Arch Iran Med. 2016;19(9): 0-0.
PMID: 27631177
Scopus ID: 84987653257
  Abstract View: 823
  PDF Download: 779

Original Article

Development of Poly (A)-Tailed Universal Reverse Transcription PCR Method for Sequence-Independent Amplification of Rearranged Rotavirus

Elham Ahmadi, Hoorieh Soleimanjahi * , Majid Sadeghizadeh, Ali Teimoori


BACKGROUND: Group A Rotaviruses (GARV) are the main viral cause of acute gastroenteritis, leading to 870,000 deaths annually in the developing world and representing a major health problem. Therefore, diagnosis and treatment of this disease are crucial. Gene rearrangement within segmented viruses as well as rotavirus is seen throughout chronic rotavirus infection in immunodeficient young children and through serial passage of rotavirus in cell culture at a high multiplicity of infection. Detailed knowledge of rotavirus biology allows design of a vaccine against rotavirus by engineered antigens. The aim of this study was to develop Poly (A) -Tailed universal Reverse Transcription Polymerase Chain Reaction (RT-PCR) method and compare the efficacy of this procedure with specific multiplex PCR protocol for detecting normal and rearranged segments.

METHODS: Virus was propagated on confluent monolayer of MA-104 cells and aliquots of each passage were kept frozen for further RNA genomic profiles analysis by polyacrylamide gel electrophoresis. Purified Rota virus RNA was polyadenylated and used for the amplification and detection of normal and rearranged segments of rotavirus using RT-PCR. 
RESULTS: The generation of gene rearrangement through multiple serial passages of rotavirus was shown using MOI ≥ 1 . The rearranged RNA segments of NSP1 and NSP3 genes with different migration pattern in PAGE were detected by poly(A)RT-PCR.
CONCLUSION: In the current research, a novel Poly(A) -Tailed Universal Reverse Transcription PCR method was introduced for the high throughput amplification and analysis of the informative untranslated regions of the rotavirus genome.
First Name
Last Name
Email Address
Security code

Abstract View: 823

Your browser does not support the canvas element.

PDF Download: 779

Your browser does not support the canvas element.

ePublished: 01 Sep 2016
EndNote EndNote

(Enw Format - Win & Mac)

BibTeX BibTeX

(Bib Format - Win & Mac)

Bookends Bookends

(Ris Format - Mac only)

EasyBib EasyBib

(Ris Format - Win & Mac)

Medlars Medlars

(Txt Format - Win & Mac)

Mendeley Web Mendeley Web
Mendeley Mendeley

(Ris Format - Win & Mac)

Papers Papers

(Ris Format - Win & Mac)

ProCite ProCite

(Ris Format - Win & Mac)

Reference Manager Reference Manager

(Ris Format - Win only)

Refworks Refworks

(Refworks Format - Win & Mac)

Zotero Zotero

(Ris Format - Firefox Plugin)