Identification of RD and L20B Cell Lines Cross-Contamination by Multiplex SYBR-Green PCR in Poliovirus Laboratory

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.
In this study, a multiplex SYBR-Green PCR based on Cytochrome b oxidase amplification was designed to L20B and RD cell lines cross-contamination.
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.
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.