Recognition of coxsackievirus A by Enterovirus genus-specific immune and molecular markers in experimentally infected suckling mice.
OBJECTIVE
Most of coxsackieviruses A (CV-A) are difficult to isolate in cell culture and are responsible for flask paralysis in suckling mice. The aim of the present work was to analyze the ability of immune and RT-PCR techniques to detect viral components of three different serotypes, CV-A6, CV-A13, and CV-A14, in skeletal muscles of experimentally infected suckling mice.


METHODS
The antigen detection was done by immunofluorescence technique on trypsinized muscular cells and by immunoperoxidase assay on frozen sections of skeletal muscle, using a monoclonal antibody directed towards a conserved epitope of the VP1 capsid protein among enteroviruses. The nested RT-PCR technique used primers located in the 5' non coding region of viral RNA.


RESULTS
The group antigen was present in muscle cells of suckling mice infected by the three serotypes of CV-A which were assayed. Similarly, the muscle specimens were positive by nested RT-PCR. A kinetic study performed with CV-A13 and CV-A14 showed that the RT-PCR assay was positive as soon as 24 h after infection whereas the detection of VP1 antigen and symptoms of flask paralysis were observed only 48 and 72 h after infection, respectively.


CONCLUSIONS
These results show that the tested serotypes of CV-A can be easily detected in muscle specimens of suckling mice by using antigenic and molecular techniques currently available for the diagnosis of enterovirus infections.
Nsaibia
Siwar
S
Laboratory of Transmissible Diseases and of Biologically Active Substances, MDT01, Faculty of Pharmacy, University of Center, Monastir, Tunisia.
Ben Othman
Samia
S
Trabelsi
Abdelhalim
A
Bourlet
Thomas
T
Aouni
Mahjoub
M
Pozzetto
Bruno
B
D000818:Animals; D000833:Animals, Suckling; D003384:Coxsackievirus Infections / Q000821:virology*; D004770:Enterovirus / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D005455:Fluorescent Antibody Technique; D007124:Immunoenzyme Techniques; D051379:Mice; D018482:Muscle, Skeletal / Q000821:virology; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012703:Serotyping
D012367:RNA, Viral