. After being rinsed with sterile water, all instruments were exposed to a glass bead sterilizer for 60 s at 500 °F (260 °C). Sets were then swabbed for bacterial culturing. Swabs were plated onto either sheep blood agar (n = 23) or chocolate agar (n = 20) for aerobic culturing or Brucella agar (n = 20) for anaerobic culturing. A subset of instruments was sampled to determine organic material presence after treatment using an ATP luminometer (n = 21). Multiple agar types and bioluminescence were used to more deeply evaluate tool sterility and to differentiate the relative effectiveness of each protocol. From the saline group, only one pair of forceps yielded growth on Brucella agar, and 2 pairs yielded growth on chocolate agar. No other bacterial growth was observed. The use of a pretreatment agent also lowered overall organic contamination levels in needle drivers compared with using only saline. These results indicate that brushing instruments to mechanically remove debris from instruments is paramount to ensure sterility. However, a best practice would be to also use one of the pretreatment options used in this study.

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优化玻璃微珠灭菌方案,重点清除术中有机物和细菌污染。
目前还缺乏经过验证的玻璃珠灭菌方案,无法在啮齿动物外科器械接触细菌(例如盲肠污染)后对其进行有效灭菌。为了改进目前的方法,我们在对暴露于盲肠内容物的镊子或针头驱动装置进行玻璃珠灭菌之前,添加了多酶洗涤剂、中性 pH 值洗涤剂或洗必泰擦洗步骤。我们将成套的镊子和针刀暴露于盲肠内容物中,然后风干 3 分钟。然后立即用干净的干纸巾擦拭器械数次。将受污染的针尖浸泡在多酶或中性 pH 值洗涤剂(t = 5 分钟)、洗必泰擦洗液(t = 2 分钟)或无预处理溶液中。为了进一步清除碎屑,使用干净的牙刷刷洗器械(所有组别)。未进行预处理的器械在刷牙前在生理盐水中短暂浸泡。用无菌水冲洗后,将所有器械放在玻璃珠灭菌器中以 500 °F(260 °C)的温度灭菌 60 秒。然后用拭子进行细菌培养。将拭子滴在羊血琼脂(n = 23)或巧克力琼脂(n = 20)上进行需氧培养,或滴在布鲁氏菌琼脂(n = 20)上进行厌氧培养。使用 ATP 发光器(n = 21)对一部分仪器进行取样,以确定处理后是否存在有机物质。使用多种琼脂类型和生物发光法更深入地评估工具的无菌性,并区分每种方案的相对效果。在生理盐水组中,只有一对镊子在布鲁氏菌琼脂上生长,两对在巧克力琼脂上生长。没有观察到其他细菌生长。与仅使用生理盐水相比,使用预处理剂也降低了针头驱动装置中有机物的总体污染水平。这些结果表明,刷洗器械以机械方式清除器械上的碎屑对确保无菌至关重要。不过,最佳做法是同时使用本研究中使用的预处理方案之一。
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