Funashima Yumiko, Kato Kyohei, Narita Taeko, Hanaiwa Hiroki, Akiwa Makoto, Sekiguchi Jun-Ichiro, Nagasawa Zenzo, Sugahara Kazuyuki, Miyamoto Hiroshi
Department of Medical Technology and Science, Faculty of Fukuoka Health Care, International University of Health and Welfare.
Department of Clinical Laboratory, Medical Kouhoukai Takagi Hospital.
Rinsho Biseibutshu Jinsoku Shindan Kenkyukai Shi. 2017 Mar 22;27(1):33-41.
Recently, many laboratories use fluorescence microscopy for rapid screening of clinical specimens for detection of Genus . The success of the stain depends on the staining temperature at which the fluorescent dye could uniformly penetrate the cell wall through waxy lipid barrier of the mycobacterial organism. Therefore, this process requires a precise heating control. In this study, to control the temperature during fluorescent auramine- rhodamine staining, we explored the potential use of microwave. The efficiency of microwave irradiation during the staining process was evaluated by using a -containing sputum of which the smear slide was irradiated with several different conditions in combination of time and wattage. As a result, 1) the liquid temperature of the stain correlated well with wattage of microwave irradiation. 2) The tubercle bacilli were easily visualized as brilliant fluorescent bacilli in an orange color when it was set at the best condition of 600 W and 10 sec irradiation. 3) The sensitivity of microscopy with this staining method (MW method) was higher than those of conventional staining methods such as Ziehl-Neelsen staining and standard auramine-rhodamine staining, demonstrating that MW method can be applicable to the sputum slides which contained a few bacilli. Thus, we established the new staining method that is rapid and easy to perform in clinical laboratories. Since the MW method has not yet been utilized in order to conduct fluorescence microscopy for sputum smears, advancement on this method will make a vast change in testing of acid fast bacilli.
最近,许多实验室使用荧光显微镜对临床标本进行快速筛查以检测某属菌。染色的成功取决于染色温度,在该温度下荧光染料能够通过分枝杆菌生物体的蜡质脂质屏障均匀地穿透细胞壁。因此,这个过程需要精确的加热控制。在本研究中,为了在荧光金胺 - 罗丹明染色过程中控制温度,我们探索了微波的潜在用途。通过使用含有的痰液,其涂片载玻片在时间和功率的几种不同条件组合下进行照射,来评估染色过程中微波照射的效率。结果,1)染色液的温度与微波照射功率密切相关。2)当设置为600W和10秒照射的最佳条件时,结核杆菌很容易被视为橙色的明亮荧光杆菌。3)这种染色方法(MW法)的显微镜检查灵敏度高于传统染色方法,如萋 - 尼染色和标准金胺 - 罗丹明染色,表明MW法可适用于含有少量杆菌的痰涂片。因此,我们建立了一种在临床实验室中快速且易于操作的新染色方法。由于MW法尚未用于痰涂片的荧光显微镜检查,该方法的进展将使抗酸杆菌检测发生巨大变化。