Comparative analysis of Fluorescence stain and Ziehl-Neelsen's AFB stain for Mycobacterium leprae (pilot study)

Kim, Youn Sil; Lee, Hee Suk; Lee, Min Seok; Kim, Jong Pill

Abstract

The Ziehl-Neelson's AFB staining method was mainly used for the AFB observation of the mycobacteria. However, this method has several issues of false negative results, and hence a comparative experiment of the Ziehl-Neelson's AFB staining and the fluorescence staining method was done to remedy this problem. As the fluorescence staining method brightly highlights the AFB in a dark field, and also as it is observed with the lower power objective, it is a method that can better the observation and shorten the time of observation as well. The fluorescence staining method that was used in this experiment did a comparative analysis of the Auramine O-Rhodamine B and the Acridine Orange. The results showed that although the Auramine O-Rhodamine B allows easier observation of the AFB with a high fluorescence expression rate for the multibacillary leprosy sample, the darkness on the periphery makes it hard to observe anything else, while also making it hard to observe the cell changes and paucibacillary leprosy of the AFB. However, the Acridine Orange staining method highlights the cells in dark green and changes the color of the AFB from bright red to orange making it easier to observe bacilli. The results of the study show that the Acridine Orange method is superior to the Auramine O-Rhodamine B method in detecting acid fast bacilli in specimen.