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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
immunology and microbiology
Sensitive and specific detection of Cryptosporidium species in PCR-negative samples by loop-mediated isothermal DNA amplification and confirmation of generated LAMP products by sequencing
Veterinary Parasitology, Volume 158, No. 1-2, Year 2008
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Description
Three LAMP (loop-mediated isothermal DNA amplification) assays were applied to detect Cryptosporidium species DNA in a total number of 270 fecal samples originating from cattle, sheep and horses in South Africa. DNA was extracted from 0.5 g of fecal material. Results of LAMP detection were compared to those obtained by nested PCR targeting the Cryptosporidium 18 small subunit rRNA (18S) gene. All samples were negative by nested PCR, while up to one-third of samples were positive by LAMP assays. The SAM-1 LAMP assay, shown to detect C. parvum, C. hominis and C. meleagridis, amplified Cryptosporidium DNA in 36 of 107 cattle (33.64%), in 26 of 85 sheep (30.5%) and in 17 of 78 horses (21.79%). The HSP LAMP specific to C. muris and C. andersoni, amplified Cryptosporidium DNA in one cow (0.9%), five sheep (5.8%) and seven horses (8.9%). The gp60 LAMP assay, shown to detect C. parvum produced no amplified Cryptosporidium DNA, likely due to low sample DNA concentrations. The specificity of LAMP assays was confirmed by sequencing of the LAMP products generated in positive samples. Sequence products from the three LAMP assays showed high identity to the target gene sequences confirming the specificity of LAMP. In this study, the LAMP procedure was clearly superior to nested PCR in the detection of Cryptosporidium species DNA. Use of LAMP is proposed as an efficient and effective tool for epidemiologic survey studies including screening of healthy animals in which Cryptosporidium oocyst shedding is characteristically low and likely below the detection limit of PCR in conventional sample concentrates. © 2008 Elsevier B.V. All rights reserved.
Authors & Co-Authors
Bakheit, Mohammed Ahmed
Japan, Obihiro
Obihiro University of Agriculture and Veterinary Medicine
Torra, Dena
Japan, Obihiro
Obihiro University of Agriculture and Veterinary Medicine
Palomino, Lily A.
Japan, Obihiro
Obihiro University of Agriculture and Veterinary Medicine
Thekisoe, Oriel M.M.
Japan, Obihiro
Obihiro University of Agriculture and Veterinary Medicine
Mbati, Peter Amunga
South Africa, Phuthaditjhaba
Parasitology Research Program
South Africa, Thohoyandou
University of Venda
Ongerth, Jerry E.
Australia, Wollongong
University of Wollongong
Karanis, Panagiotis
Japan, Obihiro
Obihiro University of Agriculture and Veterinary Medicine
Germany, Koln
Universität zu Köln
Statistics
Citations: 113
Authors: 7
Affiliations: 5
Identifiers
Doi:
10.1016/j.vetpar.2008.09.012
ISSN:
03044017
Research Areas
Genetics And Genomics
Study Design
Cross Sectional Study
Study Approach
Quantitative
Study Locations
South Africa