A PCR method targeting internal transcribed spacers: the simultaneous detection of Babesia bigemina and Babesia bovis in cattle

Abstract

In this study, two pairs of oligonucleotide primers were designed according to the nucleotide sequence of the internal transcribed spacers (ITSs) of Babesia bigemina and B. bovis isolates from China. The primers were used in a multiplex PCR to detect parasite DNA in blood samples from cattle. There was no cross reactions with B. ovata, B. major, B. sp. Kashi, Theileria annulata, T. sergenti, T. sinensis or normal bovine DNA. The sensitivity of multiplex PCR assay was 1 pg and 10 pg DNA for B. bigemina and B. bovis, respectively. A total of 260 field blood samples collected from cattle in five provinces of China were analyzed by multiplex PCR and light microscopy. PCR testing revealed that 7.3% (19/260) and 5.8% (15/260) of cattle were positive for B. bigemina and B. bovis and 1.2% (3/260) of cattle were co-infected with B. bigemina and B. bovis. Using light microscopy, 2.3% (6/260) and 1.5% (4/260) of cattle were infected by B. bigemina and B. bovis, respectively, and no co-infection was found. The results showed that the multiplex PCR developed in the present study could be an alternative diagnostic tool for the detection of B. bovis and B. bigemina infection in cattle.

Epidemiological, Clinical and Diagnostic Studies on Blood Parasites in Cattle and Buffaloes in ElBehera Provence

“…Incidence of Babesia bigemina was 9.45%. This result nearly similar to the results obtained by Liu et al, (2014) who revealed that 7.3% (19/260) of cattle were positive for B. bigemina .This result disagree with the results obtained by Nayel et al, (2012) found that 54 (19%) were positive using direct smear and 62 (22.5%) were positive using PCR.…”

Identification of 12 Piroplasms Infecting Ten Tick Species in China Using Reverse Line Blot Hybridization

“…microplus were positive in terms of B. bigemina and B. bovis infections, respectively (Table IV). An earlier study in China reported a higher prevalence of B. bigemina and B. bovis (7.3% and 5.8%) by multiplex PCR assay, and 2.3% and 1.5% by light microscopy, respectively (Liu et al, 2014). Moreover, 1 R. sanguineus displayed a co-infection of T. annulata and B. bigemina, which is a new finding and suggests that the role of R. sanguineus in B. bigemina transmission needs to be experimentally validated.…”

“…Thus, the potential transmitted role of this tick for T. luwenshuni has to be further studied, and blood from these animals should be tested for piroplasm infection. The prevalence of B. bigemina and B. bovis reported was 0.6% and 0.2%, respectively, but high prevalence of B. bigemina and B. bovis in cattle was recently reported using PCR (Liu et al, 2014;Niu et al, 2015). Several studies showed that B. bigemina and B. bovis could be transmitted by the genus R.…”

The occurrence of pathogens in Rhipicephalus microplus ticks from cattle in Madagascar

“…PCR for Babesia bovis was carried out using the primers Bo5'-CTTGCGGCGATTTGGC-3' and BoR5'-CGTGAAGGAGCGGTGTAGAG-3', which can amplify a product of 408 bp from the nucleotide sequence of the internal transcribed spacers (ITSs) (Liu et al 2014). Amplification conditions were as follows: initial denaturation at 96 °C for 5 min, followed by 35 cycles of denaturation at 92 °C for 1 min, annealing at 56 °C for 1 min, extension at 72 °C for 1 min, and then a final extension at 72 °C for 1 min (Liu et al 2014).…”

“…Amplification conditions were as follows: initial denaturation at 96 °C for 5 min, followed by 35 cycles of denaturation at 92 °C for 1 min, annealing at 56 °C for 1 min, extension at 72 °C for 1 min, and then a final extension at 72 °C for 1 min (Liu et al 2014).…”