development of qrt-pcr test for quantification of rubella virus in commercially available vaccines
نویسندگان
چکیده
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.
منابع مشابه
Development of qRT-PCR Test for Quantification of Rubella Virus in Commercially Available Vaccines
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...
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عنوان ژورنال:
iranian journal of virologyجلد ۹، شماره ۱، صفحات ۱-۶
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