Multiplex Real-Time Polymerase Chain Reaction Assay To Detect Acanthamoeba spp., Vermamoeba vermiformis, Naegleria fowleri, and Balamuthia mandrillaris in Different Water Sources.
Elizabeth Córdoba-Lanús, María Reyes-Batlle, Angélica Domínguez-de-Barros, Patricia Pérez-Pérez, Rubén L Rodríguez-Expósito, Alma García-Ramos, Inés Sifaoui, Omar García-Pérez, Germán Aneiros-Giraldez, José E Piñero, Jacob Lorenzo-Morales
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引用次数: 0
Abstract
Free-living amoebae (FLA) are widely distributed in the environment. Among these, Acanthamoeba spp., Naegleria fowleri, Balamuthia mandrillaris, and Vermamoeba vermiformis have been reported as human pathogens with health effects ranging from lethal encephalitis to different epithelial disorders. Despite this, FLA still present many diagnostic challenges. The aim of this study was to develop a rapid and efficient multiplex real-time quantitative polymerase chain reaction (qPCR) to simultaneously detect Acanthamoeba spp., N. fowleri, B. mandrillaris, and V. vermiformis in different water sources. For the validation of the qPCR assay, 38 samples (19 tap water and 19 stagnant water sources) were analyzed. The qPCR assay accurately identified the four types of FLA with no cross-reactivity. Considering water samples with results subsequently confirmed by conventional PCR, the multiplex qPCR assay detected 18/38 (47.4%) positive samples (Acanthamoeba spp. in 44.7% and V. vermiformis in 31.6%) and growth in nonnutritive agar (NNA) cultures identified 7/38 (18.4%) positive samples. Of the tap water samples analyzed, 26.3% of samples positive for FLA were detected by growth in NNA culture whereas 31.6% were identified by qPCR. In addition, FLA were detected in 2/19 stagnant water samples (10.5%) by growth in NNA culture and in 12/19 stagnant water samples (63.2%) by qPCR. Neither N. fowleri nor B. mandrillaris was detected in the water samples analyzed. In conclusion, the qPCR developed showed its potential as a rapid tool for detection of Acanthamoeba spp., N. fowleri, B. mandrillaris, and V. vermiformis. Moreover, FLA species were detected in half of the water sources evaluated, suggesting the importance of the surveillance of these potential infectious agents.
期刊介绍：
The American Journal of Tropical Medicine and Hygiene, established in 1921, is published monthly by the American Society of Tropical Medicine and Hygiene. It is among the top-ranked tropical medicine journals in the world publishing original scientific articles and the latest science covering new research with an emphasis on population, clinical and laboratory science and the application of technology in the fields of tropical medicine, parasitology, immunology, infectious diseases, epidemiology, basic and molecular biology, virology and international medicine.
The Journal publishes unsolicited peer-reviewed manuscripts, review articles, short reports, images in Clinical Tropical Medicine, case studies, reports on the efficacy of new drugs and methods of treatment, prevention and control methodologies,new testing methods and equipment, book reports and Letters to the Editor. Topics range from applied epidemiology in such relevant areas as AIDS to the molecular biology of vaccine development.
The Journal is of interest to epidemiologists, parasitologists, virologists, clinicians, entomologists and public health officials who are concerned with health issues of the tropics, developing nations and emerging infectious diseases. Major granting institutions including philanthropic and governmental institutions active in the public health field, and medical and scientific libraries throughout the world purchase the Journal.
Two or more supplements to the Journal on topics of special interest are published annually. These supplements represent comprehensive and multidisciplinary discussions of issues of concern to tropical disease specialists and health issues of developing countries
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