Detection of Schistosoma mansoni Eggs in Feces through their Interaction with Paramagnetic Beads in a Magnetic Field

Abstract

Diagnosis of intestinal schistosomiasis in low endemic areas is a problem because often control measures have reduced egg burdens in feces to below the detection limits of classical coproparasitological methods. Evaluation of molecular methods is hindered by the absence of an established standard with maximum sensitivity and specificity. One strategy to optimize method performance, where eggs are rare events, is to examine large amounts of feces. A novel diagnostic method for isolation of Schistosoma mansoni eggs in feces, and an initial evaluation of its performance is reported here. Known amounts of S. mansoni eggs were seeded into 30 g of normal human feces and subjected to a sequence of spontaneous sedimentation, sieving, Ritchie method, incubation and isolation through interaction with paramagnetic beads. Preliminary tests demonstrated the efficacy of lectins as ligands, but they also indicated that the paramagnetic beads alone were sufficient to isolate the eggs under a magnetic field through an unknown mechanism. Eggs were identified by microscopic inspection, with a sensitivity of 100% at 1.3 eggs per gram of feces (epg). Sensitivity gradually decreased to 25% at a concentration of 0.1 epg. In a preliminary application of the new method to the investigation of a recently established focus in southern Brazil, approximately 3 times more eggs were detected than with the thick-smear Kato-Katz method. The novel S. mansoni detection method may significantly improve diagnosis of infections with low burdens in areas of recent introduction of the parasite, areas under successful control of transmission, or in infected travelers. It may also improve the evaluation of new treatments and vaccines. Schistosomiasis mansoni is a parasitic infection that affects approximately 200 million people, mainly in the tropics. The worms live inside the veins of intestines and liver and produce eggs that are eliminated within feces. If the eggs reach water, a ciliated larva is released and enters snails to develop into a larva infective to man and other vertebrates. Most infections evolve without overt disease, but severe intestinal, hepatic, pulmonary and cerebro-medulary dysfunctions may occur after many years. Definitive diagnosis is made through the identification of eggs in stool. Classical diagnostic methods fail to detect infection when the number of eggs is low (e.g., in areas where control measures have decreased the intensity of infection or in the case of light infections in travelers who have had only brief exposure). A new and very sensitive method is reported here, in which eggs are isolated from large amounts of feces through their interaction with magnetic beads. After incubation with the fecal sediment, eggs co-migrate with the beads towards a magnet attached to the test tube. This improvement in diagnostic methodology will strengthen efforts to control schistosomiasis.