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Home / Equine Research Bank / Parasitol Res / Comparison of PCR-based methods for the detection of Babesia...
Parasitology research2021; 120(6); 2157-2164; doi: 10.1007/s00436-021-07153-4

Comparison of PCR-based methods for the detection of Babesia caballi and Theileria equi in field samples collected in Central Italy.

Abstract: Equine piroplasmosis (EP) is a disease of equids caused by Theileria equi and Babesia caballi, members of the order Piroplasmida, transmitted by several species of ticks. As the disease is endemic in many countries, a clinical examination or a serological test are required prior to movement of horses to prove freedom from infection and to avoid the introduction of EP with its sanitary and economic impact, especially in areas where it is absent. Currently, numerous diagnostic PCR protocols are available, some of which are recommended by the World Organisation for Animal Health (OIE). In order to adopt this diagnostic method, the Italian National Reference Centre for Equine Diseases (NRC-ED) conducted a preliminary comparison between an end-point PCR, nested PCR, real-time PCR, and commercial real-time PCR, for the detection of T. equi and B. caballi, respectively. One hundred and three field samples, collected during spring-summer 2013 in Latium and Tuscany regions, were employed for the study, and results discordant between detection assays were confirmed by sequencing. The reference assay was defined as that showing the highest sensitivity, and the relative sensitivity (rSe) and specificity (rSp) of the other methods were estimated referring to this assay. Agreement between methods was estimated by calculating the concordance between each pair of methods. Although no statistical differences were detected among PCR-based methods, the non-commercial real-time PCR assays seemed to be the most suitable for detection of T. equi and B. caballi, respectively. An important advantage of direct PCR detection of the pathogen, in comparison to indirect detection using serological methods, is that it allows specific treatment against the causative pathogen species responsible of the infection as well as for the definition of the infectious status of an animal for international movement.
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Publication Date: 2021-04-15 PubMed ID: 33855619PubMed Central: 3505731DOI: 10.1007/s00436-021-07153-4Google Scholar: Lookup
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  • Comparative Study
  • Journal Article
  • Validation Study

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research evaluates the comparative efficacy of different PCR-based diagnostic methods for detecting two pathogens causing equine piroplasmosis, a disease which impacts horses and is transmitted by ticks.

Objective of the Study

  • The main goal of the study was to compare the effectiveness of four PCR-based diagnostic methods: end-point PCR, nested PCR, real-time PCR, and commercial real-time PCR. These methods were used to detect two pathogens, Theileria equi and Babesia caballi, which are responsible for equine piroplasmosis, a disease prevalent in horses and other equids.

Disease background and Significance

  • Equine piroplasmosis is a disease of horses transmitted by several species of ticks. The disease is endemic in many countries and requires meticulous diagnostic tests to prove a horse is infection-free before it can be moved to a new location.
  • Accurate detection of the disease-causing pathogens is important to avoid the sanitary and economic impacts of the disease, especially in areas where it is not currently present.

Methodology

  • The study used 103 field samples collected from two regions in Italy during the spring-summer season of 2013.
  • The diagnostic methods compared were end-point PCR, nested PCR, real-time PCR, and commercial real-time PCR.
  • Results that were inconsistent between detection assays were confirmed by sequencing.

Findings

  • The research determined that although there were no statistical differences among the PCR-based methods, non-commercial real-time PCR tests were most suitable for detecting T. equi and B. caballi.
  • The reference assay (the assay with the highest sensitivity) was used to estimate the relative sensitivity and specificity of the other diagnostic methods. The concordance between each pair of methods was also calculated.

Significance of PCR-based Detection

  • PCR-based pathogen detection has an important advantage over indirect detection using serological methods: it allows for specific treatment against the causative species responsible for the infection.
  • Furthermore, it provides definite information on the infectious status of the animal, which is crucial for international movement of horses.

Cite This Article

APA
(2021). Comparison of PCR-based methods for the detection of Babesia caballi and Theileria equi in field samples collected in Central Italy. Parasitol Res, 120(6), 2157-2164. https://doi.org/10.1007/s00436-021-07153-4

Publication

Parasitology research
ISSN: 1432-1955
NlmUniqueID: 8703571
Country: Germany
Language: English
Volume: 120
Issue: 6
Pages: 2157-2164

Researcher Affiliations

MeSH Terms

  • Animals
  • Babesia / genetics
  • Babesia / isolation & purification
  • Babesiosis / epidemiology
  • Babesiosis / parasitology
  • Horse Diseases / epidemiology
  • Horse Diseases / parasitology
  • Horses
  • Italy / epidemiology
  • Molecular Diagnostic Techniques / veterinary
  • Polymerase Chain Reaction / methods
  • Polymerase Chain Reaction / veterinary
  • Real-Time Polymerase Chain Reaction / veterinary
  • Retrospective Studies
  • Theileria / genetics
  • Theileria / isolation & purification
  • Theileriasis / epidemiology
  • Theileriasis / parasitology

Grant Funding

  • IZSLT 16/11 / Ministero della Salute

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Citations

This article has been cited 4 times.
  1. Yao XY, Yu SQ, Tian N, Wang F, Li SZ, Li LH. Nucleic Acid Prevalence of Zoonotic Babesia in Humans, Animals and Questing Ticks, a Systematic Review and Meta-Analysis. Trop Med Infect Dis 2023 Feb 22;8(3).
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  2. Lv K, Zhang Y, Yang Y, Liu Z, Deng L. Development of Nested PCR and Duplex Real-Time Fluorescence Quantitative PCR Assay for the Simultaneous Detection of Theileria equi and Babesia caballi. Front Vet Sci 2022;9:873190.
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