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Lessons From a Proficiency Testing Event for Acid-Fast Microscopy*
Objectives: To evaluate the routine performance and the technical parameters of different acid-fast staining methods: Kinyoun, Ziehl-Neelsen (ZN), auramine, and auramine-rhodamine.
Design and participants: The performance of 167 laboratories was analyzed using prestained and unstained slides.
Setting: Laboratories holding New York State permits.
Results: The results revealed that Kinyoun's cold carbol fuchsin method is inferior to both the ZN and fluorochrome (auramine and/or auramine-rhodamine) methods. Even though 91% of the participants used commercial staining kits, the study identified unexpected errors concerning the concentration of carbol fuchsin, time for staining and counterstaining, and the concentration of acid alcohol for decolorization, which may significantly influence the sensitivity. Besides these findings, the present study showed that the examination of < 300 view fields may also decrease the sensitivity of acid-fast microscopy. In addition, we found that the sensitivity and specificity of the ZN and fluorochrome methods are comparable if the procedural standards are followed.
Conclusions: The strict and ongoing quality control of the "simple to perform" acid-fast microscopy and the immediate review of commercially available staining kits are necessary. Because of the rapidity of the fluorochrome method, laboratories with large specimen numbers should use this technique. In all other cases, the ZN method should be used. Moreover, all clinicians should be aware of the method of acid-fast microscopy used and the proficiency of the laboratory in performing the assay. (CHEST 2001; 120:250-257)
Key words: fluorochrome; Kinyoun; microscopy; quality control; tuberculosis; Ziehl-Neelsen
Abbreviations: AFB = acid-fast bacilli; CDC = Centers for Disease Control and Prevention; CMPH = Clinical Microbiology Procedures Handbook; ZN = Ziehl-Neelsen
Acid-fast microscopy is the fastest, easiest, and least expensive tool for the rapid identification of potentially infectious tuberculosis patients.1,2 The importance of the method is further exemplified by the fact that in low-income countries, the definitive diagnosis of tuberculosis still relies solely on the presence of acid-fast bacilli (AFB) in stained smears.
Although the specificity of acid-fast microscopy is excellent for mycobacterial species, the sensitivity is not optimal. The sensitivity of microscopy is influenced by numerous factors, such as the prevalence and severity of disease, the type of specimen, the quality of specimen collection, the number of mycobacteria present in the...