Diagnosing human cutaneous leishmaniasis using fluorescence<i>in situ</i>hybridization

Kaluarachchi, T D Jayasena; Wickremasinghe, Rajitha; Weerasekera, Manjula; Yasawardene, S.G.; McBain, Andrew J.; Yapa, Bandujith; Silva, Hiromel De; Menike, Chandranie; Jayathilake, S.; Munasinghe, Anuradha; Wickremasinghe, Renu; Ranasinghe, Shalindra

doi:10.1080/20477724.2021.1896265

Cited by 6 publications

(4 citation statements)

References 16 publications

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“…in FFPE skin samples from human patients using generic probes for Leishmania spp. different from the one used in the present study [12,36]. Frickmann et al [12] detected L. major/tropica amastigotes in four out of eleven (36%) FFPE skin samples from human patients with a clinical diagnosis of old world cutaneous leishmaniasis (OWCL).…”

Section: Discussionmentioning

confidence: 61%

“…Frickmann et al [12] detected L. major/tropica amastigotes in four out of eleven (36%) FFPE skin samples from human patients with a clinical diagnosis of old world cutaneous leishmaniasis (OWCL). On the other hand, Kaluarachchi et al [36], using the same generic probes developed by Frickmann et al [12], identified a higher sensitivity (80.9%), compared with HP (50.4%) for the detection of Leishmania amastigotes in FFPE skins from Sri Lankan patients with OWCL caused by L. donovani. These different sensitivity values found by Frickmann et al [12] and Kaluarachchi et al [36], compared with those of the present study, were probably related to the use of FISH with different probes and the diagnosis of positive skin samples for old world Leishmania species.…”

Section: Discussionmentioning

confidence: 98%

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Comparison between Colorimetric In Situ Hybridization, Histopathology, and Immunohistochemistry for the Diagnosis of New World Cutaneous Leishmaniasis in Human Skin Samples

et al. 2022

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New world cutaneous leishmaniasis (NWCL) is an anthropozoonosis caused by different species of the protozoan Leishmania. Colorimetric in situ hybridization (CISH) was shown to satisfactorily detect amastigote forms of Leishmania spp. in animal tissues, yet it was not tested for the diagnosis of human NWCL. The aim of this study was to compare CISH, histopathology (HP), and immunohistochemistry (IHC) techniques to diagnose NWCL in human cutaneous lesions. The sample comprised fifty formalin-fixed, paraffin-embedded skin biopsy specimens from patients with NWCL caused by L. (V.) braziliensis. These specimens were analyzed by CISH, using a generic probe for Leishmania, IHC, and HP to assess the sensitivity of these methods by using a parasitological culture as a standard reference. Additional specimens from three patients diagnosed with cutaneous mycoses were also included to evaluate cross-reactions between CISH and IHC. The sensitivities of IHC, CISH, and HP for detecting amastigotes was 66%, 54%, and 50%, respectively. IHC, unlike CISH, cross-reacted with different species of fungi. Together, these results demonstrate that CISH may be a complementary assay for the detection of amastigote in the laboratorial diagnosis routine of human NWCL caused by L. (V.) braziliensis.

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Section: Discussionmentioning

confidence: 61%

Section: Discussionmentioning

confidence: 98%

Comparison between Colorimetric In Situ Hybridization, Histopathology, and Immunohistochemistry for the Diagnosis of New World Cutaneous Leishmaniasis in Human Skin Samples

et al. 2022

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“…In 2023, Riyal et al, identified a similar staining pattern in L. donovani causing CL for the first time, with a sensitivity of 74% [ 61 ]. While fluorescence in situ hybridization (FISH) is not a common approach in diagnosing CL, applying Leishmania genus-specific FISH probes on FFPE tissue has been shown to improve sensitivity over conventional staining ( Table 3 ) [ 70 ].…”

Section: Direct Parasitological Methods For Diagnosis Of CLmentioning

confidence: 99%

Diagnostic Tools for Cutaneous Leishmaniasis Caused by Leishmania donovani: A Narrative Review

Piyasiri,

Dewasurendra,

Samaranayake

et al. 2023

Diagnostics

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Leishmaniasis, a neglected tropical disease, encompasses a spectrum of clinical conditions and poses a significant risk of infection to over one billion people worldwide. Visceral leishmaniasis (VL) in the Indian sub-continent (ISC), where the causative parasite is Leishmania donovani, is targeted for elimination by 2025, with some countries already reaching such targets. Other clinical phenotypes due to the same species could act as a reservoir of parasites and thus pose a challenge to successful control and elimination. Sri Lanka has consistently reported cutaneous leishmaniasis (CL) due to L. donovani as the primary disease presentation over several decades. Similar findings of atypical phenotypes of L. donovani have also been reported from several other countries/regions in the Old World. In this review, we discuss the applicability of different methods in diagnosing CL due to L. donovani and a comprehensive assessment of diagnostic methods spanning clinical, microscopic, molecular, and immunological approaches. By incorporating evidence from Sri Lanka and other regions on L. donovani-related CL, we thoroughly evaluate the accuracy, feasibility, and relevance of these diagnostic tools. We also discuss the challenges and complexities linked to diagnosing CL and review novel approaches and their applicability for detecting CL.

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“…Fluorescence in situ hybridization (FISH) allows for the identification and detection of precise DNA sequences or RNA molecules within cells or tissue samples via staining [1,2]. This technique is highly effective in providing researchers with chromosomal and genomic information that aids in the comprehension of gene rearrangements, cytogenetic variation, as well as chromosomal abnormalities and localization.…”

Section: Introductionmentioning

confidence: 99%

An Improved Nested U-Net Network for Fluorescence In Situ Hybridization Cell Image Segmentation

Jian,

Song,

Zhang

et al. 2024

Sensors

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Fluorescence in situ hybridization (FISH) is a powerful cytogenetic method used to precisely detect and localize nucleic acid sequences. This technique is proving to be an invaluable tool in medical diagnostics and has made significant contributions to biology and the life sciences. However, the number of cells is large and the nucleic acid sequences are disorganized in the FISH images taken using the microscope. Processing and analyzing images is a time-consuming and laborious task for researchers, as it can easily tire the human eyes and lead to errors in judgment. In recent years, deep learning has made significant progress in the field of medical imaging, especially the successful application of introducing the attention mechanism. The attention mechanism, as a key component of deep learning, improves the understanding and interpretation of medical images by giving different weights to different regions of the image, enabling the model to focus more on important features. To address the challenges in FISH image analysis, we combined medical imaging with deep learning to develop the SEAM-Unet++ automated cell contour segmentation algorithm with integrated attention mechanism. The significant advantage of this algorithm is that it improves the accuracy of cell contours in FISH images. Experiments have demonstrated that by introducing the attention mechanism, our method is able to segment cells that are adherent to each other more efficiently.

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Diagnosing human cutaneous leishmaniasis using fluorescencein situhybridization

Cited by 6 publications

References 16 publications

Comparison between Colorimetric In Situ Hybridization, Histopathology, and Immunohistochemistry for the Diagnosis of New World Cutaneous Leishmaniasis in Human Skin Samples

Comparison between Colorimetric In Situ Hybridization, Histopathology, and Immunohistochemistry for the Diagnosis of New World Cutaneous Leishmaniasis in Human Skin Samples

Diagnostic Tools for Cutaneous Leishmaniasis Caused by Leishmania donovani: A Narrative Review

An Improved Nested U-Net Network for Fluorescence In Situ Hybridization Cell Image Segmentation

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