A Real-Time PCR based assay for determining parasite to host ratio and parasitaemia in the clinical samples of Bovine Theileriosis
Open Access
- 18 October 2018
- journal article
- research article
- Published by Springer Science and Business Media LLC in Scientific Reports
- Vol. 8 (1), 1-7
- https://doi.org/10.1038/s41598-018-33721-3
Abstract
Theileria annulata is an intracellular parasite that causes active and latent forms of bovine theileriosis. Diagnosis of the disease is primarily based on traditional methods such as microscopy, however, PCR based methods have proven to be superior in the absence of clear disease symptoms. However, diagnosis is difficult in cases of lower parasitaemia by conventional PCR. Hence, a rapid and sensitive method which can detect early infection and low parasite load is required. Therefore, we have developed an absolute quantification based real-time PCR (qPCR) assay. Reference standard curve using recombinant plasmids of a host (hprt) and a parasite gene (tasp) was constructed, and the assay was initially standardised using in vitro T. annulata cell lines. Further, 414 blood samples from suspected theileriosis cases were also evaluated using qPCR. The assay can estimate host to parasite ratios, calculate parasitaemia and treatment effectiveness in the clinical cases of theileriosis. In comparison with the conventional PCR results, 44 additional positive cases were found. Therefore, the assay holds importance in a clinical setting due to its ability to quantify the parasite load in clinical samples. It may be further used in distinguishing active and latent theileriosis infections and detection of drug resistance in the field.Keywords
This publication has 35 references indexed in Scilit:
- Evaluation of a Real-Time PCR Test for the Detection and Discrimination of Theileria Species in the African Buffalo (Syncerus caffer)PLOS ONE, 2013
- Revisiting the Tams1-encoding gene as a species-specific target for the molecular detection of Theileria annulata in bovine blood samplesTicks and Tick-borne Diseases, 2013
- Development and evaluation of a real-time PCR assay for the quantitative detection of Theileria annulata in cattleParasites & Vectors, 2012
- Evaluation of cytochrome b as a sensitive target for PCR based detection of T. annulata carrier animalsVeterinary Parasitology, 2010
- In vivo evidence for the resistance of Theileria annulata to buparvaquoneVeterinary Parasitology, 2010
- Bovine immunity – a driver for diversity in Theileria parasites?Trends in Parasitology, 2009
- Development and evaluation of a real-time polymerase chain reaction test for the detection of Theileria parva infections in Cape buffalo (Syncerus caffer) and cattleVeterinary Parasitology, 2008
- From molecule to diagnostic tool: Theileria annulata surface protein TaSPZeitschrift für Parasitenkunde, 2007
- Genome of the Host-Cell Transforming Parasite Theileria annulata Compared with T. parvaScience, 2005
- Treatment of experimentally induced Theileria annulata infection in cross-bred calves with buparvaquoneVeterinary Parasitology, 1988