Journal
VETERINARY PARASITOLOGY
Volume 165, Issue 1-2, Pages 36-40Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.vetpar.2009.06.036
Keywords
Babesia orientalis; LAMP; 18S rRNA; Babesiosis
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Funding
- National Natural Science Foundation of China (NSFC) [30671575]
- Ministry of Education, PR China [NCET06-0668]
- South African National Research Foundation (NRF) [67147]
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Loop-mediated isothermal amplification (LAMP) is a rapid method with high specificity and efficiency under isothermal condition using a set of four specifically designed primers that recognize six distinct sequences on the target gene. In this study, a LAMP method was developed for specific detection of Babesia orientalis in water buffalo (Bubalus babalis, Linnaeus, 1758). Four primers were designed from the V4 hypervariable region of the 18S rRNA gene of B. orientalis. Blood samples were collected from B. orientalis experimentally infected water buffalo as well as from 165 water buffalo from eight different regions of the Hubei province, south China. Genomic DNA was extracted, subjected to the LAMP assay and compared with results obtained using a previously described semi-nested PCR. The LAMP assay proofed to be B. orientalis specific and more sensitive than the semi-nested PCR. While previously B. orientalis had not been reported north of the Yangtse River, our results show that B. orientalis has spread to the north of the river. This could pose a serious threat to the water buffalo industry. (C) 2009 Elsevier B.V. All rights reserved.
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