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首页> 外文期刊>感染症学雑誌 >Evaluation of an improved pretreatment method for the measurement of (1-->3)-beta-D-glucan in blood samples
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Evaluation of an improved pretreatment method for the measurement of (1-->3)-beta-D-glucan in blood samples

机译：评价一种改进的预处理方法以测量血液样品中的（1-> 3）-β-D-葡聚糖

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    Measurement of blood (1-->3)-beta-D-glucan is useful for early diagnosis and follow-up of the therapeutic process of deep seated mycoses. The Fungitec G test MK (Seikagaku Corp., Tokyo) kit using alkaline-pretreatment followed by chromogenic kinetic assay has been widely used in Japan because of its high sensitivity and easy handling of a large number of samples. Discrepancy in the levels of (1-->3)-beta-D-glucan and/or in the quantitative judgement, however, has been pointed out between this kit and other commercial kits. One of the reasons for this discrepancy has been reported to be non-specific reactions caused by substances other than beta-glucan. In this study, we have improved the alkaline pretreatment reagent by changing the concentration of KOH and salts, resulting in a marked reduction of the non-specific reaction. Recovery of standard beta-glucan added to plasma or serum after the improved pretreatment was 80 to 120%, and no amidolytic activity was detected either in plasma or in serum. By the improved pretreatment, the incidence of non-specific reactions, i.e., those that exceed the quantitation limit (3.9 pg/mL), were markedly decreased from 139 to 16 out of 200 plasma samples and from 106 to 22 out of 170 serum samples. The incidence of strong non-specific reactions, i.e., those that exceed the cut-off level (20 pg/mL), were also decreased from seven to one with plasma and seven to zero with serum samples. Correlation between corrected beta-glucan measurements by the current pretreatment and non-corrected ones by the improved pretreatment was quite good. The improved method is thus expected to decrease the frequency of non-specific false-positive reactions, with the high sensitivity of Fungitec G test MK.
    机译：血液（1-> 3）-β-D-葡聚糖的测量可用于深层真菌病的早期诊断和治疗过程的随访。在日本，使用碱预处理并随后进行生色动力学测定的Fungitec G测试MK（东京精工制药公司）试剂盒由于其高灵敏度和易于处理大量样品而被广泛使用。但是，已经指出了该试剂盒与其他商用试剂盒之间（1-> 3）-β-D-葡聚糖的水平和/或定量判断上的差异。据报道，造成这种差异的原因之一是除β-葡聚糖以外的物质引起的非特异性反应。在这项研究中，我们通过改变KOH和盐的浓度改进了碱性预处理试剂，从而显着减少了非特异性反应。经过改进的预处理后，添加到血浆或血清中的标准β-葡聚糖的回收率为80％至120％，并且血浆或血清中均未检测到酰胺分解活性。通过改进的预处理，非特异性反应（即超过定量限（3.9 pg / mL）的反应）的发生率显着降低，从200个血浆样品中的139个降至16个，从170个血清样品中的106个降至22个。强烈的非特异性反应（即超过临界水平（20 pg / mL）的反应）的发生率也从血浆中的7种降至1种，血清样品中的7种降至零。当前预处理的校正后β-葡聚糖测量值与改进的预处理后的未校正测量值之间的相关性非常好。因此，改进的方法有望以Fungitec G测试MK的高灵敏度降低非特异性假阳性反应的频率。

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