Diagnostic performance of Ziehl-Neelsen staining and Auramine-Rhodamine staining techniques in the detection of pulmonary and extrapulmonary tuberculosis

Gustavo Tapia-Sequeiros; Miguel Hueda-Zavaleta; Juan Carlos Gómez de la Torre; Arantsa Hernandez-Vargas; Claudia Barletta-Carrillo; Cinthya Flores; Cristian Piscoche; Cecilia Miranda; Ada Mendoza

doi:10.17843/rpmesp.2025.421.14062

Authors

Gustavo Tapia-Sequeiros Facultad de Ciencias de la Salud, Universidad Privada de Tacna, Tacna, Perú. Diagnóstico, tratamiento e investigación de enfermedades infecciosas y tropicales, Universidad Privada de Tacna, Tacna, Perú. https://orcid.org/0000-0002-9277-2731
Miguel Hueda-Zavaleta Diagnóstico, tratamiento e investigación de enfermedades infecciosas y tropicales, Universidad Privada de Tacna, Tacna, Perú. Hospital III Daniel Alcides Carrión – Essalud, Calana, Tacna, Perú. https://orcid.org/0000-0002-8049-7787
Juan Carlos Gómez de la Torre Sequence Reference Lab, San Isidro, Lima, Perú. https://orcid.org/0000-0003-4566-2027
Arantsa Hernandez-Vargas Facultad de Ciencias de la Salud, Universidad Privada de Tacna, Tacna, Perú. https://orcid.org/0000-0002-3637-7139
Claudia Barletta-Carrillo Sequence Reference Lab, San Isidro, Lima, Perú. https://orcid.org/0000-0001-9982-5368
Cinthya Flores Sequence Reference Lab, San Isidro, Lima, Perú. https://orcid.org/0009-0009-3452-3276
Cristian Piscoche Sequence Reference Lab, San Isidro, Lima, Perú https://orcid.org/0000-0002-9907-8013
Cecilia Miranda Sequence Reference Lab, San Isidro, Lima, Perú https://orcid.org/0009-0000-7904-5735
Ada Mendoza Sequence Reference Lab, San Isidro, Lima, Perú https://orcid.org/0009-0008-7202-856X

DOI:

https://doi.org/10.17843/rpmesp.2025.421.14062

Keywords:

Tuberculosis, Mycobacterium tuberculosis, Microbiological Techniques, Rapid Diagnostic Tests, Peru

Abstract

Objective. To determine the diagnostic performance of Ziehl-Neelsen and Auramine-Rhodamine staining for pulmonary and extrapulmonary tuberculosis. Materials and methods. This was a cross-sectional diagnostic test study. We used a database of processed samples from patients with suspected pulmonary and extrapulmonary tuberculosis in a private laboratory in Peru between 2011 and 2022. Ziehl-Neelsen staining and Auramine-Rhodamine staining were the index tests. The reference tests were Xpert MTB/RIF and Xpert Ultra. The receiver operating characteristics and area under the curve (AUC/ROC) were calculated to determine the diagnostic performance of each staining technique. Results. We analyzed 908 samples processed by Ziehl-Neelsen staining and 623 samples by Auramine-Rhodamine staining, most were lung tissue samples. Using the Xpert MTB/RIF as a reference test, the Ziehl-Neelsen staining obtained an AUC=0.72, a sensitivity of 44.2% and specificity of 99.3%; and Auramine-Rhodamine staining showed an AUC=0.73, a sensitivity of 46.8% and specificity of 100%. Considering the Xpert Ultra test as a reference, the diagnostic performance for Ziehl-Neelsen showed an AUC=0.63 with a sensitivity of 26.9% and specificity of 98.5%; and an AUC=0.64 with a sensitivity of 30% and specificity of 98.2% for the Auramine-Rhodamine staining. Conclusion. The diagnostic performance for both staining techniques is fair, and decreases when taking Xpert Ultra as a reference. New diagnostic alternatives with adequate performance are
needed to detect tuberculosis.

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