Assay for identification of heterozygous single-nucleotide polymorphism (Ala67Thr) in human poliovirus receptor gene.
OBJECTIVE
It is important to understand the role of cell surface receptors in susceptibility to infectious diseases. CD155 a member of the immunoglobulin super family, serves as the poliovirus receptor (PVR). Heterozygous (Ala67Thr) polymorphism in CD155 has been suggested as a risk factor for paralytic outcome of poliovirus infection. The present study pertains to the development of a screening test to detect the single nucleotide (SNP) polymorphism in the CD155 gene.


METHODS
New primers were designed for PCR, sequencing and SNP analysis of Exon2 of CD155 gene. DNAs extracted from either whole blood (n=75) or cells from oral cavity (n=75) were used for standardization and validation of the SNP assay. DNA sequencing was used as the gold standard method.


RESULTS
A new SNP assay for detection of heterozygous Ala67Thr genotype was developed and validated by testing 150 DNA samples. Heterozygous CD155 was detected in 27.33 per cent (41/150) of DNA samples tested by both SNP detection assay and sequencing.


CONCLUSIONS
The SNP detection assay was successfully developed for identification of Ala67Thr polymorphism in human PVR/CD155 gene. The SNP assay will be useful for large scale screening of DNA samples.
Enterovirus Research Centre (ICMR), Mumbai, India.
Shyam Sundar
SS
Nandi
Enterovirus Research Centre (ICMR), Mumbai, India.
Deepa Kailash
DK
Sharma
Enterovirus Research Centre (ICMR), Mumbai, India.
Jagadish M
JM
Deshpande
D005838:Genotype; D006579:Heterozygote; D006801:Humans; D011051:Poliomyelitis / Q000175:diagnosis / Q000235:genetics*; D017955:Poliovirus / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide; D011991:Receptors, Virus / Q000235:genetics* / Q000302:isolation & purification; D017422:Sequence Analysis, DNA / Q000379:methods*
D011991:Receptors, Virus; C066100:poliovirus receptor