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Home / Equine Research Bank / Trop Anim Health Prod / Comparison of polymerase chain reaction methods for the dete...
Tropical animal health and production2007; 39(5); 369-374; doi: 10.1007/s11250-007-9025-1

Comparison of polymerase chain reaction methods for the detection of Theileria equi infection using whole blood compared with pre-extracted DNA samples as PCR templates.

Abstract: Rapid, efficient, and reproducible procedures for isolating DNA before PCR gene amplification are essential for the diagnosis of piroplasms. In this study, we evaluated the ease and reliability of detecting Theileria equi by PCR using pre-extracted DNA samples (by QIAamp DNA Mini Kit and phenol-chloroform methods) compared with blood spotted on FTA cards as PCR templates. Although minimal variations in limit of detection were observed among the methods compared, overall, the use of pre-extracted DNA samples and blood spotted on FTA cards had comparable detection limits. These results indicate that T. equi infection can be efficiently detected directly from FTA cards by PCR without the need for pre-extraction of DNA from blood samples.
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Publication Date: 2007-10-20 PubMed ID: 17944307DOI: 10.1007/s11250-007-9025-1Google Scholar: Lookup
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  • Comparative Study
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

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.

The study investigates the effectiveness of using different methods to detect the parasite Theileria equi. It found that the same detection limits were possible whether pre-extracted DNA samples were used, or if blood spotted directly on FTA cards without pre-extraction was used.

Research Objective and Methodology

  • The primary objective of the research was to evaluate the ease and reliability of detecting Theileria equi, a piroplasm that causes disease in horses, through polymerase chain reaction (PCR) methods.
  • The testing was done using pre-extracted DNA samples and comparing it with blood samples directly spotted on Flinders Technology Associates (FTA) cards.
  • The pre-extraction of DNA samples was performed using two methods – the QIAamp DNA Mini Kit and phenol-chloroform methods.

Results and Observations

  • Only minimal variations were observed in limit of detection among the methods tested. The limit of detection refers to the smallest concentration or absolute amount of analyte (the substance being measured) that can be reliably detected. A lower limit of detection is often more desirable in many kinds of testing.
  • Both using pre-extracted DNA samples and putting blood directly on FTA cards as PCR templates displayed comparable detection limits.
  • This equivalency suggests that the extra step of pre-extracting DNA might not be necessary and could simplify the detection process.

Significance and Implications

  • The results imply that Theileria equi infection can be efficiently detected directly from FTA cards by PCR, eliminating the need for the pre-extraction of DNA from blood samples.
  • The outcome of the study could potentially contribute to making DNA isolation procedures quicker, more cost-effective, and easier to perform, especially in field settings or areas with limited resources or equipment.
  • However, further validation is required with larger sample sizes and diverse field samples before this method can be consistently applied in practice.

Cite This Article

APA
Alhassan A, Iseki H, Kim C, Yokoyama N, Igarashi I. (2007). Comparison of polymerase chain reaction methods for the detection of Theileria equi infection using whole blood compared with pre-extracted DNA samples as PCR templates. Trop Anim Health Prod, 39(5), 369-374. https://doi.org/10.1007/s11250-007-9025-1

Publication

Tropical animal health and production
ISSN: 0049-4747
NlmUniqueID: 1277355
Country: United States
Language: English
Volume: 39
Issue: 5
Pages: 369-374

Researcher Affiliations

Alhassan, A
  • National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.
Iseki, H
    Kim, C
      Yokoyama, N
        Igarashi, I

          MeSH Terms

          • Animals
          • DNA, Protozoan / chemistry
          • DNA, Protozoan / genetics
          • Ghana
          • Horse Diseases / blood
          • Horse Diseases / diagnosis
          • Horse Diseases / parasitology
          • Horses
          • Polymerase Chain Reaction / methods
          • Polymerase Chain Reaction / veterinary
          • Protozoan Proteins / chemistry
          • Protozoan Proteins / genetics
          • RNA, Ribosomal, 18S / chemistry
          • RNA, Ribosomal, 18S / genetics
          • Theileria / genetics
          • Theileria / isolation & purification
          • Theileriasis / blood
          • Theileriasis / parasitology

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          Citations

          This article has been cited 5 times.
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