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Leishmaniasis is an important travel-related parasitic infection in the United States. Treatment regimens vary by Leishmania species and require an accurate diagnosis. The sensitivity and specificity of diagnostic methods depend on the type and condition of specimen analyzed. To identify the best algorithm for detection of parasites in fresh and fixed tissue samples, we evaluated parasite cultures, two PCR methods, and Leishmania immunohistochemistry (IHC) in samples received by the CDC from 2012 through 2019. The sensitivity and specificity of IHC assays were evaluated in fresh specimens tested. Diagnostic accuracy for formalin-fixed tissue was evaluated by using PCR-based methods and IHC. Of 100 suspected cases with fresh tissue available, Leishmania spp. infection was identified by PCR in 56% (56/100) of specimens; from these, 80% (45/56) were positive by parasite culture and 59% (33/56) by IHC. Of 420 possible cases where only fixed specimens were available, 58% (244/420) were positive by IHC and/or PCR. Of these, 96% (235/420) were positive by IHC and 84% (204/420) by PCR-based methods. Overall parasite detection using all methodologies was similar for fresh and formalin-fixed tissue specimens (56% versus 58%, respectively). Although PCR-based methods were superior for diagnosis of leishmaniasis and species identification in fresh samples, IHC in combination with PCR increased the accuracy for Leishmania spp. detection in fixed samples. In conclusion, PCR is the most effective method for detecting Leishmania infection in fresh tissue samples, whereas for formalin-fixed samples, IHC and PCR-based methods should be used in combination.
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http://dx.doi.org/10.4269/ajtmh.23-0387 | DOI Listing |
PLoS Negl Trop Dis
September 2025
Department of Clinical Science, Liverpool School of Tropical Medicine, Liverpool, United Kingdom.
Background: Salmonella enterica encompasses over 2,600 serovars, including several commonly associated with severe infection in humans. Salmonella is a major cause of sepsis in Africa; however, diagnosis requires clinical microbiology facilities. Environmental surveillance has the potential to play a role in Salmonella surveillance.
View Article and Find Full Text PDFEur J Haematol
September 2025
Department of Hematology, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
Background: Polymerase chain reaction (PCR)-based Minimal residual disease (MRD) detection is commonly used for core-binding factor acute myeloid leukemia (CBF-AML), but its interpretation in the context of allogeneic hematopoietic stem cell transplantation (allo-HSCT) remains under discussion.
Method: Using Kyoto Stem Cell Transplantation Group registry data, we included 96 patients who underwent allo-HSCT between 2000 and 2019 for CBF-AML.
Results: To assess MRD, quantitative PCR with GAPDH control was most used.
J Microbiol Methods
September 2025
French Armed Forces Medical Directorate, Veterinary Quality Unit, Paris, France.
Foodborne diseases are caused by various pathogens and generally present with similar symptoms, mainly digestive disorders. Adopting a syndromic approach is therefore important when investigating foodborne disease outbreaks. This involves using multiplex PCR-based methods to test stool and food samples.
View Article and Find Full Text PDFmedRxiv
August 2025
Department of Molecular Biology, Princeton University, Princeton, NJ 08544.
Avian influenza viruses (AIVs) are zoonotic pathogens that pose an increasing global threat due to their potential for significant economic losses in agriculture, spillover into humans, and the risk of a pandemic should human-to-human transmission occur. These concerns underscore the need for rapid, sensitive and specific tools to detect and differentiate circulating AIV subtypes and clades. Current AIV diagnostic methods rely on specialized equipment and trained personnel, limiting their use in the field and in low-resource settings.
View Article and Find Full Text PDFAngew Chem Int Ed Engl
September 2025
Beijing Life Science Academy, Beijing, 102206, China.
In-field molecular diagnostics of plant pathogens are critical for crop disease management and precision agriculture, but tools are still lacking. Herein, we present a bioluminescent molecular diagnostic assay capable of detecting viable pathogens directly in minimally processed plant samples, enabling rapid and precise in-field crop disease diagnosis. The assay, called bioluminescent craspase diagnostics (BioCrastics), leverages newly discovered RNA-activated protease of CRISPR (Craspase) with enzymatic luminescence to generate a cascaded amplification, thus bypasses nucleic acid purification and amplification while achieving sub-nanogram sensitivity for fungal pathogens.
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