Danielson J W, Thompson R D, Bell E
U.S. Food and Drug Administration, Central Laboratory for Microbiological Investigations, Minneapolis, MN 55401, USA.
J AOAC Int. 1999 Jan-Feb;82(1):151-9.
The AOAC sporicidal method uses as a standard the resistance of spores on carriers to 2.5N HCl. This resistance is variable at exposure times ranging from 2 to 20 min. The method described in this paper uses a glutaraldehyde standard and distinguishes various levels of sporicidal activity in the presence of 1-5% glutaraldehyde by using appropriate spore strains, spore preparations, and spore levels. The resistances of 2 Bacillus subtilis 19659 spore preparations cultured in 10% Columbia broth plus manganese and nutrient agar plus minerals, as well as that of B. subtilis var. niger cultured on Lab-Lemco agar, were tested. T-soy broth was a better recovery medium than fluid thioglycollate or modified fluid thioglycollate for B. subtilis 19659 spores exposed to HCl. Sporicidal tests were done on B. subtilis 19659 spores with 2 types of spore preparations. A commercial glutaraldehyde germicide was used for comparison of the sporicidal activity of the glutaraldehyde standard. Two strains of B. subtilis spores and 4 levels of spores (20,000-80,000, 100,000-400,000, 500,000-800,000, and 1,000,000 and up) were removed from check penicylinders from the same batches used for sporicidal tests. B. subtilis var. niger spores were the most resistant to HCl, while B. subtilis 19659 spores were more resistant to glutaraldehyde. Sporicidal activities of a commercial germicide containing 2.5% glutaraldehyde with additives and another containing 5% glutaraldehyde in phosphate buffer were similar. Both totally destroyed high levels of B. subtilis 19659 spores cultured in 10% Columbia broth plus manganese. Results indicate that use of a glutaraldehyde standard, calibrated numbers of spores on penicylinders (bioindicators), and appropriate spore strains and preparations can reduce the variability of sporicidal testing of commercial germicides.
美国官方分析化学师协会（AOAC）的杀孢子方法以载体上的孢子对2.5N盐酸的抗性作为标准。在2至20分钟的暴露时间内，这种抗性是可变的。本文所述方法使用戊二醛标准，并通过使用合适的孢子菌株、孢子制剂和孢子水平，区分在1 - 5%戊二醛存在下的不同杀孢子活性水平。测试了在含锰的10%哥伦比亚肉汤和含矿物质的营养琼脂中培养的2种枯草芽孢杆菌19659孢子制剂以及在Lab - Lemco琼脂上培养的枯草芽孢杆菌黑色变种的抗性。对于暴露于盐酸的枯草芽孢杆菌19659孢子，胰酪大豆肉汤是比硫乙醇酸盐流体培养基或改良硫乙醇酸盐流体培养基更好的复苏培养基。对2种类型孢子制剂的枯草芽孢杆菌19659孢子进行了杀孢子试验。使用一种商业戊二醛杀菌剂来比较戊二醛标准的杀孢子活性。从用于杀孢子试验的同一批次的对照笔式圆筒中取出2株枯草芽孢杆菌孢子和4个孢子水平（20,000 - 80,000、100,000 - 400,000、500,000 - 800,000以及1,000,000及以上）。枯草芽孢杆菌黑色变种孢子对盐酸最具抗性，而枯草芽孢杆菌19659孢子对戊二醛更具抗性。一种含2.5%戊二醛及添加剂的商业杀菌剂和另一种含5%戊二醛的磷酸盐缓冲液杀菌剂的杀孢子活性相似。两者都能完全杀灭在含锰的10%哥伦比亚肉汤中培养的高水平枯草芽孢杆菌19659孢子。结果表明，使用戊二醛标准、笔式圆筒（生物指示剂）上校准数量的孢子以及合适的孢子菌株和制剂，可以降低商业杀菌剂杀孢子测试的变异性。
