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

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.