[利用磁珠技术检测抗酸杆菌的创新检测方法——结核微珠法的评价]。

Masanori Ohkuma, Katsuyoshi Ikeda, Konen Obayashi, Yukio Ando, Toyoyasu Koriyama, Minhi Kimu, Nobuyuki Hirose, Zenzo Nagasawa, Hiroshi Miyamoto
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引用次数: 0

摘要

使用半碱性蛋白酶(SAP)和NALC-NaOH的离心方法被“2007版结核分枝杆菌检测试验指南”推荐,对提高抗酸杆菌(AFB)的涂片和培养试验的敏感性和特异性做出了重大贡献。然而,这种方法在其繁琐和耗时的分析方案方面提出了一些挑战。“TB-beads (Kyokuto Pharmaceutical Industrial Co., Ltd)”是一种利用磁珠检测AFB的新方法。我们将该方法的质量与离心法进行比较,重点关注涂片和培养试验的结果。本评价研究采用5份阳性和5份阴性痰液样本进行。使用“丙烯酸染色剂”进行的结核菌珠荧光涂片试验的灵敏度几乎与离心机方法相同。然而,结核微珠的一个优点是,由于涂片的玻璃载玻片背景清晰,因此非常方便进行显微观察。两种方法的污染率比较表明,结核微珠的污染率明显较低。分离法对HK半固体分离培养基和BacT/ALERT MP的污染率分别为50%和60%。另一方面,两种培养方法的结核微珠污染率均为10%。对5例阳性痰液,采用Myco Acid、Ogawa K、BacT/ALERT MP对离心机和TB-Beads法的检出率进行比较。TB-Beads法对Myco Acid和Ogawa K的检出率较高,而两种方法对BacT/ALERT MP的检出率无显著差异(16-23 d)。结核菌珠是一种简便的方法,可简化涂片试验过程,并有助于显著降低培养试验的污染率。有助于提高AFB检测的灵敏度和检出率。此外,它不需要离心。最终,TB-beads是一种创新、安全、方便的AFB检测方法,可使实验室技术人员节省日常工作时间。
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[Evaluation of TB-beads assay utilizing the technique of magnetic beads--an innovative assay method for detection of acid fast bacilli].

The centrifuge method with the use of Semi-Alkalin Proteinase (SAP) and NALC-NaOH, recommended by the "2007 edition of the assay guideline for detection of Mycobacterium tuberculosis," has significantly contributed to improving the sensitivities and specificities of both smear and culture tests for detection of acid fast bacilli (AFB). However, this method poses some challenges in terms of its cumbersome and time-consuming assay protocol. "TB-beads (Kyokuto Pharmaceutical Industrial Co., Ltd.)" is a newly-developed method for detection of AFB utilizing magnetic beads. We evaluated the quality of this method in comparison with the centrifuge method, focusing on the results of smear and culture tests. This evaluation study was conducted using both 5 positive and 5 negative sputum samples. The sensitivity of TB-beads for fluorescent smear tests, conducted using "Acri-stain," was almost the same as that of the centrifuge method. One advantage of TB-beads, however, was that it was very convenient to practice microscopic observation due to the clear background of the smeared glass slides. The comparison of the contamination rates between the two methods showed that TB-beads suggested significantly lower contamination rates. The centrifuge method resulted in 50% and 60% of contamination rates for HK Semisolid Isolation Medium and BacT/ALERT MP, respectively. On the other hand, the contamination rates of TB-beads for both of the culture methods were only 10%. With regard to the 5 positive sputum samples, the comparison of the detection rates between the centrifuge and TB-Beads method was made utilizing Myco Acid, Ogawa K, and BacT/ALERT MP. The TB-Beads method suggested higher detection rates for Myco Acid and Ogawa K, while there were no significant differences between the two methods for BacT/ALERT MP (16-23 days). TB-beads is an easy method that allows to simplify the process of smear tests, and contributes to significantly reducing the contamination rate of culture tests. It also contributes to improving the sensitivity and detection rate of AFB testing. Furthermore, it does not require centrifugation. Ultimately, TB-beads is an innovative, safe, and convenient testing method for detection of AFB, which enables laboratory technicians to save time for routine work.

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