caa a22 4500 445339748 CHVBK 20180317142808.0 cr unu---uuuuu 170323e20111201xx s 000 0 eng 10.1007/s00436-011-2414-4 doi (NATIONALLICENCE)springer-10.1007/s00436-011-2414-4 Improved detection of canine Angiostrongylus vasorum infection using real-time PCR and indirect ELISA [Elektronische Daten] [Ryan Jefferies, Eric Morgan, Jenny Helm, Matthew Robinson, Susan Shaw] This study reports the development of a real-time PCR assay and an indirect ELISA to improve on current detection of canine Angiostrongylus vasorum infection. A highly specific fluorescent probe-based, real-time PCR assay was developed to target the A. vasorum second internal transcribed spacer region and detected DNA in EDTA blood, lung tissue, broncho-alveolar larvage fluid, endotracheal mucus, pharyngeal swabs and faecal samples. PCR was fast (∼1h), highly efficient when using EDTA blood samples, consistently detected a single molecule of parasite DNA and did not amplify DNA from other parasitic nematodes or definitive host species. An indirect ELISA was also developed using the soluble protein fraction from adult A. vasorum worms. Some cross-reactive antigen recognition was observed when tested against sera from dogs infected with Crenosoma vulpis (n = 8), Toxocara canis (n = 5) and Dirofilaria immitis (n = 5). This was largely overcome by setting the cut-off for a positive result at an appropriately high level. Field evaluation of the real-time PCR and ELISA was conducted by testing sera and EDTA blood from dogs with suspected A. vasorum infection (n = 148) and compared with the Baermann's larval migration test in faeces. Thirty-one dogs were positive by at least one test. Of these, 20 (65%) were detected by the Baermann method, 18 (58%) by blood PCR, 24 (77%) by ELISA and 28 (90%) by blood PCR and ELISA together. Combined testing using real-time PCR and ELISA therefore improved the detection rate of A. vasorum infection and holds promise for improved clinical diagnosis and epidemiological investigation. Springer-Verlag, 2011 Jefferies Ryan School of Clinical Veterinary Sciences, University of Bristol, Langford, BS40 5DU, Bristol, UK aut Morgan Eric School of Biological Sciences, University of Bristol, BS8 1UG, Bristol, UK aut Helm Jenny School of Veterinary Medicine, College of Medical, Veterinary & Life Sciences, University of Glasgow, Bearsden Road, Bearsden, G61 1QH, Glasgow, UK aut Robinson Matthew School of Clinical Veterinary Sciences, University of Bristol, Langford, BS40 5DU, Bristol, UK aut Shaw Susan School of Clinical Veterinary Sciences, University of Bristol, Langford, BS40 5DU, Bristol, UK aut Parasitology Research Springer-Verlag 109/6(2011-12-01), 1577-1583 0932-0113 109:6<1577 2011 109 436 https://doi.org/10.1007/s00436-011-2414-4 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00436-011-2414-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Jefferies Ryan School of Clinical Veterinary Sciences, University of Bristol, Langford, BS40 5DU, Bristol, UK aut NATIONALLICENCE 700 1- Morgan Eric School of Biological Sciences, University of Bristol, BS8 1UG, Bristol, UK aut NATIONALLICENCE 700 1- Helm Jenny School of Veterinary Medicine, College of Medical, Veterinary & Life Sciences, University of Glasgow, Bearsden Road, Bearsden, G61 1QH, Glasgow, UK aut NATIONALLICENCE 700 1- Robinson Matthew School of Clinical Veterinary Sciences, University of Bristol, Langford, BS40 5DU, Bristol, UK aut NATIONALLICENCE 700 1- Shaw Susan School of Clinical Veterinary Sciences, University of Bristol, Langford, BS40 5DU, Bristol, UK aut NATIONALLICENCE 773 0- Parasitology Research Springer-Verlag 109/6(2011-12-01), 1577-1583 0932-0113 109:6<1577 2011 109 436 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer