Abstract
Ancylostoma caninum is a zoonotic nematode which is able to affect animals and humans. Diagnosis in the definitive host and environmental detection are key to prevent its dissemination and achieve control. Herein, a new coprological LAMP method for the detection of A. caninum (Copro-LAMPAc) DNA was developed. DNA extraction was performed using a low-cost method and a fragment of the cox-1 gene was used for primer design. The analytical sensitivity, evaluated with serial dilutions of genomic DNA from A. caninum adult worms, was 100 fg. A specificity of 100% was obtained using genomic DNA from the host and other pathogens. The Copro-LAMPAc was evaluated using environmental canine fecal samples. When compared with gold standard optical microscopy in epidemiological studies, it proved to be more sensitive. This new LAMP assay can provide an alternative protocol for screening and identification of A. caninum for epidemiological studies in endemic areas.
Highlights
Over one-third of the worldwide population harbors a parasitic helminth; these parasitic diseases generate millions of deaths each year [1] and produce a series of morbidities that affect mostly vulnerable populations from low and medium income countries (LMIC), causing millions of disability-adjusted life years (DALYs) [2]
Other conditions occasionally attributed to A. caninum are myositis, unilateral subacute neuroretinitis, and eosinophilic enteritis [3]
Few cases were reported where A. caninum was able to complete its migration to the human intestine [7], generating several eosinophilic gastroenteritis [8]
Summary
Over one-third of the worldwide population harbors a parasitic helminth; these parasitic diseases generate millions of deaths each year [1] and produce a series of morbidities that affect mostly vulnerable populations from low and medium income countries (LMIC), causing millions of disability-adjusted life years (DALYs) [2]. LAMP reactions have been shown to be useful for the detection of pathogens in food and surveillance of water quality [50,51,52,53,54,55,56,57] This technique is not currently widely used in the diagnostic routine, probably due to the high cost of the visualization methods required and the cost of DNA extraction. In this work the development of an easy copro-LAMP reaction for A. caninum detection was performed This reaction was evaluated using two different methods for DNA extraction in order to be able to implement it in laboratories without sophisticated equipment
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