en عمومى General پژوهشي Original article <div><strong>Background and Aims:</strong> 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.<br> <strong>Materials and Methods:</strong> In this study, a multiplex SYBR-Green PCR based on Cytochrome b oxidase amplification was designed to L20B and RD cell lines cross-contamination. <strong>Results:</strong> 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.<br> <strong>Conclusions: </strong>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.</div> Multiplex SYBR-Green PCR Poliovirus, Cell line authentication 14 18 http://journal.isv.org.ir/browse.php?a_code=A-10-135-1&slc_lang=en&sid=1 A Nejati 10031947532846006677 10031947532846006677 No Polio National Laboratory, Department of Virology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran, telephone number: +982188950595 H Tabatabaeie 10031947532846006678 10031947532846006678 No Polio National Laboratory, Department of Virology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran, telephone number: +982188950595 M Yousefi 10031947532846006679 10031947532846006679 No Polio National Laboratory, Department of Virology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran, telephone number: +982188950595; Department of Virology, International Campus, Tehran University of Medical Sciences, Tehran, Iran. Y Mollaei-Kandelous 10031947532846006680 10031947532846006680 No Polio National Laboratory, Department of Virology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran, telephone number: +982188950595; Immunology Department, Iran University of Medical Science, Tehran 14716-13151, Iran. Sh Shahmahmoodi 10031947532846006681 10031947532846006681 No Polio National Laboratory, Department of Virology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran, telephone number: +982188950595; Food Microbiology Research Center, Tehran University of Medical Sciences, Tehran, Iran.