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首页> 外文期刊>Journal of Clinical Microbiology >Turbidimetric method for quantifying serum inhibition of Limulus amoebocyte lysate.
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Turbidimetric method for quantifying serum inhibition of Limulus amoebocyte lysate.

机译：浊度法定量定量Li变形细胞裂解液的血清抑制作用。

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This study describes a method to quantify the inhibition of lipopolysaccharide (LPS) activity by serum with a turbidimetric Limulus amoebocyte lysate assay. Assays were performed in multiwell microplates, and turbidity was measured as the optical density at 380 nm with a microplate spectrophotometer. LPS potency was measured as the 50% maximal Limulus amoebocyte response (LR50) of LPS diluted with saline. By comparing LR50s in saline, LPSs from various species of bacteria were standardized against the U.S. Reference Standard Endotoxin, lot EC-5. The potency of Escherichia coli O113 and O18 and Serratia marcescens LPSs was found to be equal to that of the reference standard EC-5, whereas LPSs from two salmonella species were half as potent. The least potent LPSs tested, obtained from Klebsiella pneumoniae and E. coli rough mutant J5, were 5- and 10-fold less potent, respectively, than EC-5. As a measure of inhibition, the LR50 of LPS in serum was compared to the LR50 of LPS in saline. Serum inhibited the potency of LPS 103- to 6,400-fold compared with saline. A positive correlation was found between standardized potency in saline and serum inhibition of the various LPSs tested. Thus, LPSs from E. coli O113, O18, and EC-5 and S. marcescens, which exhibited the highest potency in saline, were inhibited the most by serum. Likewise, E. coli J5 and K. pneumoniae LPSs, which were the least potent tested, were the least inhibited. The degree of inhibition of all types of LPS tested increased with increasing serum concentration.
机译：这项研究描述了一种通过比浊Li变形细胞溶血产物测定来量化血清对脂多糖（LPS）活性抑制的方法。在多孔微孔板中进行测定，并用微孔板分光光度计在380 nm处以光密度测量浊度。以用盐水稀释的LPS的50％最大Li变形细胞反应（LR50）来测量LPS效能。通过比较盐水中的LR50，针对美国参考标准内毒素，批次EC-5对来自各种细菌的LPS进行了标准化。发现大肠杆菌O113和O18和粘质沙雷氏菌LPS的功效与参考标准EC-5相同，而来自两种沙门氏菌的LPS的功效却是其一半。从肺炎克雷伯菌和大肠杆菌粗糙突变体J5获得的最低效LPS的效价分别比EC-5低5倍和10倍。作为抑制的量度，将血清中LPS的LR50与盐水中LPS的LR50进行比较。与盐水相比，血清将LPS的效力抑制了103倍至6,400倍。在盐水中的标准化效价与测试的各种LPS的血清抑制之间发现正相关。因此，血清中对大肠杆菌O113，O18，EC-5和marcescens的LPS表现出最高的生理盐水效力，它们对盐水的抑制作用最大。同样，对大肠杆菌J5和肺炎克雷伯氏菌LPS的抑制作用最弱，其测试效果最低。所测试的所有LPS类型的抑制程度都随着血清浓度的增加而增加。

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