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Iranian Journal of Biotechnology، جلد ۶، شماره ۴، صفحات ۲۲۲-۲۲۸
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عنوان انگلیسی 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
چکیده انگلیسی مقاله 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.
کلیدواژه‌های انگلیسی مقاله Coxsackievirus B3,NASBA,RT-PCR
نویسندگان مقاله علیرضا سعیدی نیا |
department of biotechnology, maleke ashtar university, p.o. box 15875-1774, tehran, i.r. iran

مهدی شمس آرا |
department of animal biotechnology, national institute of genetic engineering and biotechnology, p.o. box 14965 161, tehran, i.r. iran
سازمان اصلی تایید شده: پژوهشگاه ملی مهندسی ژنتیک و زیست فناوری

مهدی زین الدینی |
department of biotechnology, maleke ashtar university, p.o. box 15875-1774, tehran, i.r. iran

وحید صادقی |
department of biotechnology, maleke ashtar university, p.o. box 15875-1774, tehran, i.r. iran

نادر مقصودی |
neuroscience research center, shahid beheshti medical university, p.o. box 16765-3718, tehran, i.r. iran

نشانی اینترنتی http://www.ijbiotech.com/article_7041_c62751bfbf62526834d2a0a9c3de9db6.pdf
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