Haishima Y, Tanamoto K
Department of Microbiology, National Institute of Hygienic Sciences, Tokyo, Japan.
Microbiol Immunol. 1993;37(10):813-6. doi: 10.1111/j.1348-0421.1993.tb01710.x.
Quantitative microanalysis of bacterial endotoxin was performed using [3H]-labeled L-glycero-D-mannoheptitol (LD-Heptitol) as a marker. Several different amounts of authentic L-glycero-D-mannoheptose (LD-Heptose) were reduced with 20 micrograms of cold NaBH4 containing 2 micrograms of NaB3H4 (40 Ci/mmol) in 20 microliters of 1 mM NaOH at 4 C for 48 hr. The product, [1-3H]-labeled LD-Heptitol, has high specific activity, and was purified by HPLC and detected using a liquid-scintillation counter. As little as 50 pg of LD-Heptose was detectable, and the radioactivity increased dose-dependently in the 100 pg to 80 ng range tested. More than 2 ng of Salmonella abortus equi endotoxin could be accurately determined by this method. It is possible to detect 50 pg of endotoxin by this method, if 100% hot material (NaB3H4) is used for [3H]-labeling.
使用[3H]标记的L-甘油-D-甘露庚糖醇（LD-庚糖醇）作为标志物对细菌内毒素进行定量微量分析。在20微升1 mM氢氧化钠中，于4℃用含有2微克NaB3H4（40 Ci/mmol）的20微克冷NaBH4还原几种不同量的纯L-甘油-D-甘露庚糖（LD-庚糖）48小时。产物[1-3H]标记的LD-庚糖醇具有高比活性，通过高效液相色谱法纯化并用液体闪烁计数器检测。可检测到低至50皮克的LD-庚糖，在所测试的100皮克至80纳克范围内，放射性呈剂量依赖性增加。用此方法可准确测定超过2纳克的马流产沙门氏菌内毒素。如果将100%的热物质（NaB3H4）用于[3H]标记，用此方法可检测到50皮克的内毒素。
