Next Article in Journal / Special Issue
Anaplasma phagocytophilum-Occupied Vacuole Interactions with the Host Cell Cytoskeleton
Previous Article in Journal
Comparative Pathogenesis of Cancers in Animals and Humans
Previous Article in Special Issue
Ehrlichioses: An Important One Health Opportunity
Article Menu

Export Article

Open AccessArticle
Vet. Sci. 2016, 3(3), 23; doi:10.3390/vetsci3030023

An Evaluation of Quantitative PCR Assays (TaqMan® and SYBR Green) for the Detection of Babesia bigemina and Babesia bovis, and a Novel Fluorescent-ITS1-PCR Capillary Electrophoresis Method for Genotyping B. bovis Isolates

1
Department of Agriculture & Fisheries, Agri-Science Queensland, Animal Science, Dutton Park QLD 4102, Australia
2
Department of Agriculture & Fisheries, Biosecurity Queensland, Tick Fever Centre, Wacol QLD 4076, Australia
3
Queensland Alliance for Agriculture & Food Innovation, University of Queensland, Centre for Animal Science, St Lucia QLD 4072, Australia
*
Authors to whom correspondence should be addressed.
Academic Editor: Ulrike Munderloh
Received: 10 June 2016 / Revised: 30 August 2016 / Accepted: 7 September 2016 / Published: 13 September 2016
(This article belongs to the Special Issue Comparative Studies in Tick-Borne Diseases in Animals and Humans)
View Full-Text   |   Download PDF [1413 KB, uploaded 13 September 2016]   |  

Abstract

Babesia spp. are tick-transmitted haemoparasites causing tick fever in cattle. In Australia, economic losses to the cattle industry from tick fever are estimated at AUD$26 Million per annum. If animals recover from these infections, they become immune carriers. Here we describe a novel multiplex TaqMan qPCR targeting cytochrome b genes for the identification of Babesia spp. The assay shows high sensitivity, specificity and reproducibility, and allows quantification of parasite DNA from Babesia bovis and B. bigemina compared to standard PCR assays. A previously published cytochrome b SYBR Green qPCR was also tested in this study, showing slightly higher sensitivity than the Taqman qPCRs but requires melting curve analysis post-PCR to confirm specificity. The SYBR Green assays were further evaluated using both diagnostic submissions and vaccinated cattle (at 7, 9, 11 and 14 days post-inoculation) showed that B. bigemina can be detected more frequently than B. bovis. Due to fewer circulating parasites, B. bovis detection in carrier animals requires higher DNA input. Preliminary data for a novel fluorescent PCR genotyping based on the Internal Transcribed Spacer 1 region to detect vaccine and field alleles of B. bovis are described. This assay is capable of detecting vaccine and novel field isolate alleles in a single sample. View Full-Text
Keywords: babesiosis; carrier; diagnosis; qPCR; cytochrome b; genotyping; ITS1; vaccines babesiosis; carrier; diagnosis; qPCR; cytochrome b; genotyping; ITS1; vaccines
Figures

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

Supplementary material

Scifeed alert for new publications

Never miss any articles matching your research from any publisher
  • Get alerts for new papers matching your research
  • Find out the new papers from selected authors
  • Updated daily for 49'000+ journals and 6000+ publishers
  • Define your Scifeed now

SciFeed Share & Cite This Article

MDPI and ACS Style

Zhang, B.; Sambono, J.L.; Morgan, J.A.T.; Venus, B.; Rolls, P.; Lew-Tabor, A.E. An Evaluation of Quantitative PCR Assays (TaqMan® and SYBR Green) for the Detection of Babesia bigemina and Babesia bovis, and a Novel Fluorescent-ITS1-PCR Capillary Electrophoresis Method for Genotyping B. bovis Isolates. Vet. Sci. 2016, 3, 23.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Vet. Sci. EISSN 2306-7381 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top