Analytical Performance of a Loop-Mediated Isothermal Amplification Assay for Leishmania DNA Detection in Sandflies and Direct Smears of Patients with Cutaneous Leishmaniasis
| Autor: | Martha S. Ayala, Carolina Flórez, Juan Hember Tabares, Cielo León, Marina Muñoz, Carolina Hernández, Juan David Ramírez |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
RNA 18S Limit of detection Phlebotominae Real time polymerase chain reaction Procedures Diagnostic accuracy law.invention Loop mediated isothermal amplification 0302 clinical medicine law Nucleic acid amplification Leishmaniasis Polymerase chain reaction Leishmania Microscopy biology Diagnostic test accuracy study Articles Infectious Diseases Real-time polymerase chain reaction Protozoan Nucleic Acid Amplification Techniques Human Smear Protozoal DNA 030231 tropical medicine 030106 microbiology Loop-mediated isothermal amplification Leishmaniasis Cutaneous DNA determination Colombia Sensitivity and Specificity Article Skin leishmaniasis 03 medical and health sciences Cutaneous leishmaniasis Virology parasitic diseases Genetics medicine Animals Humans Detection limit Animal DNA Nucleic acid amplification technique DNA Protozoan Nonhuman biology.organism_classification medicine.disease Molecular biology Cutaneous Isolation and purification Parasitology Psychodidae |
| Zdroj: | The American Journal of Tropical Medicine and Hygiene Repositorio EdocUR-U. Rosario Universidad del Rosario instacron:Universidad del Rosario |
| Popis: | Loop-mediated isothermal amplification (LAMP) is ideal for the detection of Leishmania DNA as it is a quick and easy-to-perform test that does not require complex or sophisticated equipment or infrastructure. However, the application of this technique in the detection of Leishmania DNA has not been comprehensively analyzed to date (analytical validation). Our objective was to evaluate the sensitivity and analytical specificity (anticipated reportable range [ARR], the limit of detection [LoD], and accuracy) of LAMP targeting the 18S rRNA gene in the diagnosis of six New World Leishmania species. We then applied the validated LAMP assay across 50 samples of sandflies and 50 direct smears from a recent outbreak of cutaneous leishmaniasis in Colombia to determine its diagnostic performance. The LAMP assay exclusively amplified the DNA of Leishmania spp., and an ARR of between 1 × 10 4 and 1 × 10 -2 equivalent parasites/mL was determined. An LoD of 1 × 10 -2 equivalent parasites/mL was established and there was no statistically significant variation in terms of accuracy. Finally, a sensitivity of 100% in direct smears and sandflies samples was calculated and a specificity of 90.9% for direct smears using microscopy as reference and 96.8% for sandflies using real-time polymerase chain reaction as reference were determined. To our knowledge, this is the first attempt to analytically validate a LAMP test to detect Leishmania DNA, which showed good diagnostic potential from sandflies and direct smear samples. Copyright © 2018 by The American Society of Tropical Medicine and Hygiene. |
| Databáze: | OpenAIRE |
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