Detection Of Human Cytomegalovirus In Gliomas By Drop In/out PCR

Objective:We aimed to investigate accuracy and sensitivity of drop in/out PCR and to validate whether can it be used to detect the HCMV in tumor tissues.Moreover,we investigate the relationship between the infection of HCMV and type of tumors,WHO grades of glioma and cellular receptor PDGFRαand EGFR through the detection of HCMV in 50 human intracranial tumor tissues.Methods:Applying the gene cloning we established cloned plasmid of HCMV,and then undergo gradient dilution.Through testing the plasmid of HCMV of different concentration using drop in/out PCR,we confirmed the lowest concentration of the plasmid that is detectable and then analyze the sensitivity of this method used for detecting HCMV.Then,HCMV in 50 intracranial tumors was tested using this kind of method.The valid results were then detected of HCMV IE in immunohistochemistry.All the two assays’results,as well as the corresponding pathological data,were used for exploring the relationship of the HCMV infection and the type of tumor and WHO grades of astrocytoma.At last,immunohistochemistry was applied again to detect PDGFRαand EGFR of all the tumor tisses which is of valid results in drop in/out PCR,and then indentified the relationship between intensity of HCMV infection and expression volume of PDGFRαand EGFR.Results:In current study,we find that the lowest concentration of HCMV plasmid is5*10~4copies/ml,clarified that it is of proper accuracy for drop in/out PCR to detect the HCMV gene.In the detection of HCMV in tumor tissues by this kind of method,41 of 50valid result was obtained,which contained 29 of 41 positive results.Statistic analysis demonstrated the positive rate is of no obvious differentiation between the results that is obtained by the using of drop in/out PCR(p>0.05)and immunohistochemistry,and the consistency of the results of two different methods is decent(p<0.05).Statistic results also indicates that neither the presence of HCMV in tissues nor intensity of HCMV infection has obvious correlation to the type of HCMV and WHO grades of astrocytoma(p>0.05).Finally,we also proved that there is no distinct correlation between the existence of HCMV infecion and the presence of PDGFRαand EGFR(p>0.05).And we found that the positive correlation between the intensity of HCMV infection and expression quantity of EGFR is obvious(p<0.05),but is not between intensity of HCMV infection and expression quantity of PDGFRα.Conclusions:1.Drop in/out PCR is of good accuracy and sensitivity and can be used to detect the HCMV in tumor tissues.2.Neither the presence of HCMV in tissues nor the intensity of HCMV infection has correlation between the type of tumors and WHO grades of astrocytoma..3.The existence of infections of HCMV in tissues is unobvious correlated to the presence of PDGFRαand EGFR.The intensity of HCMV infection is significantly positively correlated to the expression quantity of EGFR,but not to the expression quantity of PDGFRα.