comparative analysis of routine techniques: Reverse transcriptase polymerase chain reaction [RT-PCR] and five cell lines for detection of enteroviruses in stool specimens

ABSTRACT

Each year, Enteroviruses infect millions of people and cause different diseases. The agents are usually detected using cell culture. RD [Rhabdomyosarcoma] and L20B [L cells] are among the recommended cells by the World Health Organisation [WHO] for this purpose. Even though cell culture is the most common method used in diagnosing Enteroviruses in stool specimens, this particular method poses some problems, which include false positive or negative results, lack of a unique cell line for diagnosing all Entero virus types in addition to being time consuming. For these reasons, an attempt was made to find better techniques of Entero virus detection. RT-PCR [Reverse Transcriptase Polymerase Chain Reaction] is a technique used in place of the cell culture method. In this study, the cell culture method was compared with RT-PCR for detection of Enteroviruses in stool specimens. First, the chloroform treated stool samples were inoculated onto five cell lines, including RD, L20B, Hep-2 [Human Epidermoid carcinoma cell line], Vero [Verde Reno] and GMK [Green Monkey Kidney]. The results were then compared with data from Entero virus detection using the RT-PCR technique. The difference between RT-PCR and cell culture results was significant. Enteroviruses were detected in 24% of specimens using RT-PCR while cell lines could isolate Enteroviruses in just 14.4% of the samples