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Rapid Detection of Gram-Negative Bacteriuria by Use of the Limulus Endotoxin Assay

机译：Li内毒素测定法快速检测革兰氏阴性菌尿

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摘要

The Limulus in vitro endotoxin assay was evaluated as a possible method for the prompt detection of significant gram-negative bacteriuria in children. This assay is capable of detecting endotoxin associated with intact cell walls of viable gram-negative bacteria as well as free endotoxin. Quantitative results are obtained following a 1-h incubation of Limulus lysate and 10-fold dilutions of otherwise untreated urine. A standard curve of Limulus activity and viable cell counts of Escherichia coli and Klebsiella pneumoniae in urine demonstrated that a positive Limulus reaction at a dilution of 1:100 or 1:1,000 indicated a colony count of at least 100,000 bacteria/ml. A positive Limulus reaction only from undiluted urine or at a dilution of 1:10 indicated less than 100,000 cells/ml. These experimental observations were confirmed by comparing the Limulus test with quantitative plate counts on 209 urine specimens from a mixed pediatric population. These results indicate that the Limulus assay is a simple, accurate method for rapid presumptive detection of gram-negative bacteriuria in patients where an immediate diagnosis is needed. This test would also seem promising for screening large patient populations for bacteriuria or for monitoring the effectiveness of treatment of urinary tract infections.
机译：Li体外内毒素测定被评估为快速检测儿童重大革兰氏阴性菌尿的一种可能方法。该测定法能够检测与存活的革兰氏阴性细菌的完整细胞壁相关的内毒素以及游离内毒素。将Li裂解物孵育1小时并将未经处理的尿液稀释10倍后即可获得定量结果。尿液中大肠杆菌和肺炎克雷伯菌的Li活性和活细胞计数的标准曲线表明，在1：100或1：1,000的稀释度下，阳性Li反应表明菌落数至少为100,000细菌/ ml。仅未稀释尿液或以1:10稀释液产生的阳性Li反应表明少于100,000个细胞/ ml。通过比较Li试验和定量板计数对混合儿科人群的209个尿液样本，证实了这些实验观察结果。这些结果表明，在需要立即诊断的患者中，Li测定法是一种快速，准确推定革兰氏阴性菌尿的简单，准确的方法。对于筛查大量患者的细菌尿症或监测尿路感染治疗的有效性，该测试似乎也很有希望。

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