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Free Content Evaluation of a two-reagent cold staining method for detection of acid-fast bacilli

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SETTING: Tuberculosis Research Centre, Chennai, India.

OBJECTIVE: To evaluate a two-reagent cold staining method for detection of acid-fast bacilli in sputum smears.

SPUTUM SAMPLES: Two hundred and forty-four samples from pulmonary tuberculosis patients attending Tuberculosis Research Centre were used.

METHODS AND DESIGN: Two smears were prepared from each of the samples, of which one was allotted to the two-reagent cold staining method and the other to the Ziehl-Neelsen (Z-N) method. The smears were read blind by a single technician. To ensure correct grading, a senior technician checked all positives and 20% of the negative smears. All the samples were processed by modified PetroffÕs method for culture of Mycobacterium tuberculosis.

RESULTS: The concordance (smear grade one above and one below) between the methods was 90% (kappa value, 0.7). The performance of the cold method and the Z-N method was similar when their smear results were compared with culture results (cold method vs. culture, kappa = 0.61; ZN method vs. culture, kappa = 0.67).

CONCLUSION: The two-reagent cold staining method was found to be as sensitive and specific as the Z-N method. However, large-scale multicentric studies in different climatic conditions need to be conducted to assess its efficacy in the diagnosis of pulmonary tuberculosis.

Keywords: AFB; Ziehl-Neelsen; cold stain; tuberculosis

Document Type: Regular Paper

Affiliations: Tuberculosis Research Centre (ICMR), Chennai, India

Publication date: 2002-08-01

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  • The International Journal of Tuberculosis and Lung Disease publishes articles on all aspects of lung health, including public health-related issues such as training programmes, cost-benefit analysis, legislation, epidemiology, intervention studies and health systems research. The IJTLD is dedicated to the continuing education of physicians and health personnel and the dissemination of information on tuberculosis and lung health world-wide.

    Certain IJTLD articles are selected for translation into French, Spanish, Chinese or Russian. They are available on the Union website

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