AffiPCR® Poliovirus Real Time PCR

Target Disease Type: Neurological Infections

Specific Application: Enterovirus/Poliovirus

Storage and Shipping : on request

Poliovirus RT-PCR V58-50FRT DataSheet

INTRODUCTION

Poliomyelitis is an acute paralytic disease caused by poliovirus, a Human Enterovirus C of the Picornaviridae family which is classified into three distinct and stable serotypes. Since the 1960s, poliomyelitis has been effectively controlled by the use of inactivated or live attenuated vaccines. The Sabin live oral poliovirus vaccine (OPV) is constituted of attenuated strains of each of the three serotypes which have been selected by numerous passages of wild-type strains in monkey tissues in vivo and in vitro. The OPV strains (Sabin 1, 2 and 3) replicate in the human gut and induce a strong and long-lasting immune response, including a local intestine immunity. However, in rare cases (1case per 0.2 to 2.5million doses), OPV strains can cause vaccine-associated paralytic poliomyelitis (VAPP) . Reversion of vaccine strains toward a pathogenic phenotype is probably one of the main causes of VAPP, a disease most frequently associated with type Sabin 3 and type Sabin 2 strains and more rarely with the type Sabin 1 strain.

INTENDED USE

Kit Poliovirus Real-TM is a Real-Time test for the qualitative detection of Poliovirus/Enterovirusgroup C (HEV-C) RNA and differentiation of Poliovirus strains Sabin 1, Sabin 2, Sabin 3 in the biological materials and in the environment.

PRINCIPLE OF ASSAY

Kit Poliovirus Real-TM is based on three major processes: isolation of RNA from specimens, reverse transcription of the RNA and Real Time amplification. Test contains the Internal Control (IC) which must be used in the RNA purification procedure and serves as an amplification control for each individually processed specimen and to identify possible reaction inhibition.

Poliovirus detection by the polymerase chain reaction (PCR) is based on the multiplex amplification of the pathogen genome specific region in two tubes using specific primers:

• PCR-mix-1 HEV-C/IC tube contains primers directed against Poliovirus and Enterovirus group C (HEV-C) RNA and Internal Control (HEV-C cDNA is detected in the FAM/Green fluorescence channel and IC is detected in the JOE/Yellow/HEX/Cy3 channel) ;
• PCR-mix-1-FL Sabin 1/2/3 tube contains primers directed against Poliovirus strains Sabin 1, Sabin 2, Sabin 3 (Sabin 1 cDNA is detected in the ROX/Orange/TexasRed fluorescence channel, Sabin 2 cDNA is detected in the FAM/Green channel, Sabin 3cDNA is detected in the JOE/Yellow/HEX/Cy3 channel) .