Expression And Preliminary Diagnostic Application Of HCMV Recombinant Chimeric Antigen

Objective: To obtain HCMV seroprevalence in pregnant women, children and so on inHefei by ELISA three antigen fusion protein, we induce expression, purify target protein andpreliminarily，the strongly and highly hydrophilic DNA fragments, were constructedcomparing the sensitivity and specificity of three recombinant chimeric antigens andanalyze risk factors of HCMV infection in pregnant women and children.Methods: DNA sequences of HCMV including UL32, UL44, UL55, UL83, UL123analyzed by Protean software was connected by Overlap method in introducingBgl II,BamH I restriction sites,which construct three recombinant plasmids and induce toexpress. the recombinant protein which puricated by Ni2+affinity chromatographywas named HCMV pQE80-L-UL32-UL44-UL83, pQE80-L-UL32-UL44-UL-55-UL83,pQE80-L-UL32-UL44-UL-55-UL83-UL123. Immunoreactivity wasdetected byWesternblot.76pregnant women,138children And41patients with serum tumor associatedwith KSHV was tested by the indirect ELISA with thesethree recombinant proteins asantigens. Using SPSS160software to analyze andprocess the resultsResult：The recombinant fusion antigen expression plasmid is constructed to obtain target protei-n. Multi-epitope antigen is confirmed to be the specific antigen for HCHV andhas immunity by Western blot. the detection methods was established by recombinant antig-en the commercial kits sensitivity was97.6%and specificity of100%,but the recombinantantigen both sensitivity and specificity were100%.①The positive rate of pregnant serum HCMV was55.3%, and the infection rateof19to21weeks during gestation was75%. Logistic regression analysis showed thatthere was no significant correlation between the positive rate of HCMV and maternal gestational age, AFP (alpha-fetoprotein), HCG (human chorionic gonadotropin),unconjugated estriol levels (P>0.05).②The positive rate of children serum HCMVwith respiratory diseases was32.7%.Logistic regression analysis showed that there weresignificant correlations (P<0.05) between the positive rate of HCMV and the children’sages, levels of lymphocytes; whereas the positive rate of HCMV that compared withgender, WBC (white blood cells), RBC (red blood cells), PLT (platelets) was nosignificant (P>0.05). And the positive rate of KSHV in these serum was16.7%. Logisticregression analysis showed that there was no significant correlation between children’sage, sex, WBC, RBC, PLT, levels of CRP and the lymphocytes and the positive rate ofKSHV(P>0.05).③The positive rate of HCMV serum with tumors was20.9%, andthere are seven cases in which tumors detected HCMV infection.Logistic regressionanalysis showed that age, gender, tumor and HCMV infection rate were no significantcorrelation(P>0.05).④The positive rate of KSHV serum with tumors was30.2%,Logistic regression analysis showed that tumor with KSHV infection rate weresignificant correlation (P<0.05)The infection rates HCMV and KSHV in150children’s serum were32.7%and16.7%. Both infection rates were statistically significant (χ2=9.93, P=0.002). Thepositive rate of HCMV and KSHV were55.3%and1.3%in76pregnant women, bothpositive rates were statistically significant (χ2=39.09, P=0.000). The positive rate ofHCMV and KSHV in43cancer patients were20.9%and30.2%respectively, and thepositive rates were no significant difference (χ2=1.125, P=0.289).Conclusion：The recombinant fusion antigen expression plasmid is constructed to obtain targetprotein. Multi-epitope antigen is confirmed to be the specific antigen for HCHV and has immunity by Western blot. the detection methods was established by recombinantantigen. the infection rates of HCMV had no differences with gender, age, AFP,HCG and estrogen levels in pregnant women; There were significant correlations between the infection rate of HCMV and ages of10.01to12months and percentage of lymphocytes level that was over40.01in children with MPP. There isclose relationship between HCMV infection and respiratory infection, that because-e of active HCMV infection in children leads to the decline of lung functionandlow immunity occurrence of mycoplasma pneumonia in recurrent respiratory infec-tion; tthe low immunity in children is the result of the virus infection.MP Seco-ndly, the serum of HCMV IgG infection in children with mycoplasma pneumonia-e pneumonia than that of KSHV infection.