| BackgoundCondyloma acuminatum (CA) is a common and easy relapsed sexually transmitted disease caused by human papilloma virus (HPV), which dose harm to the patient’s somatopsychic health, reproductive health and family life. Among them, more than 90% genital warts caused by HPV6 and 11. Recently, the incidence of CA is rising constantly, and the majority of these are subclinical and latent infection, which may affect the prevalence, transmission, duration and prognosis of CA. However, lack of convenient and accurate method in detecion of condyloma acuminatum, especially in subclinical and latent infection, misdiagnosis and missed diagnosis always occur. The methods of HPV pathogen detection, such as nucleic acid molecular technology, for its high quantity of laboratory conditions, can not be widely used in clinical work especially in primary hospitals. In addition, for the low detection rate of the L1 protein of HPV, immunohistochemical detection, though it has a simple operation process, also could not be widely used in clinic. In this study, the significance and clinical applicabillity of immunohistochemical detection in CA tissue with home-made anti-HPV6b/11 E7 antibody will be evaluated.ObjectiveTo detect the expression of HPV6b and 11 E7 protein on condyloma acuminatum lesions of vulva with home-made anti-HPV6b/11 E7 antibody. To observe the coincidence rate with clinical diagnosis and the results of RT-PCR, and evaluate the clinical application of these two antibodies.Methods1. Total 55 condyloma acuminatum samples were collected in the dermatological department of the First Hospital of Fuyang Hangzhou from Sepetember 2013 to October 2014. After routine disinfection, the condyloma acuminatum tissues were taken with the blood vessel forcep under local anesthesia. The tissues were fixed with formol after normal saline irrigation, which will be checked by routine pathological microscopy.18 samples were cryopreserved at -80℃.2.55 cases of condyloma acuminatum paraffin-embedded tissues were detected by immunohistochemistry staining with anti-HPV 6b/11 E7 protein polyclonal antibody. The cell dyeing were observed by microscope. In positive staining cases, the location, intensity and percentage of positive cells were analysed.3. The mRNA expression of HPV6b and 11 E7 protein of 18 cases were detected by RT-PCR, and the coincidence rate of both detection methods were analysed.ResultsBy immunohistochemical stain, we found the expression of HPV6b andll E7 protein was all in the cell nuclear of full layer epidermis and more in the basal cell. HPV6b E7 protein was expressed in 42 cases (positive rate 76.36%), while HPV11 E7 protein was expressed in 32 cases (positive rate 58.18%). HPV6b and 11 E7 positive cases were detected in 52 cases (HPV6b E7 plus HPV11 E7 positive rate 94.55%), of which, both HPV6b and 11 E7 positive were detected in 22 cases (double positive rate 40.00%). Only 3 cases were detected negative for both two HPVs E7 (negative rate 5.45%). Among 42 HPV6b E7 positive cases,20 cases were detected negative for HPV11 E7; among 32 HPV11 E7 positive cases,10 cases were detected negative for HPV6b E7. By RT-PCR, we found the HPV6b and 11 E7 mRNA expression were positive for 15 cases and 10 cases respectively. Both HPV6b and 11E7 mRNA expression was positive in 7 cases, which consists with the result of immunohistochemistry.DiscussionWe detected 55 cases of CA specimens with home-made anti-HPV 6b/11 E7 protein polyclonal antibody, and the total positive rate of HPV6 and 11 infection was 94.55%, which is in accordance with reported studies:more than 90% CA were infected with low-risk HPV 6 or HPV11. It is observed intuitively that HPV 6b/11 E7 positive expression cells dispersedly distributed in all layers of epidermis, and the expression of HPV6b and 11 E7 protein was all in the cell nuclear. This study suggested that the positive rate of home-made anti-HPV 6b/11 E7 protein polyclonal antibody detecting in CA tissues is high proficiency. In HPV6b E7 protein positive staining cases, only Some are positive expression of HPV 11 E7 too, and vice versa. The result demonstrated that there was no cross reaction between the two antibodies, and RT-PCR detection of 18 cases found that, the mRNA expression of HPV6b or HPV 11 was in accordance with the immunohistochemical detection completely, which meaned that both the HPV6b and 11 E7 antibody had high specificity.ConclusionsThe home-made anti-HPV6b/11E7 protein polyclonal antibodies are highly specific and sensitive for detecting HPV infection in condyloma acuminatum. There was no cross reaction between the two antibodies, and the HPV6b/11 virus infected cells can be observed intuitively in CA tissues. Immunohistochemical detection with these two home-made polyclonal antibodies has a simple operation process, so it would be helpful for clinical diagnoses of condyloma acuminatum and be worth to popularize in clinic. |
