Development of a Diagnostic Shuttle PCR Test Targeting the Streptococcus equi SeM Gene

Changes in Serum Antibody Levels after Vaccination for Strangles and after Intranasal Challenge with Streptococcus equi subsp. equi in Horses

“…S. equi was isolated by inoculating Colombian agar containing 5% horse blood with 50 μ l of the suspension, and S. equi was identified using API Strep 20 (SYSMEX bioMerieux, Tokyo, Japan). Total DNA was extracted from the suspension using InstaGene Matrix (BIO-RAD, U.S.A.), and the S. equi seM gene was detected with a semi-nested PCR method [ 1 ]. Additionally, the seM genotype of the isolated S. equi was analyzed as described in a previous report [ 2 ].…”

Development and Application of Loop-Mediated Isothermal Amplification Methods Targeting the <i>seM</i> Gene for Detection of <i>Streptococcus equi</i> subsp. <i>equi</i>

“…The semi-nested PCR targeting the seM gene (seM semi-nested PCR) was performed according to a previous report [1] with some modifications. In brief, initial PCR was performed in a 25 μl volume containing 2.0 μl of template DNA solution, 0.4 mM each of SEf and SEr1 PCR primer (Table 2), 20 mM Tris-HCl (pH 8.0), 100 mM KCl, 3.0 mM MgCl 2 , 0.2 mM each of deoxynucleotide triphosphates and 0.6 U of DNA polymerase (z-Taq, Takara Bio Inc., Shiga, Japan).…”

“…However, the diagnostic procedure conventionally used as a culture examination for S. equi requires significant time before the result is obtained. The alternative PCR method that detects the S. equi gene to confirm the diagnosis genetically can be applied clinically [1,4,11]; however, significant time is required to conduct the PCR method, although the disease can be diagnosed more promptly with PCR than by culture. The development of a rapid diagnostic method is highly desirable to alleviate these delays.…”