Angiostrongyliasis is a disease caused by Angiostrongylus nematodes that is present worldwide. The infections with the highest impact on human and animal health are caused by A. cantonensis, A. costaricensis, and A. vasorum. Clinical forms of the disease in humans are eosinophilic meningitis and abdominal angiostrongyliasis, while the most common effect on dogs are cardiopulmonary damages. It is deemed as an emerging disease as the result of the global dissemination of the African snail Lissachatina fulica, an intermediary host of these parasites. The few diagnostic methods for Angiostrongylus spp. are for a single parasite, costly, low sensitive and not available worldwide . It is urgent to develop a sensitive, specific and accessible diagnostic tool for the control of human and animal angiostrongyliasis. Objective: Standardization of a multiplex qPCR for the diagnosis of three Angiostrongylus of clinical importance. Results: The design of primer and taq-man probes allowed the identification of the parasites from the ITS-2 sequence shared by the three species. qPCR did not amplify African snail DNA, human DNA and other parasites. The threshold cycle values for positive DNA controls were: 21 for Angiostrongylus cantonensis, 22 for A. costaricensis, and 31 for A. vasorum. In negative controls, the threshold cycle was zero. qPCR showed an amplification efficiency of 2 (100%). Conclusions: The standardized technique was able to identify and differentiate specifically the three Angiostrongylus species in the same qPCR reaction.
Dr. Ruben Eduardo Varela Miranda, is PhD from Salamanca University (Spain). He is proffesor in the school medicine and science basic in the Universidad Santiago de Cali (Colombia). He has published papers in reputed journals and his has a line the research in drug discovery trought identification new molecular target in proteins kinase in trypanosomatid parasites. The professor in 2018 developed a molecular test for the identification of three parasites Angiostrongylus (A. cantonensis, A.vasorum, A. costaricensis) by multiple PCR in African snails.