Severe Delayed Hemolysis Associated with Regulated Parenteral Antimalarial Drug

Techapp1.pdf) were designed to amplify short sequences of mitochondrial DNA. On the basis of primer specificity and amplicon size, we determined host and parasite species (online Technical Appendix Figure 1). DNA extraction and PCR were performed in a laboratory dedicated to environmental samples (for a complete description of methods, see online Technical Appendix). DNA was successfully extracted and amplified from 119 (86.9%) of 137 fecal samples. Of usable samples, 28 (23.5%) were from red foxes and 91 (76.5%) were from dogs. Two fox fecal samples (7.1%; 95% binomial CIs 0.9%–23.5%) were infected with E. multilocularis tape-worms; none of the dog samples were infected. To verify parasite identification, we amplified DNA from the 2 E. multilocularis–positive samples with E. mul-tilocularis–specific primers and sequenced the amplification products. To verify host species identification, we used primers that produced longer amplification products (327 and 197 bp) than the corresponding PCR primers and sequenced amplification products from 5 fox samples and 5 dog samples. Sequencing procedures were performed according to the methods of Saarma et al. (10). Sequences from both E. multilocularis–positive samples showed 100% identity with an E. multilocularis tape-worm sequence (GenBank accession no. AB018440) (on-line Technical Appendix Figure 2). All sequenced fox and dog samples also belonged to the corresponding species. To estimate the sensitivity of this noninvasive genetic method, we determined the number of E. multilocularis eggs necessary to obtain a positive PCR result (online Technical Appendix Figure 3). One egg was sufficient to give an E. multilocularis tapeworm–specific result. In summary, we developed a noninvasive genetic method that identifies E. multilocularis tapeworms and their host species in carnivore fecal samples found in urban environments. Furthermore, these tapeworms can even be detected in fecal samples from red foxes when only 1 parasite egg is present. Thus, this method is highly sensitive and discriminatory and can be used with degraded fecal samples to monitor E. multilocularis tapeworms and their hosts. et al. A real-time multiplex-nested PCR system for coprological diagnosis of Echinococcus multilocularis and host species. Echinococcus granulosus G8 in Eurasia and a reappraisal of the phylogenetic relationships of " genotypes " G5–G10. Parasitology. A, et al. A novel phylogeny for the genus Echinococcus, based on nuclear data, challenges relationships based on mitochondrial evidence .