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Loop-mediated isothermal amplification: rapid detection of Angiostrongylus cantonensis infection in Pomacea canaliculata

DOI: 10.1186/1756-3305-4-204

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Abstract:

We used a loop-mediated isothermal amplification (LAMP) assay, which was performed using Bst DNA polymerase. Reactions amplified the A. cantonensis 18S rRNA gene and demonstrated high sensitivity; as little as 1 fg of DNA was detected in the samples. Furthermore, no cross-reactivity was found with other parasites such as Toxoplasma gondii, Plasmodium falciparum, Schistosoma japonicum, Clonorchis sinensis, Paragonimus westermani and Anisakis. Pomacea canaliculata snails were exposed to A. cantonensis first-stage larvae (L1) in the laboratory, and L3 were observed in the snails thirty-five days after infection. All nine samples were positive as determined by the LAMP assay for A. cantonensis, which was identified as positive by using PCR and microscopy, this demonstrates that LAMP is sensitive and effective for diagnosis.LAMP is an appropriate diagnostic method for the routine identification of A. cantonensis within its intermediate host snail P. canaliculata because of its simplicity, sensitivity, and specificity. It holds great promise as a useful monitoring tool for A. cantonensis in endemic regions.Angiostrongylus cantonensis can be found in the lungs and arteries of insectivores, rodents, canines and felines [1-6]. They are prevalent in the Pacific islands and Southeast Asia, and are the most common cause of eosinophilic meningitis in humans in areas where the parasite is endemic [7]. The definitive hosts of A. cantonensis are various species of rats. Modes of transmission of this parasite include ingestion of raw or undercooked snails and fresh leafy vegetables contaminated by infective third-stage larvae (L3) [8]. First-stage larvae (L1) of A. cantonensis grow to infective L3 in intermediate host snails. This disease is difficult to detect because of the long incubation period in patients and few diagnostic symptoms. Thus, in order to control A. cantonensis, efforts should be directed towards building a surveillance system for the intermediate host snails of th

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