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:: Volume 9, Issue 1 (2-2015) ::
Iran J Virol 2015, 9(1): 1-6 Back to browse issues page
Development of qRT-PCR Test for Quantification of Rubella Virus in Commercially Available Vaccines
F Sheikholeslami , H Khanahmadi , M Ajorloo , R Ahangari Cohan , Z Karimi , D Norouzian
Pilot Nano biotechnology Department, Pasteur Institute of Iran, Tehran, Iran
Abstract:   (1695 Views)

vaccines is considered as a critical point and in-process of quality control (IPQC) test of vaccine production. Rapid tests, like real time PCR, are more appropriated when the production occurs at industrial scale because of the amounts of starting materials and the excess of consumed time required. In the current study, a real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was developed for Rubella virus (RV) and tested on a commercially available MMR vaccine.

Materials and Methods: The primers and Taq Man probe were designed by Gene runner version 5.0.63 software. Concentrations, as well as reaction temperatures were optimized to establish an efficient qRT-PCR assay for RV RNA. A RV-specific PCR amplicon (109 bp), conserved in all species, was made as an external standard to evaluate the linearity, amplification efficiency, analytical sensitivity and reproducibility of the qRT-PCR.

Results: The real time quantitative assay was shown to have good linearity (R2=0.998), high amplification efficiency (E=96.18%), and high sensitivity (7 × 10 2 copies / 7 µl) for tested vaccine.

Conclusion: The established qRT-PCR method is a simple, rapid, quantitative, highly specific and sensitive assay for quantification of RV RNA copy numbers in IPQC tests at industrial scale.

Keywords: Rubella Virus, MMR Vaccine, Quantitative Real-Time reverse transcription-polymerase chain reaction
Full-Text [PDF 128 kb]   (727 Downloads)    
Type of Study: Original article | Subject: Special
Received: 2016/04/26 | Accepted: 2016/04/26 | Published: 2016/04/26
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Sheikholeslami F, Khanahmadi H, Ajorloo M, Ahangari Cohan R, Karimi Z, Norouzian D. Development of qRT-PCR Test for Quantification of Rubella Virus in Commercially Available Vaccines. Iran J Virol. 2015; 9 (1) :1-6
URL: http://journal.isv.org.ir/article-1-182-en.html


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