ABSTRACT
AIMS: To evaluate the value of polymerase chain reaction-in situ hybridization (PCR-ISH) for the detection of human papillomaviruses (HPV) in paraffin sections of cervical biopsies fixed either in 10% formalin or in Bouin's solution. METHODS AND RESULTS: We analysed 40 biopsies from Italian women infected with the human immunodeficiency virus type 1 (HIV 1). In-situ hybridization techniques were performed with commercial biotinylated probes. The PCR-ISH was carried out by the 'hot start modification'. Cervical intraepithelial neoplasia (CIN) was found in 23 of 40 patients (57. 5%); eight cases showed condylomatous features. Human papillomavirus was detected in 42.5% by ISH and in 65% by PCR-ISH. Sixty-nine per cent of positive biopsies contained HPV 16, 18, 31 and 33. HPV 6 and 11 were found only in condylomata acuminata samples. CONCLUSIONS: The results point to a high incidence of HPV infection as well as of CIN in HIV-positive patients. Human papillomavirus type 16 appears to be most frequently associated with CIN. Polymerase chain reaction-ISH is more sensitive than ISH in the detection and typing of HPV DNA both in clinical and in 'latent' infections. The two techniques yielded the same results with either formalin- or Bouin's-fixed material.
Subject(s)
DNA, Viral/analysis , In Situ Hybridization/methods , Papillomaviridae/genetics , Papillomavirus Infections/diagnosis , Polymerase Chain Reaction/methods , Tissue Fixation/methods , Tumor Virus Infections/diagnosis , Acetic Acid , Adult , Condylomata Acuminata/diagnosis , Condylomata Acuminata/virology , Female , Fixatives , Formaldehyde , HIV Infections/complications , HIV-1 , Humans , Picrates , Uterine Cervical Neoplasms/diagnosis , Uterine Cervical Neoplasms/virology , Uterine Cervical Dysplasia/diagnosis , Uterine Cervical Dysplasia/virologyABSTRACT
The viral typization in Human Papillomavirus (HPV) infections of the lower female genital tract is relevant both from the epidemiological and the clinical point of view. We have tested DNA from specimens obtained by guided biopsies on neoplastic and benign lesions, using single probes of the different virus types (6, 11, 16 and 18) by Dot-blot and Southern blot analysis. According to previous studies HPV 16 and 18 have been detected in neoplastic lesions, while 6 and 11 were more frequently found associated to condylomata. Negative specimens and intermediate were 36% of the total. In our experience both methods have shown same sensibility.
Subject(s)
Papillomaviridae/classification , Tumor Virus Infections/microbiology , Uterine Cervical Neoplasms/microbiology , Vulvar Neoplasms/microbiology , Adenocarcinoma/microbiology , Adolescent , Adult , Aged , Carcinoma in Situ/microbiology , Carcinoma, Squamous Cell/microbiology , Condylomata Acuminata/microbiology , DNA Probes, HPV , DNA, Neoplasm/analysis , Female , Humans , Middle Aged , Nucleic Acid HybridizationABSTRACT
Sera of 1023 subjects belonging to categories at risk for AIDS, positive for HIV-1, were tested for antibodies to human T-cell leukemia virus (HTLV-1) by ELISA, Western blotting, and radioimmunoprecipitation (RIP) assay. Sera of these subjects were positive for antibodies to HTLV-1 in a percentage of 18.54%. Nine out of 18 sera from patients with AIDS had antibodies to HTLV-1 (50%). Sera of 207 intravenous drug abusers and 64 homosexual males, seronegative for HIV-1, were tested for antibodies to HTLV-1 and 6.7% of intravenous drug abusers and 7.8% of homosexual males were found to be HTLV-1 antibody positive, showing that HTLV-1 infection occurred independently of HIV-1 and HIV-2 infections. In fact the same HIV-1 seronegative patients were also tested for the presence of antibodies to HIV-2 and none of them was found to be positive.