Author/Authors :
-، - نويسنده Department of Biotechnology, Maleke Ashtar University, P.O. Box 15875-1774, Tehran, I.R. Iran Saeedinia, Alireza , -، - نويسنده Department of Animal Biotechnology, National Institute of Genetic Engineering and Biotechnology, P.O. Box 14965/161, Tehran, I.R. Iran Shamsara, Mehdi , -، - نويسنده Department of Biotechnology, Maleke Ashtar University, P.O. Box 15875-1774, Tehran, I.R. Iran Zeinoddini, Mehdi , -، - نويسنده Department of Biotechnology, Maleke Ashtar University, P.O. Box 15875-1774, Tehran, I.R. Iran Sadeghi, Vahid , -، - نويسنده Neuroscience Research Center, Shahid Beheshti Medical University, P.O. Box 16765-3718, Tehran, I.R. Iran Maghsoudi, Nader
Abstract :
Enteroviruses are the causative agents of a number of diseases in humans. Group B coxsackieviruses are believed to be the most common viral agents responsible for human heart disease. Genomic data of enteroviruses has allowed developing new molecular approaches such as Nucleic Acid Sequence Based Amplification (NASBA) for detection of such viruses. In this study, coxsackievirus B3 (CVB3) was detected in virus-infected cell culture and specimens of artificially infected mice with specific primers using Reverse Transcription - Polymerase Chain Reaction (RT-PCR) and NASBA techniques. According to the results, both techniques could be used for the detection of viruses in cell culture and artificially infected animals. NASBA reaction was simpler to perform than RT-PCR. The only variable factor that had to be optimized with NASBA is KCl concentration. The optimal concentration of KCl was determined as 90 mM. Serial dilutions of 1 mg of total RNA showed that both RT-PCR and NASBA could detect the virus at 10-5 dilution. Analyses of heart and spleen samples from infected animals were positive for presence of Coxsackievirus B3 with both RT-PCR and NASBA. In conclusion, NASBA offers some advantages over RT-PCR and is a suitable alternative technique for the sensitive detection of CVB3 in contaminated samples.
