rt-pcr detection of coxsackievirus b3: a viral myocarditis

backgrounds and aims: coxsakievirus b3 (cvb3), one of the six coxsakievirus b serotypes, is a member of the enterovirus genus within the picornaviridae family. cvb3 is an important pathogen of viral myocarditis, which accounts for more than 50% of viral myocarditis cases. the genome of cvb3, like that of other entroviruses, is a single-stranded, sense, polyadenylated rna molecule with 7400 nucleotides in length and a single open reading frame (orf), flanked by 5΄ and 3΄ non-translated regions. the capsids of coxcakieviruses are composed of the four structural proteins: viral protein-1 (vp1), vp2, vp3, and vp4. in the present study, a new set of primers were designed based on the vp1 for rt-pcr detection of cvb3. materials and methods: total rna was extracted from cvb3-infected hela cell line and cdna was synthesized using random primers. then, pcr was carried out by specific primers and the pcr product analysis was performed using 1% agarose gel electrophoresis. moreover, the sensitivity and specificity of this method were determined using serial dilution of cvb3 cdna and three genuses of entroviruses, respectively. results: rt-pcr assay revealed a 234 bp specific amplified fragment. the sensitivity of this test was determined 5.72 fg/µl cdna. on the other hand, the specificity was successful in comparison with coxsackievirus a16, echovirus 36 and rhinovirus. conclusions: the rt-pcr is a highly sensitive and rapid technique for detecting cvb3 infection. moreover, this method can be used as an easy diagnostic test in regard with cvb3 detection in the clinical laboratories.