Diagnosing human cutaneous leishmaniasis using fluorescence in situ hybridization.

Cutaneous leishmaniasis (CL) is endemic in Sri Lanka. Giemsa-stained slit-skin-smears (SSS-Giemsa) and histology are routinely used in diagnosis with a sensitivity of 40-70%. PCR currently has limited accessibility. Therefore, we assessed the sensitivity and specificity of a previously described fluorescence in situ hybridization assay, on skin smears and biopsy samples to overcome the limitations encountered with routine diagnostic methods.Samples from a total of 123 suspected CL patients were collected and subjected to SSS-Giemsa, fluorescence in situ hybridization (FISH) on slit skin smears (SSS-FISH), formalin-fixed-paraffin-embedded-tissues stained with Hematoxylin & Eosin staining (FFPE-H&E) and FISH on formalin-fixed-paraffin-embedded-tissues (FFPE-FISH). Negative controls of 61 patient samples were collected from a CL non-endemic area and subjected to the same procedures. The gold standard PCR was used as a comparator. For FISH, two previously described cyanine 3 tagged Leihsmania genus-specific probes were used.Compared to PCR, SSS-Giemsa, SSS-FISH, FFPE-H&E, and FFPE-FISH had sensitivities of 76.5%, 79.1%, 50.4% and 80.9%, respectively. Routine diagnostic tests (SSS-Giemsa and FFPE-H&E) had a specificity of 100%. SSS-FISH and FFPE-FISH had specificities of 96.7% and 93.4%, respectively. FFPE-FISH had a statistically significant higher diagnostic performance than FFPE-H&E (p < 0.001). The relative performance of SSS-Giemsa, SSS-FISH and FFPE-FISH was similar (p > 0.05 for all comparisons).We conclude that FFPE-FISH is a more accurate diagnostic tool than FFPE-H&E. SSS-FISH did not have an additional advantage over SSS-Giemsa in diagnosis. However, SSS-FISH could be recommended as a minimally invasive method in studies assessing wound healing where immunological probes are used.

Diagnosing Cutaneous leishmaniasis using Fluorescence in Situ Hybridization: the Sri Lankan Perspective.

Cutaneous leishmaniasis (CL) caused by Leishmania donovani MON-37 is becoming a major public health problem in Sri Lanka, with 100 new cases per month being reported in endemic regions. Diagnosis of CL is challenging for several reasons. Due to relative specificity and rapidity we propose Fluorescence in Situ Hybridization as a diagnostic tool for CL. The objective was to evaluate the potential of Fluorescence in Situ Hybridization as a diagnostic method for Cutaneous leishmaniasis in Sri Lanka. Literature on current laboratory tests used to diagnose Cutaneous leishmaniasis in Sri Lanka and globally was reviewed. Sri Lankan data were reviewed systematically following the PRISMA guidelines. A narrative of the results is presented. There is currently no gold standard diagnostic method for Cutaneous leishmaniasis. Fluorescence in Situ Hybridization has been previously applied to detect dermal pathologies including those involving infectious agents, and its use to detect the Leishmania parasite in human cutaneous lesions reported in small number of studies, generally with limited numbers of subjects. Advantages of FISH has been specificity, cost and ease-of-use compared to the alternatives. Based on the available literature and our current work, FISH has potential for diagnosing CL and should now be evaluated in larger cohorts in endemic regions. FISH for CL diagnosis could find application in countries such as Sri Lanka, where laboratory facilities may be limited in rural areas where the disease burden is highest.