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.

CVB3, Myocaditis, RNA extraction, RT-PCR