عنوان به زبان ديگر :
Evaluation of nucleic acid sequence based amplification (NASBA) and Reverse Transcription Polymerase Chain Reaction for detection of coxsackievirus B3 in cell culture and animal tissue samples
پديد آورندگان :
Saeedinla Alireza نويسنده , Shamsara Mehdi نويسنده , Zeinoddini Mehdi نويسنده , Sadeghi Vahid نويسنده , Maghsoudi Nader نويسنده
چكيده لاتين :
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 KCI concentration. The optimal concentration
of KCI was determined as 90 mM. Serial dilutions of 1 mo g 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.
