Assessment of pan-Leishmania detection by recombinase polymerase amplification assay |
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| Affiliation: | 1. Institute for Medical Microbiology and Virology, University Medical Center Goettingen, Georg-August University, Göttingen, Germany;2. Institute of Animal Hygiene and Veterinary Public Health, University of Leipzig, Leipzig, Germany;3. Nutrition and Clinical Services Division, International Centre for Diarrheal Disease Research Bangladesh, Dhaka, Bangladesh;4. Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Pengkalan Chepa, Kota Baru, Kelantan, Malaysia;5. Faculty of Medical Sciences, University of Sri Jayewardenepura, Gangodawila, Nugegoda, Sri Lanka;6. WHO Collaborating Center for Leishmaniasis, National Center for Microbiology, Instituto de Salud Carlos III, Madrid, Spain;8. Laboratory Sciences and Services Division, International Centre for Diarrheal Disease Research Bangladesh, Dhaka, Bangladesh;9. Department of Dermatology, Venereology and Allergology, University Medical Centre Göttingen, Georg-August University, Göttingen, Germany;1. Laboratory of Human and Experimental Pathology, Pasteur Institute of Tunis, Tunis, Tunisia;2. Laboratory of Molecular Epidemiology and Experimental Pathology, Pasteur Institute of Tunis, Tunis, Tunisia;3. High Institute of Sciences and Technology of Environments of Borj-Cedria, University of Carthage, Tunis, Tunisia;1. Laboratoire de Parasitologie-Mycologie, CHU Nîmes, Nîmes, France;2. Laboratoire de Parasitologie-Mycologie, CHU Nîmes, Université de Montpellier, CNRS, IRD, MiVEGEC, Montpellier, France;1. Department of Parasitology, Faculty of Medicine, University of Colombo, Colombo, Sri Lanka;2. Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Colombo, Colombo, Sri Lanka;3. Department of Pharmacology, Institute of Postgraduate Medical Education and Research, Kolkata, West Bengal, India;1. Departamento de Parasitologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, Brazil.;2. Departamento de Biologia Animal, Instituto de Biologia, Universidade Estadual de Campinas, Campinas, Brazil;3. Departamento de Biologia, Facultad de Ciencias Básicas, Universidad del Tolima, Tolima, Colombia;1. Laboratoire Hospitalier Universitaire de Bruxelles, (LHUB-ULB) Department of Microbiology, Université Libre de Bruxelles, Brussels, Belgium;2. Department of Gastroenterology and Hepatology, Centre Hospitalier Universitaire Saint-Pierre, Université Libre de Bruxelles, Brussels, Belgium;3. Department of Gastroenterology, Hepatopancreatology and Digestive Oncology, Erasme University Hospital, Université Libre de Bruxelles, Brussels, Belgium;4. Gastroenterology Unit, Centre Hospitalier Universitaire Brugmann, Université Libre de Bruxelles, Brussels, Belgium;5. Gastroenterology Department, Centre Hospitalier Inter régional Edith Cavell, sites de la Basilique et E. Cavell, Brussels, Belgium;6. Gastroenterology Unit, Hôpital Universitaire des Enfants Reine Fabiola, Université Libre de Bruxelles, Brussels, Belgium;7. Belgian Helicobacter and Microbiota Study Group (BHMSG), Brussels, Belgium |
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| Abstract: | The spread of vector habitats along with increasing human mobility can introduce atypical Leishmania species and hence can challenge existing diagnostic practices for rapid detection of active infection with species outside the narrow target range. Here we assessed the pan-Leishmania detection ability of isothermal recombinase polymerase amplification (RPA) assays targeting 18S rRNA gene, cathepsin L-like cysteine proteinase B (Cpb) gene, and kinetoplast minicircle DNA (kDNA) regions. While the lowest limit of detection of the 18S rRNA-RPA and Cpb-RPA assays were estimated as 12 and 17 standard DNA molecules, respectively, both assays could amplify genomic DNA of 7 pathogenic Leishmania species. Evaluation of 18S rRNA-RPA and our previously developed kDNA-RPA assays on 70 real-time PCR-positive leishmaniasis samples of varying pathologies resulted in sensitivity rates of 35.71% and 88.57%, respectively, while the combined sensitivity was 98.57%. Combinatorial application of 18S rRNA-RPA and kDNA-RPA assays can be recommended for further diagnostic assessments. |
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