Leishmania Diagnostics Laboratory

collage showing leishmania wound, computer with data, a graph, and cells

The WRAIR's Leishmania Diagnostics Laboratory (LDL) in Silver Spring, MD, was established in 1978 and was accredited in 2001 by the College of American Pathologists (CAP) as a response to growing outbreaks of leishmaniasis during Operations Iraqi Freedom and Enduring Freedom where it became a significant source of morbidity for U.S. Service Members. Transmitted by infected sand flies, leishmaniasis remains a meaningful threat to the Warfighter.

LDL is the only accredited leishmaniasis reference diagnostic laboratory in North America. LDL provides Leishmania testing services to U.S. Service Members, their beneficiaries and DOD civilian employees located in the continental United States (CONUS) and outside the continental United States (OCONUS). Tests offered include: microscopic evaluation of histopathology slides, Leishmania culture, real-time PCR, antibody detection (rK39) and species determination (cellulose acetate electrophoresis: CAE).

CONUS Testing Services

CONUS Specimen Submission Form: please fill the request form completely and mail to the listed address for timely specimen processing.

Culture:

The diagnosis of leishmaniasis is made by microscopic identification of the in-vitro culture of the motile, extracellular form (promastigote) on suitable media provided by LDL. 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 (CAE) after sufficient parasite material is grown in-vitro.

Specimen Types Accepted:
          o    Dermal Scrapings
          o    Punch Biopsy
          o    Needle Aspirates
          o    Bone Marrow

Specimen Requirements:
          o    Medium: Specimen can be placed in LDL Schneider’s medium (provided upon request), Phosphate Buffered Saline or RPMI
          o    Shipping Conditions: Ambient 15-30°C

Histopathology:

 The diagnosis of leishmaniasis is made by microscopic identification of the nonmotile, intracellular form (amastigote) in stained sections from lesions, or thin touch prep.

Specimen Types Accepted:
          o    Dermal Scrapings
          o    Touch Prep-Impression Smears (Slides)

Specimen Requirements:
          o    Microscope slides: Fixed or unfixed Dermal Scrapings or Touch prep-impression smears
          o    Shipping Conditions: unfixed, alcohol fixed or stained slides can be shipped at Ambient temperature 15-30°C

Molecular Testing:

SMART Leish is an FDA-approved qualitative diagnostic real-time PCR test for the rapid detection of Leishmania species from individuals suspected of having leishmaniasis (510K: 081868). Any tissue, obtained by any method (aspirate, scraping, biopsy) can be placed into a screw cap tube of alcohol (ethanol, isopropanol) and then sent for PCR analysis. 

Specimen Types Accepted:
          o    Dermal Scrapings
          o    Punch Biopsy
          o    Needle Aspirates
          o    Bone Marrow (VL)

Specimen Requirements:
          o    Any tissue, obtained by any method (aspirate, scraping, biopsy) can be placed into a tube of ethanol and then sent for PCR analysis.
          o    Shipping Conditions: Ambient 15-30°C

Serological Testing:

The Kalazar Detect™ Rapid Test for Visceral Leishmaniasis (VL) is a rapid immunochromatographic strip assay for the qualitative detection of antibodies to members of L. donovani in human serum to aid in the presumptive diagnosis of VL. Antibodies against this antigen are present in serum/blood of VL patients and they bind to the recombinant antigen that is bound to the test strip yielding a visual binary reading (positive or negative) test result.

Specimen Types Accepted:
          o    Serum

Specimen Requirements:
          o    1-2 mL serum
          o    Draw Tube/Medium: SST Tubes or Red-top tube
          o    Shipping Conditions: 2-8 °C post-centrifugation, shipped in cold box with ice packs

OCONUS Testing Services

OCONUS Specimen Submission Form: please fill the request form completely and mail to the listed address for timely specimen processing.

Histopathology:

The diagnosis of leishmaniasis is made by microscopic identification of the nonmotile, intracellular form (amastigote) in stained sections from lesions, or thin touch prep.

Specimen Types Accepted:
          o    Touch prep-impression smears of biopsies on dermal scrapings

Specimen Requirements:
          o    Microscope slides: Fixed or unfixed Dermal Scrapings or Touch prep-impression smears
          o    Shipping Conditions: unfixed, alcohol fixed or stained slides can be shipped at Ambient temperature 15-30°C

Molecular Testing:

SMART Leish is an FDA-approved qualitative diagnostic real-time PCR test for the rapid detection of Leishmania species from individuals suspected of having leishmaniasis (510K: 081868). Any tissue, obtained by any method (aspirate, scraping, biopsy, formalin-fixed) can be placed into a screw cap tube of alcohol (ethanol, isopropanol) and then sent for PCR analysis. 

Specimen Types Accepted:
          o    Dermal Scrapings
          o    Punch Biopsy

Specimen Requirements:
          o    Any tissue, obtained by any method (aspirate, scraping, biopsy) can be placed into a tube of ethanol and then sent for PCR analysis.
          o    Shipping Conditions: Ambient 15-30°C

Serological Testing:

The Kalazar Detect™ Rapid Test for Visceral Leishmaniasis (VL) is a rapid immunochromatographic strip assay for the qualitative detection of antibodies to members of L. donovani in human serum to aid in the presumptive diagnosis of VL. Antibodies against this antigen are present in serum/blood of VL patients and they bind to the recombinant antigen that is bound to the test strip yielding a visual binary reading (positive or negative) test result.

Specimen Types Accepted:
          o    Serum

Specimen Requirements:
          o    1-2mL serum
          o    Draw Tube/Medium: SST Tubes or Red-top tube
          o    Shipping Conditions: 2-8°C post-centrifugation, shipped in cold box with ice packs

Contact Us!

Please contact us with any questions via LDL’s email or telephone.

24 Hour Phone: 240-595-7353
Email: usarmy.detrick.medcom-wrair.mbx.leishmania-diagnostic@mail.mil

Please send slides and cultures via overnight shipping to the following address:
Leishmania Diagnostic Laboratory Department of Laboratory Diagnostics and Monitoring US HIV Military Research Program, Walter Reed Army Institute of Research ATTN: Leishmania Diagnostics Laboratory (LDL)
9100 Brookville Road, Building 508, Silver Spring, MD 20910

References:     

Diagnosis and Treatment of Leishmaniasis: Clinical Practice Guidelines by the Infectious Diseases Society of America (IDSA) and the American Society of Tropical Medicine and Hygiene (ASTMH). Clin Infect Dis. 2016 Dec 15;63(12):e202-e264. Epub 2016 Nov 14