title: Comparative evaluation of the sensitivity of LAMP, PCR and in vitro culture methods for the diagnosis of equine piroplasmosis.
abstract: The sensitivity of LAMP, PCR and in vitro culture methods for the detection of Theileria equi and Babesia caballi was evaluated using tenfold serially diluted culture parasites. On day 1 post-culture, both T. equi and B. caballi parasites could only be observed at 1% parasite dilution from the in vitro culture method, whereas LAMP could detect up to 1 x 10(-3)% of both T. equi and B. caballi parasite dilutions, whilst PCR could detect 1 x 10(-3)% T. equi and 1 x 10(-1)% B. caballi parasite dilutions. On day 7 post-culture, the detection limit for T. equi and B. caballi in the in vitro culture increased up to 1 x 10(-6)%, whereas LAMP detection limit increased to 1 x 10(-10)% for both parasites, whilst the PCR detection limit increased to 1 x 10(-10)% and 1 x 10(-6)% for T. equi and B. caballi, respectively. Furthermore, LAMP and PCR amplified the T. equi DNA extracted from the organs of an experimentally infected horse. This study further validates LAMP as an alternative molecular diagnostic tool, which can be used in the diagnosis of early infections of equine piroplasmosis and together with PCR can also be used as supplementary methods during post-mortems.
authors.affiliation: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.
authors.forename: Andy
authors.initials: A
authors.lastname: Alhassan
authors.forename: Yadav
authors.initials: Y
authors.lastname: Govind
authors.forename: Nguyen Thanh
authors.initials: NT
authors.lastname: Tam
authors.forename: Oriel M M
authors.initials: OM
authors.lastname: Thekisoe
authors.forename: Naoaki
authors.initials: N
authors.lastname: Yokoyama
authors.forename: Noboru
authors.initials: N
authors.lastname: Inoue
authors.forename: Ikuo
authors.initials: I
authors.lastname: Igarashi
mesh_terms: D000818:Animals; D001403:Babesia / Q000302:isolation & purification*; D001404:Babesiosis / Q000175:diagnosis / Q000662:veterinary*; D046508:Culture Techniques; D006734:Horse Diseases / Q000175:diagnosis*; D006736:Horses / Q000469:parasitology; D021141:Nucleic Acid Amplification Techniques; D012680:Sensitivity and Specificity; D016794:Theileria / Q000302:isolation & purification*