Human Enteroviruses (HEVs), the highly dominant enteric pathogens that affect millions of people worldwide each year, which are small non enveloped spherical- shaped viruses and one of Picornaviridae family genera include polioviruses (PVs) and non-polio enteroviruses (NPEVs) (coxsackieviruses, echoviruses, and other new NPEVs) [1]. Their genome is made up of a positive-sense, single-stranded RNA includes a 5′ and 3′ noncoding region squeezed in a single open reading frame. The polyprotein is co- and post-translational cleaved to yield four structural proteins: VP4, VP2, VP3, and VP1 [2].

Molecularly, according to the partial or complete HEVs nucleotide sequences analysis (VPg+5′UTR), HEVs are classified into four species: HEV-A (24 types), HEV-B (61 types), EV-C (23 types involving Poliovirus), and 5 types to HEV-D [3]. In the last few decades, the Non-Polio Enteroviruses showed high concern with a wide range of diseases worldwide such as hand, foot, and mouth disease, aseptic meningitis, encephalitis, type I diabetes mellitus, myocarditis, neuromuscular diseases and meningitidis [4,5] and revealed a serious problem of Acute Flaccid Paralysis (AFP) in children aged younger than 15 years [6].

Whith high hands of the routine work of the poliovirus Iraqi national laboratory in the detection of AFP associated- enteroviruses (PVs and NPEVs) by the tissue culture method. The increasingly serious problems of the acute flaccid paralysis that reflected in the residual paralysis cases and even deaths among Iraqi children are increasingly reported. There is only one study before, isolated the NPEVs from patients with AFP by microneutralization method [7]. And no even one single molecular study in Iraq for the AFP associated- NPEVs. As a result, there was a critical need to establish a study aim to (i) detect the tissue culture NPEVs' positive and negative isolates from the stool samples of children with AFP by the RT- PCR using 5′- non coding region primer to overcome the tissue culture problems, As no single cell line is sufficient to isolate of all serotypes, lack of sensitivity for some cell lines, slow often requiring 3 to 6 days for CPE to appear, limited by low viral titers in some samples, and the propability of getting false negative and weak positive results beside the consumed effeorts and time [8]. (ii) to displace the microneutralization technique that may give interrupted results due to the high mutational rate of some viral genomes [9].

(ii) typing the genotypes of the death and residual paralysis, tissue culture false negative and weak positive associated-NPEVs isolates by the nucleotide sequence analysis to the complete VP4 capsid gene to unfold the relatedness between the genome sequence image (i.e. the polymorphisms effects) of the NPEVs and their effects on the phenotype, structure and their behavior in the tissue culture, clinical features and their fatality rate on these viruses.

And (iii) to reveal out the evolutionary relationship with the convergence ratio (similarity) among the sequenced/ local NPEVs strains in comparison with the reference/ protypes strains that available on the NCBI by the phylogenetic tree and the similarity matrix analysis and document them in the NCBI website.