FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / J Vet Med Sci / Development of a loop-mediated isothermal amplification meth...
The Journal of veterinary medical science2014; 76(9); 1271-1275; doi: 10.1292/jvms.14-0140

Development of a loop-mediated isothermal amplification method for detecting Streptococcus equi subsp. zooepidemicus and analysis of its use with three simple methods of extracting DNA from equine respiratory tract specimens.

Abstract: Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is a dominant pathogenic bacterium in equine pneumonia. We developed a specific loop-mediated isothermal amplification (LAMP) method, which targets the gene encoding sorbitol-6-phosphate 2-dehydrogenase (sorD), for detecting S. zooepidemicus and examined the clinical efficacies of its use in combination with each of 3 DNA extraction methods easily used by veterinary practitioners, namely the Loopamp PURE DNA Extraction Kit, InstaGene Matrix and a conventional boiling method. The LAMP method plus the Loopamp PURE DNA Extraction Kit gave higher rates of positivity than the other combinations in both clinical and spiked samples containing clinically significant concentrations (>1 × 10(4) CFU/ml) of S. zooepidemicus.
Get Full Text
Publication Date: 2014-05-28 PubMed ID: 24871644PubMed Central: PMC4197157DOI: 10.1292/jvms.14-0140Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

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 research focuses on the development of a specific method, known as loop-mediated isothermal amplification (LAMP), for detecting a primary cause of pneumonia in horses, Streptococcus equi subsp. zooepidemicus. The study also investigates the effectiveness of this method combined with three different ways of extracting DNA.

Underlying Research Objectives

  • The primary aim of this research is to develop a loop-mediated isothermal amplification (LAMP) method specifically for the detection of Streptococcus equi subsp. zooepidemicus, a primary pathogenic bacterium causing pneumonia in horses.
  • Furthermore, the study intends to evaluate the effectiveness of this new method in conjunction with three easily applied DNA extraction procedures that are usable by veterinary practitioners. These procedures are the Loopamp PURE DNA Extraction Kit, InstaGene Matrix, and a conventional boiling method.

Research Method and Process

  • The LAMP method developed in the study targets the gene encoding sorbitol-6-phosphate 2-dehydrogenase (sorD) – a specific gene identified in S. zooepidemicus.
  • The effectiveness of this method was evaluated both in clinical applications and theoretical samples spiked with significantly high concentrations (>1 × 10(4) CFU/ml) of S. zooepidemicus.
  • Comparisons of the results were made between the LAMP method combined with each of the three DNA extraction techniques.

Findings and Conclusion

  • The outcome of the research revealed that the combination of the LAMP method and the Loopamp PURE DNA Extraction Kit resulted in higher positivity rates for the detection of S. zooepidemicus. This was the case for both actual clinical specimens and spiked samples containing significantly high concentrations of the bacteriophage.
  • This suggests that this particular combination may prove the most effective strategy for detecting the bacterium in a veterinary practice setting.

Cite This Article

APA
Kinoshita Y, Niwa H, Katayama Y. (2014). Development of a loop-mediated isothermal amplification method for detecting Streptococcus equi subsp. zooepidemicus and analysis of its use with three simple methods of extracting DNA from equine respiratory tract specimens. J Vet Med Sci, 76(9), 1271-1275. https://doi.org/10.1292/jvms.14-0140

Publication

The Journal of veterinary medical science
ISSN: 1347-7439
NlmUniqueID: 9105360
Country: Japan
Language: English
Volume: 76
Issue: 9
Pages: 1271-1275

Researcher Affiliations

Kinoshita, Yuta
  • Microbiology Division, Epizootic Research Center, Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotuke, Tochigi 329-0412, Japan.
Niwa, Hidekazu
    Katayama, Yoshinari

      MeSH Terms

      • Animals
      • Colony Count, Microbial / veterinary
      • DNA, Bacterial / chemistry
      • DNA, Bacterial / genetics
      • Horse Diseases / diagnosis
      • Horse Diseases / microbiology
      • Horses
      • Limit of Detection
      • Nucleic Acid Amplification Techniques / methods
      • Nucleic Acid Amplification Techniques / veterinary
      • Pneumonia / diagnosis
      • Pneumonia / microbiology
      • Pneumonia / veterinary
      • Polymerase Chain Reaction / veterinary
      • Sensitivity and Specificity
      • Streptococcal Infections / diagnosis
      • Streptococcal Infections / microbiology
      • Streptococcal Infections / veterinary
      • Streptococcus equi / genetics
      • Streptococcus equi / isolation & purification

      References

      This article includes 26 references
      1. Alber J, El-Sayed A, Lämmler C, Hassan AA, Weiss R, Zschöck M. Multiplex polymerase chain reaction for identification and differentiation of Streptococcus equi subsp. zooepidemicus and Streptococcus equi subsp. equi.. J Vet Med B Infect Dis Vet Public Health 2004 Dec;51(10):455-8.
        doi: 10.1111/j.1439-0450.2004.00799.xpubmed: 15606870google scholar: lookup
      2. Bannister MF, Benson CE, Sweeney CR. Rapid species identification of group C streptococci isolated from horses.. J Clin Microbiol 1985 Apr;21(4):524-6.
        pmc: PMC271711pubmed: 3988897doi: 10.1128/jcm.21.4.524-526.1985google scholar: lookup
      3. Baselski VS, Wunderink RG. Bronchoscopic diagnosis of pneumonia.. Clin Microbiol Rev 1994 Oct;7(4):533-58.
        pmc: PMC358340pubmed: 7834604doi: 10.1128/cmr.7.4.533google scholar: lookup
      4. Båverud V, Johansson SK, Aspan A. Real-time PCR for detection and differentiation of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus.. Vet Microbiol 2007 Oct 6;124(3-4):219-29.
        doi: 10.1016/j.vetmic.2007.04.020pubmed: 17531409google scholar: lookup
      5. Casagrande Proietti P, Bietta A, Coppola G, Felicetti M, Cook RF, Coletti M, Marenzoni ML, Passamonti F. Isolation and characterization of β-haemolytic-Streptococci from endometritis in mares.. Vet Microbiol 2011 Aug 26;152(1-2):126-30.
        doi: 10.1016/j.vetmic.2011.04.009pubmed: 21570219google scholar: lookup
      6. Christley RM, Hodgson DR, Rose RJ, Wood JL, Reids SW, Whitear KG, Hodgson JL. A case-control study of respiratory disease in Thoroughbred racehorses in Sydney, Australia.. Equine Vet J 2001 May;33(3):256-64.
        doi: 10.2746/042516401776249796pubmed: 11352347google scholar: lookup
      7. Fulde M, Valentin-Weigand P. Epidemiology and pathogenicity of zoonotic streptococci.. Curr Top Microbiol Immunol 2013;368:49-81.
        pubmed: 23192319doi: 10.1007/82_2012_277google scholar: lookup
      8. Gotoh K, Nishimura N, Ohshima Y, Arakawa Y, Hosono H, Yamamoto Y, Iwata Y, Nakane K, Funahashi K, Ozaki T. Detection of Mycoplasma pneumoniae by loop-mediated isothermal amplification (LAMP) assay and serology in pediatric community-acquired pneumonia.. J Infect Chemother 2012 Oct;18(5):662-7.
        doi: 10.1007/s10156-012-0388-5pubmed: 22370920google scholar: lookup
      9. Holden MT, Heather Z, Paillot R, Steward KF, Webb K, Ainslie F, Jourdan T, Bason NC, Holroyd NE, Mungall K, Quail MA, Sanders M, Simmonds M, Willey D, Brooks K, Aanensen DM, Spratt BG, Jolley KA, Maiden MC, Kehoe M, Chanter N, Bentley SD, Robinson C, Maskell DJ, Parkhill J, Waller AS. Genomic evidence for the evolution of Streptococcus equi: host restriction, increased virulence, and genetic exchange with human pathogens.. PLoS Pathog 2009 Mar;5(3):e1000346.
        doi: 10.1371/journal.ppat.1000346pmc: PMC2654543pubmed: 19325880google scholar: lookup
      10. Hong CB, Donahue JM, Giles RC Jr, Petrites-Murphy MB, Poonacha KB, Roberts AW, Smith BJ, Tramontin RR, Tuttle PA, Swerczek TW. Etiology and pathology of equine placentitis.. J Vet Diagn Invest 1993 Jan;5(1):56-63.
        doi: 10.1177/104063879300500113pubmed: 8466982google scholar: lookup
      11. Kern M, Böhm S, Deml L, Wolf H, Reischl U, Niller HH. Inhibition of Legionella pneumophila PCR in respiratory samples: a quantitative approach.. J Microbiol Methods 2009 Nov;79(2):189-93.
        pubmed: 19747952doi: 10.1016/j.mimet.2009.08.020google scholar: lookup
      12. Laus F, Attili AR, Cerquetella M, Spaterna A, Tesei B, Cuteri V. Endoscopic findings, microbiological and cytological evaluation of tracheal aspirates in a population of Standardbred horses with poor performances. Vet. Med. (Praha) 2009;54:444–450.
      13. Lavoie JP, Fiset L, Laverty S. Review of 40 cases of lung abscesses in foals and adult horses.. Equine Vet J 1994 Sep;26(5):348-52.
        doi: 10.1111/j.2042-3306.1994.tb04401.xpubmed: 7988535google scholar: lookup
      14. McCue PM, Wilson WD. Equine mastitis--a review of 28 cases.. Equine Vet J 1989 Sep;21(5):351-3.
        doi: 10.1111/j.2042-3306.1989.tb02687.xpubmed: 2776721google scholar: lookup
      15. Merk S, Neubauer H, Meyer H, Greiser-Wilke I. Comparison of different methods for the isolation of Burkholderia cepacia DNA from pure cultures and waste water.. Int J Hyg Environ Health 2001 Nov;204(2-3):127-31.
        doi: 10.1078/1438-4639-00083pubmed: 11759155google scholar: lookup
      16. Merk S, Meyer H, Greiser-Wilke I, Sprague LD, Neubauer H. Detection of Burkholderia cepacia DNA from artificially infected EDTA-blood and lung tissue comparing different DNA isolation methods.. J Vet Med B Infect Dis Vet Public Health 2006 Aug;53(6):281-5.
        doi: 10.1111/j.1439-0450.2006.00956.xpubmed: 16907960google scholar: lookup
      17. Nemoto M, Ohta M, Tsujimura K, Bannai H, Yamanaka T, Kondo T, Matsumura T. Direct detection of equine herpesvirus type 1 DNA in nasal swabs by loop-mediated isothermal amplification (LAMP).. J Vet Med Sci 2011 Sep;73(9):1225-7.
        doi: 10.1292/jvms.11-0065pubmed: 21551979google scholar: lookup
      18. Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T. Loop-mediated isothermal amplification of DNA.. Nucleic Acids Res 2000 Jun 15;28(12):E63.
        doi: 10.1093/nar/28.12.e63pmc: PMC102748pubmed: 10871386google scholar: lookup
      19. Oikawa M, Takagi S, Anzai R, Yoshikawa H, Yoshikawa T. Pathology of equine respiratory disease occurring in association with transport.. J Comp Pathol 1995 Jul;113(1):29-43.
        doi: 10.1016/S0021-9975(05)80066-0pmc: PMC7130345pubmed: 7490335google scholar: lookup
      20. Opel KL, Chung D, McCord BR. A study of PCR inhibition mechanisms using real time PCR.. J Forensic Sci 2010 Jan;55(1):25-33.
        doi: 10.1111/j.1556-4029.2009.01245.xpubmed: 20015162google scholar: lookup
      21. Priestnall S, Erles K. Streptococcus zooepidemicus: an emerging canine pathogen.. Vet J 2011 May;188(2):142-8.
        doi: 10.1016/j.tvjl.2010.04.028pmc: PMC7110628pubmed: 20570190google scholar: lookup
      22. Raidal SL. Equine pleuropneumonia.. Br Vet J 1995 May-Jun;151(3):233-62.
        doi: 10.1016/S0007-1935(95)80175-8pubmed: 7640954google scholar: lookup
      23. Soedarmanto I, Pasaribu FH, Wibawan IW, Lämmler C. Identification and molecular characterization of serological group C streptococci isolated from diseased pigs and monkeys in Indonesia.. J Clin Microbiol 1996 Sep;34(9):2201-4.
        pmc: PMC229217pubmed: 8862585doi: 10.1128/jcm.34.9.2201-2204.1996google scholar: lookup
      24. Sweeney CR, Divers TJ, Benson CE. Anaerobic bacteria in 21 horses with pleuropneumonia.. J Am Vet Med Assoc 1985 Oct 1;187(7):721-4.
        pubmed: 4055490
      25. Timoney JF. The pathogenic equine streptococci.. Vet Res 2004 Jul-Aug;35(4):397-409.
        doi: 10.1051/vetres:2004025pubmed: 15236673google scholar: lookup
      26. Wilson IG. Inhibition and facilitation of nucleic acid amplification.. Appl Environ Microbiol 1997 Oct;63(10):3741-51.
        pmc: PMC168683pubmed: 9327537doi: 10.1128/aem.63.10.3741-3751.1997google scholar: lookup

      Citations

      This article has been cited 6 times.
      1. Garner C, Stephen C, Pant SD, Ghorashi SA. Comparison of PCR-HRM, colorimetric LAMP and culture based diagnostic assays in the detection of endometritis caused by Streptococcus equi subsp. zooepidemicus in mares. Vet Res Commun 2023 Jun;47(2):495-509.
        doi: 10.1007/s11259-022-10047-0pubmed: 36538151google scholar: lookup
      2. Uchida-Fujii E, Kinoshita Y, Niwa H, Maeda T, Nukada T, Ueno T. High prevalence of Mycoplasma equirhinis in Thoroughbred horses with respiratory symptoms in autumn 2018. J Vet Med Sci 2021 Dec 9;83(12):1907-1912.
        doi: 10.1292/jvms.21-0163pubmed: 34732605google scholar: lookup
      3. Jelocnik M, Nyari S, Anstey S, Playford N, Fraser TA, Mitchell K, Blishen A, Pollak NM, Carrick J, Chicken C, Jenkins C. Real-time fluorometric and end-point colorimetric isothermal assays for detection of equine pathogens C. psittaci and equine herpes virus 1: validation, comparison and application at the point of care. BMC Vet Res 2021 Aug 19;17(1):279.
        doi: 10.1186/s12917-021-02986-8pubmed: 34412635google scholar: lookup
      4. Inoshima Y, Takasu M, Ishiguro N. Establishment of an on-site diagnostic procedure for detection of orf virus from oral lesions of Japanese serows (Capricornis crispus) by loop-mediated isothermal amplification. J Vet Med Sci 2017 Jan 10;78(12):1841-1845.
        doi: 10.1292/jvms.16-0268pubmed: 27628591google scholar: lookup
      5. Wickhorst JP, Hassan AA, Sammra O, Huber-Schlenstedt R, Lämmler C, Prenger-Berninghoff E, Timke M, Abdulmawjood A. Phenotypic and genotypic approach to characterize Arcanobacterium pluranimalium isolated from bovine milk samples. Folia Microbiol (Praha) 2016 Sep;61(5):405-12.
        doi: 10.1007/s12223-016-0450-2pubmed: 26883140google scholar: lookup
      6. Knox A, Beddoe T. Enhancement of loop-mediated isothermal amplification (LAMP) with guanidine hydrochloride for the detection of Streptococcus equi subspecies equi (Strangles). PeerJ 2024;12:e17955.
        doi: 10.7717/peerj.17955pubmed: 39421427google scholar: lookup
      Subscribe to our newsletter
      Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.