Phenol-Free Staining Kit Detects Mycobacteria

A safe, clean, and user-friendly staining kit has been developed for fluorescence detection of mycobacteria in histologic and clinical material.

Up to now, phenol was the first choice among additives to staining solutions. But because of its numerous noxious qualities, safety measures had to be taken during its application. The new, phenol-free staining kit prevents harm to the user's health and provides clean results.

Mycobacteria are difficult to stain because of the high proportion of lipid and wax in their cell walls. The stained acid fast mycobacteria keep their coloring even after treatment with strong decolorizing solutions as hydrochloric acid (HCl) ethanol. The Merck KGaA (Darmstadt, Germany) kit uses a modified staining solution that makes inclusion of phenol in the staining solution unnecessary.

The Tb-fluor phenol-free is suitable for paraffin sections and clinical specimen such as sputum, bronchial lavage, and urine. The sensitivity and specificity of the staining results are identical with those obtained using the classical (phenol-employing) staining method.

Liquid material can be pre-treated with Sputofluol to oxidatively free the mycobacteria from the surrounding cell and mucous material by the action of hypochlorit, an ingredient of Sputofluol. The kit contains auramine O and rhodamine B in the staining solution for more flexibility and optical brilliance. A positive result is visible with red-orange or yellow-green fluorescence.

To be able to gain an optimal impression of the fluorescence, the intrinsic fluorescence of the background is suppressed with potassium permanganate.


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