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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

          References

          This article includes 23 references
          1. Zhong KJ, Salas CJ, Shafer R, Gubanov A, Gasser RA Jr, Magill AJ, Forney JR, Kain KC. Comparison of IsoCode STIX and FTA Gene Guard collection matrices as whole-blood storage and processing devices for diagnosis of malaria by PCR.. J Clin Microbiol 2001 Mar;39(3):1195-6.
            pubmed: 11230458doi: 10.1128/JCM.39.3.1195-1196.2001google scholar: lookup
          2. Okamoto H, Takano E, Sugao T, Kage K, Okamoto E, Nishimura N, Ueda K. Direct amplification of Escherichia coli O157 vero toxin genes from human faeces by the polymerase chain reaction.. Ann Clin Biochem 1999 Sep;36 ( Pt 5):642-8.
            pubmed: 10505216doi: 10.1177/000456329903600513google scholar: lookup
          3. Böse R, Jorgensen WK, Dalgliesh RJ, Friedhoff KT, de Vos AJ. Current state and future trends in the diagnosis of babesiosis.. Vet Parasitol 1995 Mar;57(1-3):61-74.
            pubmed: 7597794doi: 10.1016/0304-4017(94)03111-9google scholar: lookup
          4. Alhassan A, Pumidonming W, Okamura M, Hirata H, Battsetseg B, Fujisaki K, Yokoyama N, Igarashi I. Development of a single-round and multiplex PCR method for the simultaneous detection of Babesia caballi and Babesia equi in horse blood.. Vet Parasitol 2005 Apr 20;129(1-2):43-9.
            pubmed: 15817201doi: 10.1016/j.vetpar.2004.12.018google scholar: lookup
          5. Rampersad J, Cesar E, Campbell MD, Samlal M, Ammons D. A field evaluation of PCR for the routine detection of Babesia equi in horses.. Vet Parasitol 2003 May 30;114(2):81-7.
            pubmed: 12781470doi: 10.1016/s0304-4017(03)00129-8google scholar: lookup
          6. Hänscheid T, Grobusch MP. How useful is PCR in the diagnosis of malaria?. Trends Parasitol 2002 Sep;18(9):395-8.
            pubmed: 12377256doi: 10.1016/s1471-4922(02)02348-6google scholar: lookup
          7. Bell-Sakyi L, Koney EB, Dogbey O, Walker AR. Incidence and prevalence of tick-borne haemoparasites in domestic ruminants in Ghana.. Vet Parasitol 2004 Sep 20;124(1-2):25-42.
            pubmed: 15350659doi: 10.1016/j.vetpar.2004.05.027google scholar: lookup
          8. Nicolaiewsky TB, Richter MF, Lunge VR, Cunha CW, Delagostin O, Ikuta N, Fonseca AS, da Silva SS, Ozaki LS. Detection of Babesia equi (Laveran, 1901) by nested polymerase chain reaction.. Vet Parasitol 2001 Oct 31;101(1):9-21.
            pubmed: 11587829doi: 10.1016/s0304-4017(01)00471-xgoogle scholar: lookup
          9. Birkenheuer AJ, Levy MG, Breitschwerdt EB. Development and evaluation of a seminested PCR for detection and differentiation of Babesia gibsoni (Asian genotype) and B. canis DNA in canine blood samples.. J Clin Microbiol 2003 Sep;41(9):4172-7.
            pubmed: 12958243doi: 10.1128/JCM.41.9.4172-4177.2003google scholar: lookup
          10. Hirata H, Ikadai H, Yokoyama N, Xuan X, Fujisaki K, Suzuki N, Mikami T, Igarashi I. Cloning of a truncated Babesia equi gene encoding an 82-kilodalton protein and its potential use in an enzyme-linked immunosorbent assay.. J Clin Microbiol 2002 Apr;40(4):1470-4.
            pubmed: 11923375doi: 10.1128/JCM.40.4.1470-1474.2002google scholar: lookup
          11. Boom R, Sol CJ, Salimans MM, Jansen CL, Wertheim-van Dillen PM, van der Noordaa J. Rapid and simple method for purification of nucleic acids.. J Clin Microbiol 1990 Mar;28(3):495-503.
            pubmed: 1691208doi: 10.1128/jcm.28.3.495-503.1990google scholar: lookup
          12. Mehlhorn H, Schein E. Redescription of Babesia equi Laveran, 1901 as Theileria equi Mehlhorn, Schein 1998.. Parasitol Res 1998 Jun;84(6):467-75.
            pubmed: 9660136doi: 10.1007/s004360050431google scholar: lookup
          13. Bork S, Yokoyama N, Matsuo T, Claveria FG, Fujisaki K, Igarashi I. Growth inhibitory effect of triclosan on equine and bovine Babesia parasites.. Am J Trop Med Hyg 2003 Mar;68(3):334-40.
            pubmed: 12685641
          14. Kappmeyer LS, Perryman LE, Knowles DP Jr. A Babesia equi gene encodes a surface protein with homology to Theileria species.. Mol Biochem Parasitol 1993 Nov;62(1):121-4.
            pubmed: 8114813doi: 10.1016/0166-6851(93)90185-zgoogle scholar: lookup
          15. Friedhoff KT, Soulé C. An account on equine babesioses.. Rev Sci Tech 1996 Sep;15(3):1191-201.
            pubmed: 9025154doi: 10.20506/rst.15.3.972google scholar: lookup
          16. Fahle GA, Fischer SH. Comparison of six commercial DNA extraction kits for recovery of cytomegalovirus DNA from spiked human specimens.. J Clin Microbiol 2000 Oct;38(10):3860-3.
            pubmed: 11015421doi: 10.1128/JCM.38.10.3860-3863.2000google scholar: lookup
          17. Dobbs LJ, Madigan MN, Carter AB, Earls L. Use of FTA gene guard filter paper for the storage and transportation of tumor cells for molecular testing.. Arch Pathol Lab Med 2002 Jan;126(1):56-63.
            pubmed: 11800648doi: 10.5858/2002-126-0056-UOFGGFgoogle scholar: lookup
          18. Mahama CI, Desquesnes M, Dia ML, Losson B, De Deken R, Geerts S. A cross-sectional epidemiological survey of bovine trypanosomosis and its vectors in the Savelugu and West Mamprusi districts of northern Ghana.. Vet Parasitol 2004 Jun 10;122(1):1-13.
            pubmed: 15158552doi: 10.1016/j.vetpar.2004.03.009google scholar: lookup
          19. Allred DR. Antigenic variation in babesiosis: is there more than one 'why'?. Microbes Infect 2001 May;3(6):481-91.
            pubmed: 11377210doi: 10.1016/s1286-4579(01)01404-6google scholar: lookup
          20. Bashiruddin JB, Cammà C, Rebêlo E. Molecular detection of Babesia equi and Babesia caballi in horse blood by PCR amplification of part of the 16S rRNA gene.. Vet Parasitol 1999 Jul;84(1-2):75-83.
            pubmed: 10435792doi: 10.1016/s0304-4017(99)00049-7google scholar: lookup
          21. Hsiao KM, Lin HM, Pan H, Li TC, Chen SS, Jou SB, Chiu YL, Wu MF, Lin CC, Li SY. Application of FTA sample collection and DNA purification system on the determination of CTG trinucleotide repeat size by PCR-based Southern blotting.. J Clin Lab Anal 1999;13(4):188-93.
            pubmed: 10414599doi: 10.1002/(sici)1098-2825(1999)13:4<188::aid-jcla8>3.0.co;2-ggoogle scholar: lookup
          22. Nishimura N, Nakayama T, Tonoike H, Kojima K, Kato S. Direct polymerase chain reaction from whole blood without DNA isolation.. Ann Clin Biochem 2000 Sep;37 ( Pt 5):674-80.
            pubmed: 11026521doi: 10.1258/0004563001899726google scholar: lookup
          23. Holbrook AA. Biology of equine piroplasmosis.. J Am Vet Med Assoc 1969 Jul 15;155(2):453-4.
            pubmed: 5816130

          Citations

          This article has been cited 5 times.
          1. Salinas-Estrella E, Ueti MW, Lobanov VA, Castillo-Payró E, Lizcano-Mata A, Badilla C, Martínez-Ibáñez F, Mosqueda J. Serological and molecular detection of Babesia caballi and Theileria equi in Mexico: A prospective study. PLoS One 2022;17(3):e0264998.
            doi: 10.1371/journal.pone.0264998pubmed: 35259206google scholar: lookup
          2. Tirosh-Levy S, Gottlieb Y, Fry LM, Knowles DP, Steinman A. Twenty Years of Equine Piroplasmosis Research: Global Distribution, Molecular Diagnosis, and Phylogeny. Pathogens 2020 Nov 8;9(11).
            doi: 10.3390/pathogens9110926pubmed: 33171698google scholar: lookup
          3. Guidi E, Pradier S, Lebert I, Leblond A. Piroplasmosis in an endemic area: analysis of the risk factors and their implications in the control of Theileriosis and Babesiosis in horses. Parasitol Res 2015 Jan;114(1):71-83.
            doi: 10.1007/s00436-014-4161-9pubmed: 25280516google scholar: lookup
          4. Odongo DO, Sunter JD, Kiara HK, Skilton RA, Bishop RP. A nested PCR assay exhibits enhanced sensitivity for detection of Theileria parva infections in bovine blood samples from carrier animals. Parasitol Res 2010 Jan;106(2):357-65.
            doi: 10.1007/s00436-009-1670-zpubmed: 19902251google scholar: lookup
          5. Sultan DM, Khalil MM, Abdouh AS, Doleh WF, Al Muthanna AA. Imported malaria in United Arab Emirates: evaluation of a new DNA extraction technique using nested PCR. Korean J Parasitol 2009 Sep;47(3):227-33.
            doi: 10.3347/kjp.2009.47.3.227pubmed: 19724695google scholar: lookup
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