Semi-nested PCR detection of Babesia orientalis in its natural hosts Rhipicephalus haemaphysaloides and buffalo

Loop-mediated isothermal amplification (LAMP) detection of Babesia orientalis in water buffalo (Bubalus babalis, Linnaeus, 1758) in China

“…It was first reported in 1984 in China (Chen, 1984) to occur in nine provinces (South of China) with a high prevalence in endemic areas, and it was described as B. orientalis in 1997 (Liu et al, 1997). A semi-nested PCR assay targeting the 18S rRNA gene was developed for the detection of B. orientalis in water buffalo and the potential tick vector R. haemaphysaloides (Liu et al, 2007). In this study, we describe the development of a simple, sensitive and cost-efficient loop-mediated isothermal amplification (LAMP) assay for the specific detection of B. orientalis in water buffalo samples.…”

“…In this study, we describe the development of a simple, sensitive and cost-efficient loop-mediated isothermal amplification (LAMP) assay for the specific detection of B. orientalis in water buffalo samples. Results were compared with that obtained using the semi-nested PCR of Liu et al (2007).…”

“…Two 1-year-old water buffalo, free of B. orientalis infection as confirmed with semi-nested PCR (Liu et al, 2007), were splenectomised 14 days prior to B. orientalis infection. The B. orientalis Wuhan strain, previously isolated from the Hubei province, China and preserved in liquid nitrogen (Liu et al, 2005), were used to infect the buffalo.…”

“…The primers of semi-nested PCR were designed based on the sequence of the 18S rRNA gene for B. orientalis (Liu et al, 2007). PCR products were visualized using 2% ethidium bromide-stained agarose gel electrophoresis.…”

“…et al, 1993;Smeenk et al, 2000;Almeria et al, 2001;Oliveira-Sequeira et al, 2005;Liu et al, 2007;Martins et al, 2008;Sibeko et al, 2008). These techniques, however, rely on the availability of expensive equipment such as thermocyclers.…”