Author/Authors :
Nejati, A Polio National Laboratory - Department of Virology - School of Public Health - Tehran University of Medical Sciences , Tabatabaie, H Polio National Laboratory - Department of Virology - School of Public Health - Tehran University of Medical Sciences , Yousefi, M Department of Virology - International Campus - Tehran University of Medical Sciences , Mollaei-Kandelous, Y Immunology Department - Iran University of Medical Science, Tehran , Shahmahmoodi, Sh Food Microbiology Research Center - Tehran University of Medical Sciences
Abstract :
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.
