Detection of adeno-associated virus DNA in female genital samples by PCR-ELISA.
Adeno-associated viruses (AAV) are human parvoviruses that require helper function for their replication. Several studies have demonstrated that AAV DNA sequences can be found in the female genital tract but the incidence of infection seems very variable. A PCR-ELISA method detecting AAV DNA was developed for combining the specificity and the sensitivity of conventional PCR with an objective interpretation of the results. In the PCR-ELISA, a defined number of cells from cervical specimens were digested and amplified with concomitant digoxigenin labeling. Digoxigenin-labeled amplified products hybridized to a specific biotinylated probe were captured in streptavidin-coated microtiter wells by a biotin-streptavidin binding and were visualized by colorimetric immunoenzymatic reaction. PCR-ELISA was carried out in 110 cervical cytological specimens of women with or without the concomitant detectable presence of papillomavirus (HPV) DNA and the results were compared with those obtained by conventional PCR followed by dot blot hybridization. When compared to conventional PCR considered as reference standard, PCR-ELISA was found to be 98% sensitive and 96% specific. Out of the total 110 samples examined, 52.7% were positive for AAV DNA by both techniques, demonstrating a high prevalence of AAV infection in the uterine cervix. When analyzing samples with or without the presence of HPV DNA, 63.2 % of the samples were positive for HPV DNA and 41.5% of the samples were negative for HPV proved positive for AAV DNA by both PCR-ELISA and conventional PCR. Hence, PCR-ELISA, which can be completed in 1 day, proved to be a reliable method for an objective detection of AAV DNA in clinical samples. The present study showed a frequent infection of the cervical epithelium with AAV both in the presence and absence of HPV infection.
Venturoli
S
S
Division of Microbiology, Department of Clinical and Experimental Medicine, University of Bologna, Italy.
Cricca
M
M
Bonvicini
F
F
Gallinella
G
G
Gentilomi
G
G
Zerbini
M
M
Musiani
M
M
D002584:Cervix Uteri / Q000821:virology; D004279:DNA, Viral / Q000032:analysis*; D000229:Dependovirus / Q000235:genetics / Q000302:isolation & purification*; D004076:Digoxigenin; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D005836:Genitalia, Female / Q000821:virology*; D006801:Humans; D007558:Italy; D027383:Papillomaviridae; D030361:Papillomavirus Infections / Q000821:virology; D010322:Parvoviridae Infections / Q000821:virology*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D019809:Streptavidin; D014412:Tumor Virus Infections / Q000821:virology
D004279:DNA, Viral; D019809:Streptavidin; D004076:Digoxigenin