| Objective:In order to understand Kaposi’s Sarcoma-associated Herpesvirus（KSHV）infection in the general population of Hefei and Men who have sex with Men（MSM）in Shanghai,and to understand the risk factors associated with KSHV infection,the positive rate of KSHV was compared between the two regions to provide a strong basis for the control and prevention of KSHV.Methods:（1）The target genes ORF65C,ORF73C and K8.1γwere constructed into the p QE-80L expression vector with His tag,and the recombinant proteins induced in large quantities were purified by the way of competitive binding with Ni-NTA.The purified protein was used as an enzyme-linked immunosorbent antigen to detect KSHV antibodies in serum samples.Finally,the results of the test were combined with statistical patient information and the positive rate of KSHV infection in the two groups for regression analysis using SPSS 26.0,to obtain factors associated with the risk of infection with KSHV.（2）New Zealand white rabbits were immunized six times with three purified recombinant proteins with specific antigenic amounts,twice subcutaneously,and four times by ear vein to obtain polyclonal antibodies.（3）The iSLK.219 cell line containing the KSHV genome was used to amplify the VR1region of its K1 gene by nested PCR,and then establish a genotyping method based on KSHV K1.Results:（1）Three recombinant genes p QE-80L-ORF65,p QE-80L-ORF73 and p QE-80L-K8.1 were successfully constructed,and three recombinant proteins with good purity were purified by Ni-NTA agarose purification resin.（2）Three recombinant proteins were used as antigens to detect KSHV antibodies in serum of patients.A total of 1009 serum samples were collected from Hefei,Anhui,and553 serum samples were collected from Men who have sex with Men（MSM）population in Minhang District,Shanghai.The seroprevalence of KSHV was 11.30%in the population of Hefei,Anhui.In Minhang District of Shanghai,the seroprevalence of KSHV in MSM population was 16.46%.（3）There is a significant difference between the KSHV seroprevalence of the general population and the KSHV seroprevalence of the MSM（95%CI,X~2=8.333,P=0.004）.The regression analysis of the samples showed that diabetes was significantly correlated with KSHV infection（OR=1.578,95%CI 1.072-2.323,P=0.021）,hyperlipidemia was significantly correlated with KSHV infection（OR=2.468,95%CI 1.345-4.526,P=0.004）,the age group of 50 years showed a significant correlation with KSHV infection（OR=2.533,95%CI 1.528-4.199,P<0.001）.However,coronary heart disease,hyperuricemia,hypoproteinemia,gender,hypertension,malignant tumor,HIV,and HBV,occupation,education level,marital status,current status if unmarried,and monthly income were not significantly correlated（P>0.05）.（4）Polyclonal antibodies against ORF65,ORF73 and K8.1 were successfully prepared.The titers of the three polyclonal antibodies were 1:400,1:3200 and 1:1600,respectively,by indirect ELISA method,and three purified proteins were verified to have strong immunogenicity.（5）The KSHV classification method based on K1 VR1 region was established.Conclusion:Through analysis and comparison,it was found that the risk of KSHV infection in the MSM population is greater than that in the general population,and the risk of KSHV infection in people with diabetes（positive rate was 19.15%）is higher than that in people without diabetes（OR=1.578,95%CI 1.072-2.323,P=0.021）.The risk of KSHV infection in patients with hyperlipidemia（positive rate was 28.07%）was higher than that in patients without hyperlipidemia（OR=2.468,95%CI 1.345-4.526,P=0.004）,infection with KSHV in 50 years old group（positive rate was 17.22%）increased the risk of viral infection（OR=2.533,95%CI 1.528-4.199,P<0.001）.It is suggested that diabetes,hyperlipidemia,and people older than 50 years may increase the risk of KSHV infection.But the risk of coronary heart disease,high blood uric acid,hypoalbuminemia,sex,hypertension,malignant tumor,HIV,HBV,and occupation,level of education,marital status,if marry with infections such as the current status and monthly income situation KSHV are no obvious correlation or no statistical significance,it may not be a factor affecting the risk of infection. |
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