Peer-reviewed veterinary case report
A highly sensitive DNA bead-based suspension array for the detection and species identification of bovine piroplasms.
- Journal:
- International journal for parasitology
- Year:
- 2012
- Authors:
- Ros-García, Amaia et al.
- Affiliation:
- Department of Animal Health · Spain
Plain-English summary
Researchers have developed a new testing method to quickly and accurately identify harmful parasites called piroplasms in cattle, which are spread by ticks. The new test, using a system called Luminex, is faster and more reliable than the older method known as Reverse Line Blot, which was complicated and could lead to mistakes in reading results. This new approach can detect very low levels of these parasites in the blood, making it easier to find infected animals. By testing 214 blood samples, the researchers confirmed that the Luminex method is both more sensitive and specific than the previous technique. Overall, this new test can help protect cattle from these parasites and improve efforts to control their spread.
Abstract
Piroplasms are among the most harmful tick-borne pathogens for livestock and sensitive and specific diagnostic methods for rapid detection and identification of the different species are needed for effective control. Reverse Line Blot has been the molecular technique of choice but it is laborious, time-consuming and highly susceptible to subjective variation in the interpretation of the hybridisation signal. Here, an oligonucleotide multiplex suspension microarray (Luminex® microsphere system) was developed for bovine piroplasms. Probes previously used in Reverse Line Blot for Babesia divergens, Babesia bovis, Babesia occultans, Babesia bigemina and Theileria buffeli, and a catch-all Theileria and Babesia control probe, were included in the Luminex assay together with newly designed probes for Theileria annulata and Babesia major. An internal amplification control that was detected with a Luminex probe was included to monitor for inhibition. Serially diluted linearised recombinant plasmids of the different species were used to assess the analytical sensitivity and specificity, and the detection limit of the Luminex assay was determined using serial dilutions of infected blood from an animal with a known level of T. annulata parasitaemia. The assay was then validated on 214 bovine blood samples analysed in parallel by Reverse Line Blot and Luminex. The Luminex assay proved to be highly specific and more sensitive than Reverse Line Blot, detecting 0.05 parasites/μl of blood. Technically, the Luminex procedure was rapid, provided high throughput screening, transformed the subjective interpretation of Reverse Line Blot results into numerical objective values, and allowed more flexibility in array preparation than Reverse Line Blot. The method described herein can substantially improve the detection of piroplasm carriers and thus better protect livestock trade and facilitate preventive control programs.
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Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/22233830/