:: Volume 11, Issue 2 (6-2017) ::
Iran J Virol 2017, 11(2): 14-18 Back to browse issues page
Identification of RD and L20B Cell Lines Cross-Contamination by Multiplex SYBR-Green PCR in Poliovirus Laboratory
Abstract:   (740 Views)
Background and Aims: World Health Organization Global Polio Laboratory Network (GPLN) plays a critical role in the Global Polio Eradication Initiative. Cell culture methods (mostly RD and L20B cell lines) have been used for Enteroviruses and polioviruses isolation, respectively. Cross-contamination among L20B and RD cell lines causes the problem in accuracy of poliovirus surveillance and decreases the poliovirus detection. Therefore, validation of identity of cell lines purity is a vital part of cell culture in polio laboratory.
Materials and Methods: In this study, a multiplex SYBR-Green PCR based on Cytochrome b oxidase amplification was designed to L20B and RD cell lines cross-contamination. Results: The conventional multiplex PCR performed on DNA extracted from L20B cells deliberately cross-contaminated with RD cells clearly showed not only the identity of L20B cell line but also the presence of contaminant RD cells.
Conclusions: The results indicated that the multiplex SYBR-Green PCR was reliable method to identity L20B and RD cell lines individually and also after deliberate cross-contamination.
Keywords: Multiplex SYBR-Green PCR Poliovirus, Cell line authentication
Full-Text [PDF 504 kb]   (454 Downloads)    
Type of Study: Original article | Subject: General
Received: 2017/12/3 | Accepted: 2018/04/15 | Published: 2018/04/15

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Volume 11, Issue 2 (6-2017) Back to browse issues page