Poliovirus Detection and Molecular Caracterization

The wild poliovirus epidemic and the spread of vaccine-derived poliovirus in October 2019
Global public health efforts are about to eradicate poliomyelitis by immunizing each child until the transmission of the virus is stopped. Poliomyelitis is caused by poliovirus, which is a positive strand non-enveloped virus that occurs in three distinct serotypes (poliovirus 1, poliovirus 2, and poliovirus 3).

pcr

Reference details

Description:
Isolation and molecular characterization of viruses by RT-PCR in real time and RT-PCR and sequencing.
Category of Analysis: Isolation and Genotyping
Pathogen: Poliovirus serotypes 1 to 3
Poliomyelitis with clinical manifestations ranging from non-specific benign disease characterized by fever, sore throat or gastrointestinal symptoms to aseptic meningitis

Specimen

Stool – The type of sample required for poliovirus detectionpoliovirus detection because it is the one in which the poliovirus is most likely to be isolated. To increase the probability of poliovirus isolation, take at least two stool specimens 24 hours apart in patients suspected of having polio. These samples should ideally be collected within 14 days of onset of symptoms.

Zika Virus in vitro diagnostic (IVDs)
Zika Virus Biosafety

Stool – Collect within 14 days of onset of illness and place in a sterile, sealed container, no special media required. Isolation rates increase when two samples are collected 24 hours apart.

Store samples as soon as possible after collection at ≤ -20 ° C until shipment for testing. Ship frozen samples on dry ice. Pack samples of suspected Poliovirus in a shipping container.

Zika virus shipping specimens
Zika Virus Biosafety

Transportation of dangerous goods

Shipment of samples must be made to a person holding a TDG training certificate, as per the TDG Regulations.

Criteria for patients

Cases of acute flaccid paralysis in children under 15 years and suspected cases of poliomyelitis in patients of any age.

Human Antibody
cell-culture

Virus isolation in cell culture

Stool specimens treated for suspected poliomyelitis or acute flaccid paralysis are inoculated into two and RD. Positive isolates of cell lines, L20B and RD. which may be the polio virus, are then typed by intratypic differentiation (DIT) and real-time PCR for VDPV.

Typing of poliovirus isolates and intratypic differentiation

Molecular reagents specific to poliovirus groups and serotypes and vaccine strains and which target characteristic properties of polioviruses in the capsid region (VP1) are used in a series of real-time RT-PCR assays to determine serotype and to distinguish Sabin-like polioviruses from wild polioviruses (intratypic differentiation). When a Sabin virus is identified by DIT, the sample is subjected to a real-time PCR for VDPVs; if the result of the DIT does not correspond to a Sabin-like PV, the sample is routed for RT-PCR and sequencing of the VP1 region to confirm the identity of the virus.

western-blot
Elisa required material

Sabin-type poliovirus analysis for vaccine-derived polioviruses (VDPVs)

A real-time RT-PCR technique targeting areas particularly favorable to the “hot spots” of the capsid region Usually reverting to VDPVs is used to identify possible VDPVs once Sabin-like viruses have been detected by the DIT methods. When a Sabin-type VDPV is identified in a sample, the laboratory report indicates this; when a non-Sabin type VDPV is identified, the sample is routed for RT-PCR and sequencing of the VP1 region to confirm the identity of the virus.

Tommy Ounas

Tommy Ounas

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