发光二极管荧光显微镜在基层实验室检测分枝杆菌的价值

Application of light-emitting diode fluorescence microscopy in detection of Mycobacterium in county level laboratories

目的研究发光二极管(I-ED)荧光显微镜检测分枝杆菌在基层实验室的应用价值。方法连续纳入2017年7月至2018年1()月黑龙江省8个县级结核病防治机构的初诊肺结核可疑症状者,共3770例.每例患者留取2~3份痰标本.供分析的痰标本共9079份.全部进行了痰结核分枝杆菌固体培养。由实验室人员取每份痰标本制备2张痰涂片.分别进行萋尔-尼尔逊染色(简称“萋尼染色”)和荧光染色.然后使用普通光学显微镜和LED荧光显微镜观察结果?同时对阅片时间及涂片保存进行记录和分析.结果萋-尼染色普通显微镜和荧光染色LED荧光显微镜镜检痰涂片阳性率分别为11.41%(430/3770)和12.79%(482/3770);痰固体培养阳性率为17.14%(646/3770),差异有统计学意义(x2=5602.10.P<0.01)。以痰培养结果为参照标准.萋-尼染色普通显微镜和荧光染色I.EI)荧光显微镜镜检痰涂片分枝杆菌的敏感度分别为61.46%(397,646)和66.72%(431/646).特异度分别为9&94%(3091/3124)和9&37%(3073/3124);萋-尼染色普通显微镜镜检约登指数为0.604,荧光染色LED荧光显微镜镜检约登指数为0.651。LED荧光显微镜镜检痰涂片所用时间为(184.33±52.22)s.明显低于普通光学显微镜镜检所用时间[(291.21±95.40)s].差异有统计学意义(F=5670.80.PV0.01)。LED荧光显微镜检测的1179张阳性级别不同的荧光染色痰涂片放入不透光的玻片盒内常温(22~25°C)存放.并避免阳光直射.存放4个月发现有5张定性错误.10张定量误差。结论I.EI)荧光显微镜检査抗酸杆菌的检测效能优于普通光学显微镜,缩短了阅片时间,染色的玻片在常温避免阳光直射的玻片盒内保存即可,适合在基层实验室推广。

Oljjective To study the practicability of light-cmitting diode ( LED) fluorescence microscope in laboratory staff to examine Mycobacterium. Methods A total of 3770 newly diagnosed TB suspected patients were continuously included from eight county-level TB dispensaries of Heilongjiang from July 2017 to October 2018. Each patient provided 2 -3 sputum specimens* and 9079 examples were analyzed and cultured. The laboratory staff prepared two sputum smears for each sample. Ziel-Nelson (Z-N) and fluorescence staining for the smear were conducted, respectively. Then traditional light microscopes and LED fluorescence microscopes were used for micro scopic examination. At the same lime, reading time and smear preservation were ret'orded and analyzed. Results The positive rates of traditional light microscopy and LED fluorescence microscopy were 11.41%(430/3770) and 12. 79%(482/3770);the positive detection rate of simple nlethod of solid culture was 17. 14%(646,, 3770). the difference was statistically significani (才=5602. 1()? P<C0. 01). Using culture test as the standard, the sensitivity of microscopy by traditional light microscope and LEI) fluorescence microscope was 61. 46%(397/646) and 66. 72%(431 646), and the specificity was 9& 94%(3091/3124) and 98. 37%(3073/3124). respectively. Youden index of traditional light microscope and LED fluorescence microscope was 0. 604 and 0. 651. The reading time of LED fluo rescence microscope ((184. 33士52. 22) s) was obviously lower than that of traditional light microscope ((291. 21 ± 95. 40) s), and the difference was statistically significant (F=5670. 80, PVO. 01). 1179 positive fluorescent stai ning smears with different positive grades detected by LEI) fluorescence microscopy were putted in the opaque glass box at room temperature (22 - 25 °C ), and avoided direct sunlight. Five qualitative errors and 10 quantitative errors were found after 4 months of storage. Conclusion The efficiency of LED fluorescence microscopy in detecting acid-fast bacilli is better than that of ordinary optical microscopy. It shortened the film reading time of the staff. The stained glass slides can be stored in the glass box that avoids direct sunlight at room temperature, which is suitable for promotion and application in the basic laboratory.