What is the role of tissue culture in the workup of leishmaniasis?

Updated: Feb 18, 2020
  • Author: Craig G Stark, MD, FACP, FFTM, RCPS(Glasg), FISTM; Chief Editor: Pranatharthi Haran Chandrasekar, MBBS, MD  more...
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Answer

In vitro cultures of tissue are regularly obtained to aid in leishmaniasis diagnosis and to help identify difficult Leishmania species. This technique has approximately the same diagnostic sensitivity as that of pathologic evaluation, but special laboratory capabilities and technical skills are required. These are available in only 2 reference laboratories in the United States (ie, the Centers for Disease Control and Prevention [CDC], Walter Reed Army Institute of Research).

The value of this method is that the species of the parasite can be identified on the basis of long-standardized isoenzyme patterns on cellulose acetate electrophoresis after the parasite is grown in vitro using both the New World methods of Kreutzer and the Old World methods of Dedet.

Specimens may be cultured on Novy-MacNeal-Nicolle (NNN) medium (rabbit-blood agar that has an overlay of Locke solution with added antibiotics), rabbit blood agar, Schneider Drosophila medium, or a multitude of specialized media to induce promastigote growth. Cultures usually take a few days to 2 weeks to demonstrate growth. Positive culture results occur approximately 75% of the time.

Additional cultures can be performed by inoculating tissue into the footpad and nose of hamsters or certain highly susceptible mouse strains (ie, in vivo cultures via animal inoculation). This is a sensitive method, especially in difficult cases, but results can take several weeks to months.

With successful culture, the parasite can be sent to specialized facilities or the Centers for Disease Control and Prevention (CDC) for polymerase chain reaction (PCR), isoenzyme electrophoresis, or monoclonal antibody speciation.


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