1. LAMP kit for diagnosis of non-falciparum malaria in Plasmodium ovale infected patients
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José M. Rubio, Gerardo Rojo-Marcos, Xavier C. Ding, Iveth J. González, Peña Gómez-Herruz, Alexandra Martin Ramírez, Ramon Perez Tanoira, Juan Cuadros, Federal Ministry of Education & Research (Alemania), KfW Development Bank, Clinicum, Department of Ophthalmology and Otorhinolaryngology, and HUS Head and Neck Center
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medicine.medical_specialty ,030231 tropical medicine ,Plasmodium ovale ,Loop-mediated isothermal amplification ,Biology ,Sensitivity and Specificity ,03 medical and health sciences ,0302 clinical medicine ,Predictive Value of Tests ,LAMP ,Positive predicative value ,parasitic diseases ,medicine ,Humans ,MEDIATED ISOTHERMAL AMPLIFICATION ,ASSAY ,030212 general & internal medicine ,ELIMINATION ,Research ,medicine.disease ,biology.organism_classification ,Virology ,3. Good health ,Malaria ,Europe ,Diagnosis of malaria ,Infectious Diseases ,Molecular Diagnostic Techniques ,Parasitology ,3121 General medicine, internal medicine and other clinical medicine ,Tropical medicine ,Malaria control ,Nucleic Acid Amplification Techniques - Abstract
BACKGROUND: Microscopy and rapid diagnosis tests have a limited sensitivity in diagnosis of malaria by Plasmodium ovale. The LAMP kit (LoopAMP®) can be used in the field without special equipment and could have an important role in malaria control programmes in endemic areas and for malaria diagnosis in returned travellers. The performance of the Pan primer of the kit in detecting malaria by P. ovale was compared with the results of standard nPCR in samples of patients returning from P. ovale endemic areas. METHODS: Plasmodium ovale positive samples (29, tested by PCR and/or microscopy) and malaria negative specimens (398, tested by microscopy and PCR) were collected in different hospitals of Europe from June 2014 to March 2016 and frozen at -20 °C. Boil and spin method was used to extract DNA from all samples and amplification was performed with LoopAMP® MALARIA kit (Eiken Chemical, Japan) in an automated turbidimeter (Eiken 500). The results of LAMP read by turbidimetry and with the naked eye were compared. RESULTS: The kit showed a sensitivity of 100% and a specificity of 97.24% with positive and negative predictive values of 72.5 and 100%, respectively. Naked eyed readings were in accordance with turbidimetry readings (sensitivity, 92.5%, specificity, 98.96% and positive and negative predictive values, respectively, 90.24 and 99.22%). The limit of detection of LAMP assay for P. ovale was between 0.8 and 2 parasites/µl. CONCLUSIONS: The Pan primer of the Malaria kit LoopAMP® can detect P. ovale at very low-levels and showed a predictive negative value of 100%. This tool can be useful in malaria control and elimination programmes and in returned travellers from P. ovale endemic areas. Naked eye readings are equivalent to automated turbidimeter readings in specimens obtained with EDTA. This work received financial support from FIND with funds from the German Federal Ministry of Education and Research (BMBF) through the KfW Entwicklungsbank. Sí
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