Author/Authors :
Mousavi-Nasab, Dawood Department of Research and Development - Production and Research Complex - Pasteur Institute of, Iran , Sabahi, Farzaneh Department of Virology - Faculty of Medical Sciences - Tarbiat Modares University, Tehran, Iran , Kaghazian, Hooman Department of Research and Development - Production and Research Complex - Pasteur Institute of, Iran , Paryan, Mahdi Department of Research and Development - Production and Research Complex - Pasteur Institute of, Iran , Mirab Samiee, Siamak Reference Health Laboratories Research Microbiology - Karaj Branch Islamic Azad University, Karaj, Iran , Ghaderi, Mostafa Department of Microbiology - Karaj Branch Islamic Azad University, Karaj, Iran , Zali, Fatemeh 5 Department of Clinical Biochemistry - Faculty of Medicine - Tehran University of Medical Sciences, Tehran, Iran , Makvandi, Manoochehr Infectious and Tropical Diseases Research Center - Health research Institute - Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
Abstract :
HRV is the causative agent of severe gastroenteritis in children and responsible for two million hospitalizations and more than a half-million deaths annually. Sequence characteristics of the gene segments encoding the VP7 and VP4 proteins are used for the genotype classification of rotavirus. A wide variety of molecular methods are available, mainly based on reverse transcription PCR for rapid, specific and sensitive genotyping of rotaviruses. This study describes an alternative real-time PCR assay for genotyping of rotavirus. Methods: The samples of stools studied in this research have been collected from patients referred to Children's Medical Centers, Tehran, Iran. Rotavirus detection and genotyping were performed using the RT-PCR and semi-nested RT-PCR, respectively. Samples were then genotyped with a new real-time PCR. Results: The real-time PCR was able to genotype all positive samples with a mean Ct of 28.2. Besides, a concordance rate of 100% was detected between real-time PCR and semi-nested RT-PCR. Conclusion: In this study, the genotyping of rotavirus with real-time PCR showed that this method can provide several favorable features, including high sensitivity and specificity, and a wide dynamic range for rotavirus genotyping.
Keywords :
Gastroenteritis , Genotype , Real-time polymerase chain reaction , Rotavirus
