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Results for “Paraffin Embedding”

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PubMed2002

HOPE--a new fixing technique enables preservation and extraction of high molecular weight DNA and RNA of > 20 kb from paraffin-embedded tissues. Hepes-Glutamic acid buffer mediated Organic solvent Protection Effect.

The growing number of molecular pathologic tools that are currently available require material with good long term preservation of morphology, nucleic acids, and antigenic structures. However, pathologic investigations of tissues done at a molecular level are often hampered by the fixatives in use. We thus endeavored to design a new fixing system, including subsequent paraffin-embedding and sectioning, that makes complete pathologic analyses possible, with special consideration of immunohistochemistry (IHC), in situ hybridization (ISH), and molecular pathology. The optimized HOPE (Hepes-Glutamic acid buffer mediated Organic solvent Protection Effect) fixing technique allows us to preserve and extract high molecular weight DNA and RNA of > 20 kbp suitable for downstream applications, such as PCR and RT-PCR from HOPE-fixed, paraffin-embedded tissues that are up to 5 years old. This technique will most probably lead to new impacts on molecular pathology.

Cross-Linking Reagents↗

PubMed1994

Fiber-type differentiation by myosin immunohistochemistry on paraffin-embedded skeletal muscle. A useful adjunct to fiber typing by the adenosine triphosphatase reaction.

The myofibrillar adenosine triphosphatase reaction has been traditionally used to delineate fiber types in skeletal muscle biopsy specimens. At a pH of 9.4, the type 2 fibers are stained and the type 1 fibers remain unstained, thus permitting a clear distinction between the two fiber types and allowing morphometric assessments of fiber size, fiber diameter, and fiber ratios. However, this histochemical reaction requires frozen muscle tissue and is not feasible in situations when only paraffin-embedded tissue is available. We describe a useful alternative to determine fiber types in skeletal muscle specimens by myosin histochemistry on formalin- or Bouin-fixed, paraffin-embedded tissue. Our experience with over 300 muscle samples from both surgical and autopsy specimens indicates that myosin immunohistochemistry also provides accurate assessments of fiber types, fiber sizes, and fiber ratios. The most significant advantage of this method is its applicability to nonfrozen material, thus permitting fiber-type distinction in situations when frozen tissue is unavailable and facilitating retrospective analysis on archival material.

Humans↗

PubMed1980

A technique for localizing LDL by immunofluorescence in formalin-fixed and paraffin-embedded atherosclerotic lesions.

A protocol has been developed by which apo beta, the protein component of plasma low density lipoproteins (LDL), can be localized in four micron-thick sections of formalin-fixed and paraffin-embedded arteries with atherosclerotic involvement from man or non-human primate. An immunofluorescence procedure was employed that utilized affinity column-purified antibody coupled to fluorescein isothiocyanate (FITC). Although fixation reduced the intensity of fluorescence relative to that in cryostat sections, sufficient sensitivity was usually achieved when microscopy was performed using incident-light fluorescence together with a filter system adjusted for maximum absorption of FITC and by using oil immersion objectives. Furthermore, the use of various counterstains altered the color of the autofluorescence, normally strongly yellow, thus greatly enhancing the contrast and visualization of the apple green specific fluorescence. Non-chelated eriochrome black reduced tissue autofluorescence to barely visible levels, a feature useful in black and white photographs to demonstrate the localization pattern of LDL in arteries. This stain also reduced non-specific fluorescence. Other counterstains that proved useful were emthyl green, flazo orange and chelated eriochrome black. The latter two were particularly valuable for cellular detection. At low magnification LDL localization in atherosclerotic lesions from both human aortas and coronary arteries from monkeys fed an atherogenic diet, gave a pattern similar to those reported in cryostat sections of human lesions. At higher magnification, the improved detail of LDL localization could presumably be discerned as a result of immobilization of the soluble antigen by fixation, and by the ability to cut thinner sections of embedded material. This immunofluorescence procedure to localize LDL in formalin-fixed and paraffin-embedded atherosclerotic lesions should prove useful in both prospective as well as retrospective studies on clinical or chemical-pathological correlative studies on human tissues or in studies on atherogenesis using animal models.

Arteriosclerosis↗

PubMed2001

Cyclin D1 overexpression in multiple myeloma. A morphologic, immunohistochemical, and in situ hybridization study of 71 paraffin-embedded bone marrow biopsy specimens.

Cyclin D1 expression was evaluated by immunohistochemical analysis and biotin-labeled in situ hybridization (ISH) in a series of 71 decalcified, paraffin-embedded bone marrow biopsy specimens from patients with multiple myeloma (MM). Cyclin D1 messenger RNA (mRNA) overexpression was detected by ISH in 23 (32%) of 71 cases, whereas cyclin D1 protein was identified by immunohistochemical analysis in 17 (24%) of 71 specimens. All cases that were positive by immunohistochemical analysis also were positive by ISH. Statistically significant associations were found between cyclin D1 overexpression and grade of plasma cell differentiation and between cyclin D1 overexpression and extent of bone marrow infiltration. Our findings demonstrate the following: (1) ISH for cyclin D1 mRNA is a sensitive method for the evaluation of cyclin D1 overexpression in paraffin-embedded bone marrow biopsy specimens with MM. (2) ISH is more sensitive than immunohistochemical analysis in the assessment of cyclin D1 expression. (3) Cyclin D1 overexpression in MM is correlated positively with higher histologic grade and stage.

Antigens, CD20↗

PubMedDate not supplied

Steroid receptor heterogeneity in relation to DNA index in breast cancer: a multiparameter flow cytometric approach on paraffin-embedded tumor samples.

Steroid hormone (estrogen and progesterone) receptor (ER and PR) status at the time of breast carcinoma surgery is used as a marker for hormone dependency to guide adjuvant therapy. In a significant number of cases a discrepancy exists between the detected number of hormone receptors and the response to hormonal therapy. One of the explanations for this could be intratumoral heterogeneity. Our objective was to investigate the heterogeneity of steroid hormone receptor expression in breast cancer by using multiparameter flow cytometry (MP-FCM) on routinely processed formalin-fixed, paraffin-embedded tumors. A series of 232 routinely processed breast carcinomas were analyzed using a recently developed technique for the isolation of single cells from paraffin-embedded material. After dewaxing and rehydrating, 50-microm thick sections were heated for 2 hours at 80 degrees C in a citrate solution. Single-cell suspensions were prepared by a short pepsin digestion. The obtained single-cell suspensions were immunostained simultaneously for cytokeratin and ER or PR. Finally, DNA was stained using propidium iodide, after which the samples were analyzed on a flow cytometer. The fractions of ER- and PR-positive cells were determined in the total, as well as the G0 /G1 fraction of the diploid, and in case of nondiploid tumors, also in the G0 /G1 fraction of the aneuploid cell population. Of 232 cases, 88 (38%) were diploid, 38 (16%) were tetraploid, and 106 (46%) were aneuploid. In the diploid tumors the mean fraction of ER- and PR-positive cells was 81% and 76%, respectively. The ER- and PR-positive fractions in the total cytokeratin-positive fraction decreased significantly in the tetraploid (56% and 55%, respectively) and aneuploid tumors (both 47%, p < 0.0001). When analyzing the ER- and PR-positive fractions separately in the diploid and aneuploid cell populations of the nondiploid tumors, it became apparent that the ER and PR status in the diploid fraction of the tumor was significantly higher than in the aneuploid fraction (p < 0.0001). For the tetraploid tumors the mean ER- and PR-positive fractions were 79% and 76%, respectively, in the diploid fraction, and this decreased to 45% in the aneuploid cell subpopulation. In the aneuploid tumors this decrease was even more drastic: in the diploid cell population the ER- and PR-positive fractions were 66% and 62%, while this was 38% and 39% in the aneuploid population. These findings illustrate clearly the existence of a heterogeneous distribution of ER/PR expression in breast cancer, related to the loss of a diploid DNA index. Because of its objective quantification of subfractions within the same tumor, MP-FCM can be regarded as a superior method compared to more conventional techniques such as immunohistochemistry and biochemistry.

Adult↗

PubMed1999

An evaluation of immunofluorescence and PCR methods for detection of rabies in archival Carnoy-fixed, paraffin-embedded brain tissue.

Direct immunofluorescence and PCR detection methods were compared for sensitivity in evaluating the rabies status of archival specimens of Carnoy-fixed, paraffin-embedded brain tissue. The material consisted of 23 samples obtained during a rabies outbreak in Finland in 1988, and one sample isolated from a bat researcher who died of rabies in Finland in 1985. These results were compared with the original diagnoses performed on the fresh tissues. The immunofluorescence assay detected 100% (12/12) of the rabies-positive archival cases. A PCR assay designed to detect a 139-bp target near the 5' end of the rabies nucleoprotein gene also detected 100% (12/12) of the samples identified as positive in the fresh tissue specimens. A PCR assay designed to detect a 304-bp target spanning the 139-bp target of the first assay detected only 67% (8/12) of the original cases. No false positives were recorded. Both immunofluorescence detection of antigen and PCR detection of a short region of the nucleoprotein gene are useful in determining the rabies status of fixed, paraffin embedded (archival) material.

Animals↗

PubMed1995

Specific immunohistochemical identification of Candida albicans in paraffin-embedded tissue with a new monoclonal antibody (1B12).

In invasive candidiasis, the identification of Candida organisms in tissue samples or in normally sterile fluids is essential for an accurate diagnosis. Species identification is an important clue for the source of infection and in epidemiological studies. In this article, the authors have tested the value of a new monoclonal antibody (1B12) to detect C albicans in culture by immunofluorescence, and in tissue samples by immunohistochemistry. MAb 1B12 was found to specifically recognize C albicans, does not cross-react with other Candida species or other structurally similar fungi, and is very sensitive and specific in paraffin-embedded tissue, having no reactivity in normal human tissues or necrotic areas. Therefore, MAb may be a valuable tool in the evaluation of fungal infections in paraffin-embedded tissue, particularly when Candida species identification is needed.

Antibodies, Monoclonal↗

PubMed1989

Immunohistochemical staining properties of amyloids with anti-keratin antibodies using formalin-fixed, paraffin-embedded sections.

Immunohistochemical staining properties of amyloids with anti-keratin antibodies were investigated using an avidin-biotin-peroxidase complex (ABC) system on formalin-fixed, paraffin-embedded sections. Anti-keratin antibody EAB-903 which recognize 66K and 57K daltons keratin peptides reacted with amyloid deposits in both lichen amyloidosus (LA) and macular amyloidosis (MA), but did not react with either primary systemic amyloidosis (AL), secondary systemic amyloidosis (AA) or heredofamilial amyloid polyneuropathy (AF). However, anti-keratin antibodies EAB-904 and MAK-6 did not react with any types of amyloids. These results suggested that immunohistochemical staining with anti-keratin antibody EAB-903 using formalin-fixed, paraffin-embedded sections appeared to be a useful method in making differential diagnosis of primary localized cutaneous amyloidosis (AD).

Amyloid↗

PubMed1994

Intravascular (angiotropic) large cell lymphoma: determination of monoclonality by polymerase chain reaction on paraffin-embedded tissues.

Angiotropic lymphoma is a rare, aggressive, intravascular non-Hodgkin's lymphoma, usually of B-cell phenotype. Because lymphoma is often clinically unsuspected, the small skin or muscle biopsies typically obtained for evaluation make assessment of lymphoid clonality through cell surface markers or Southern blot hybridization analysis difficult or impossible. The recent development of polymerase chain reaction methodologies to detect chromosomal translocations and immunoglobulin heavy chain gene rearrangement on paraffin-embedded tissue offers an attractive alternative for ascertaining the clonality of lymphoproliferative processes. We report a case of B-cell angiotropic lymphoma in which a monoclonal variable diversity joining region rearrangement of the immunoglobulin heavy chain locus was detected by polymerase chain reaction in both ante- and postmortem, formalin-fixed, paraffin-embedded skeletal muscle. The use of polymerase chain reaction in assessing clonality in angiotropic lymphoma is enhanced by the general absence of a background of reactive B-lymphoid cells in angiotropic lymphoma, which can obscure the monoclonal band and/or compromise sensitivity. No amplification product was obtained for t(14;18) involving the bcl-2 major breakpoint region. It is interesting to note that this case exhibited rare circulating lymphoma cells and more extensive bone marrow involvement (more than 100 tumor cells/high magnification field) than has been previously described.

Aged↗

PubMed1998

A new Mel-CAM (CD146)-specific monoclonal antibody, MN-4, on paraffin-embedded tissue.

Mel-CAM (also termed CD146 or MUC18) is a cell-adhesion molecule belonging to the immunoglobulin gene superfamily. It mediates cell-cell interaction through heterophilic Mel-CAM/ligand adhesion. Previous studies showed Mel-CAM immunoreactivity in normal and neoplastic human tissues, but an extensive assessment of Mel-CAM distribution was not performed because of the lack of a Mel-CAM-specific monoclonal antibody that could be used in routinely processed, formalin-fixed, paraffin-embedded tissues. Therefore, we developed a mouse monoclonal antibody, MN-4, that specifically recognized a fixation-resistant epitope in the second extracellular (C2) domain of Mel-CAM. We performed immunohistochemical staining on 467 paraffin-embedded tissue samples with this MN-4 antibody and avidin-biotin peroxidase. MN-4 immunoreactivity was seen in normal tissues, including endothelium, smooth muscle, epithelial and myoepithelial cells of the breast, Schwann cells, ganglion cells, cerebellar cortex, implantation-site intermediate trophoblast, external root sheath of hair follicles, basal cells of bronchial epithelium, parathyroid glands, subcapsular epithelium of thymus, follicular dendritic reticulum cells, skeletal muscles, epithelium of lens, and glial cell fibers in the central nervous system in early embryos. In malignant tumors, MN-4 immunostaining was consistently present in all melanomas, angiosarcomas, Kaposi's sarcomas, leiomyosarcomas, mucoepidermoid carcinomas of salivary gland, placental-site trophoblastic tumors, and choriocarcinomas. Three of 8 squamous cell carcinomas, 2 of 2 small cell carcinomas of the lung, 2 of 11 of infiltrating breast carcinomas, and 4 of 11 of germinomas were focally and weakly positive for MN-4 antibody. In contrast, a wide variety of other carcinomas, sarcomas, lymphomas, leukemias, and neuroendocrine tumors failed to show MN-4 immunoreactivity. In conclusion, MN-4 is a specific monoclonal antibody that recognizes Mel-CAM on paraffin sections. This antibody is useful in retrospective studies of Mel-CAM expression in archival tissue sections and might provide a diagnostic and prognostic marker for human neoplasms.

Animals↗

PubMed1999

Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR/DGGE)-based detection of clonal T-cell receptor gamma gene rearrangements in paraffin-embedded cutaneous biopsies in cutaneous T-cell lymphoproliferative diseases.

Polymerase chain reaction (PCR)-based amplification of T-cell receptor (TCR)-gamma genes is a novel technique that can detect a clone of T cells comprising less than 1% of the total T cells in a lymphoid infiltrate. Besides greater sensitivity than Southern blotting, this technique can be performed with smaller quantities of lower molecular weight genomic DNA as template. We retrospectively analyzed 12 paraffin-embedded biopsies of cutaneous T-cell lymphoma (CTCL), 1 case suspicious for CTCL, 1 case of granulomatous slack skin, and 8 cases of inflammatory skin diseases to determine if PCR-denaturing gradient gel electrophoresis (PCR-DGGE) analysis can detect TCR-gamma gene rearrangements on paraffin-embedded specimens. We were able to amplify Vgamma1-8 TCR sequences in each case and detected a dominant clone in 9 of 12 cases of CTCL and in granulomatous slack skin. We analyzed Vgamma9 sequences in 9 cases of CTCL and detected a dominant clone in 4 cases. This study demonstrates that PCR-DGGE can easily be applied retrospectively to cutaneous biopsies of lymphoproliferative diseases when fresh tissue is not available.

Adolescent↗

PubMed1995

Detection of HCV-RNA in paraffin-embedded liver biopsies from patients with autoimmune hepatitis.

BACKGROUND/AIMS: There may be a relationship between autoimmune hepatitis and viral infection. To examine this relationship, 19 patients with autoimmune hepatitis and/or chronic hepatitis C were studied. METHODS: Patients were selected initially on the basis of having autoantibodies (anti-nuclear, anti-smooth muscle, or anti-liver-kidney microsomal) in serum. Formalin-fixed, paraffin-embedded liver biopsies from these patients were tested for HCV-RNA by polymerase chain reaction. The biopsies were examined histologically to detect features suggestive of chronic hepatitis C or autoimmune hepatitis. The results were correlated with serum anti-HCV and HCV-RNA, and with response to steroid therapy. RESULTS: Five of the nineteen patients had detectable HCV-RNA in their liver biopsies. In two of three patients from whom serum was available, HCV-RNA was detectable. The remaining 14 patients were negative for HCV-RNA by tissue polymerase chain reaction. Serum was available from 11 of these patients, and serum HCV-RNA was negative in all. All of the three HCV-RNA-positive patients who were treated with steroids showed a partial response; tissue positivity for HCV-RNA was significantly higher in partial responders than in complete responders (60% vs 0%, p = 0.01). Severe portal and periportal inflammation with prominent plasma cells together with bridging parenchymal necrosis were seen more often in HCV-negative biopsies. Mild portal and periportal inflammation with portal lymphoid aggregates, apoptosis and spotty parenchymal necrosis were seen more in HCV-positive biopsies. CONCLUSIONS: These results show that hepatitis C virus can be detected in some patients with circulating autoantibodies. The ability to detect HCV-RNA in paraffin-embedded archival material provides a valuable addition to the battery of available HCV tests.

Adolescent↗

PubMed1994

Analysis of changes in DNA sequence copy number by comparative genomic hybridization in archival paraffin-embedded tumor samples.

Analysis of previously unknown genetic aberrations in solid tumors has become possible through the use of comparative genomic hybridization (CGH), which is based on competitive binding of tumor and control DNA to normal metaphase chromosomes. CGH allows detection of DNA sequence copy number changes (deletions, gains, and amplifications) on a genome-wide scale in a single hybridization. We describe here an improved CGH technique, which enables reliable detection of copy number changes in archival formalin-fixed paraffin-embedded tumor samples. The technique includes a modified DNA extraction protocol, which produces high molecular weight DNA which is necessary for high quality CGH. The DNA extraction includes a 3-day digestion with proteinase K, which remarkably improves the yield of high molecular weight DNA. Labeling of the test DNA with a directly fluorescein-conjugated nucleotide (instead of biotin labeling) improved significantly the quality of hybridization. Using the paraffin-block technique, we could analyze 70 to 90% of paraffin blocks, including very old samples as well as samples taken at autopsy. CGH from paraffin blocks was highly concordant (95%) with analyses done from matched freshly frozen tumor samples (n = 5 sample pairs; kappa coefficient = 0.83). The method described here has wide applicability in tumor pathology, allowing large retrospective prognostic studies of genetic aberrations as well as studies on genetic pathogenesis of solid tumors, inasmuch as premalignant lesions and primary and metastatic tumors can be analyzed by using archival paraffin-embedded samples.

Chromosome Mapping↗

PubMed1996

Retrospective survey of chronic Q fever in Japan by using PCR to detect Coxiella burnetii DNA in paraffin-embedded clinical samples.

We used PCR to detect Coxiella burnetii DNA in paraffin-embedded tissues obtained from patients with chronic endocarditis in which the etiological agent had been unknown. On the basis of the published nucleotide sequence of the C. burnetii htpB gene, primers were chosen to produce an amplified fragment of 285 bp. A total of 60 samples from 56 patients were tested for the presence of C. burnetii DNA. Five samples from four patients were found to be positive. All of the amplified DNA fragments possessed a TthHB8I restriction site, as predicted from the published sequence of C. burnetii. In one of the four positive patients, rickettsia-like particles were found in sections of tissue stained by Gimenez's method. This is the first report of chronic Q fever in Japan.

Base Sequence↗

PubMed1990

New technique for processing paraffin-embedded tissue for Y chromosome fluorescence identification of trophoblastic disease.

A new reprocessing technique for Y chromosome fluorescent body (q 12 region) detection of trophoblastic disease in previously paraffin-embedded tissues is described. Deparaffinized sections were treated with pronase and trypsin for digestion, followed by hydrolysis with HCl and acetic acid, staining with quinacrine hydrochloride fluorochrome and mounting in Sørensen's phosphate buffer (pH 5.5). Use of the technique resulted in sufficient fluorescence quality and better accuracy for Y and X heterochromatin scoring. The technique yielded the same results in retrospective formalin-fixed, paraffin-embedded trophoblastic specimens as in fresh tissues. The combinations of enzymes and acids and the dosages necessary for optimal results are discussed.

Female↗

PubMed2005

Comparison of archival plasma and formalin-fixed paraffin-embedded tissue for genotyping in hepatocellular carcinoma.

Biobanks containing formalin-fixed paraffin-embedded tissue, as well as frozen serum or plasma, are important resources for molecular epidemiologic studies. However, few studies have compared the reliability of formalin-fixed tissue samples and archival plasma samples for genotyping. We determined the genotype of four proposed genetic risk factors for hepatocellular carcinoma [hereditary hemochromatosis (HFE 63 and 282), alpha(1)-antitrypsin deficiency (AAT 342) and cystic fibrosis (CFTR 508)] on formalin-fixed tissue samples, stored for up to 25 years, from 318 patients diagnosed with hepatocellular carcinoma and on plasma or serum samples from 31 of these patients. The genotypes were analyzed by RFLP or allele-specific amplification as well as by TaqMan assays. In addition, genotyping was attempted after whole genome amplification by multiple displacement amplification (MDA). Genotyping was successful in 94% of the tissue samples and successful and identical to the tissue samples from the same subjects in 98% of the plasma/serum samples. DNA from plasma samples could be amplified >5,000-fold by MDA and genotyping after MDA gave identical results to the genotyping of the same subjects before whole genome amplification. MDA amplification of the tissue samples was not successful. In summary, archival plasma was found to be an adequate source of efficiently amplifiable DNA. MDA on plasma samples allows analysis of multiple genotypes in epidemiologic studies.

Carcinoma, Hepatocellular↗

PubMed1999

Detection of Mycobacterium avium subspecies avium in formalin-fixed, paraffin-embedded tissues of captive exotic birds using polymerase chain reaction.

A presumptive diagnosis of avian tuberculosis can be made when characteristic histologic lesions and acid-fast bacilli are observed in avian tissue samples. However, a definitive diagnosis requires isolation and identification of the causative organism, a process that can take several weeks to complete. The purpose of the study was to determine whether formalin-fixed, paraffin-embedded archival avian tissues could be tested by polymerase chain reaction (PCR) to reliably and rapidly diagnose avian tuberculosis. Tissues were examined from both presumptive and definitive cases of avian tuberculosis from captive exotic birds obtained over a 14-yr period (1983-1997). The cases chosen consisted of birds that had characteristic histologic lesions with acid-fast bacilli. The primers used for PCR amplified a 180-base-pair fragment of 16S ribosomal RNA, a sequence specific for both Mycobacterium avium subsp. avium and M. avium subsp. paratuberculosis. If a sequence was detected in a sample, it was presumed that M. a. avium was the organism being detected. This M. avium fragment sequence was detected in 26 of the 97 samples (27%). Some of the negative PCR results may be explained by any of several factors that adversely affect nucleic acid integrity, particularly prolonged fixation in formalin. Of the 17 samples that were culture positive for M. avium and were known to have been fixed in formalin for < or = 4 wk, 11 tested positive by PCR (65%). The findings of this study show that PCR can be a rapid indicator of the presence of M. a. avium in formalin-fixed, paraffin-embedded tissues. However, the relatively low detection rate the test demonstrated in this sample set may limit its practical use as a diagnostic tool.

Air Sacs↗

PubMed1992

Human papillomavirus DNA in cervix. In-situ hybridization with biotinylated probes on Bouin's fixed paraffin embedded specimens.

We examined retrospectively a series of 65 Bouin's fixed, paraffin-embedded tissue specimens from 8 condylomatous lesions, 16 condylomas associated with cervical intraepithelial neoplasia (CIN), and 12 neoplasia without condylomatous signs, for histological characteristics, the detection of viral structural antigen, the presence and typing of HPV DNA by molecular in situ hybridization with biotinylated probes types 6, 11, 16 and 18 under stringent conditions (Tm - 12 degrees C). HPV DNA was present in 34/65 (52%) specimens. Detection of viral structural antigen was positive in only 14% (3/22) specimens. HPV DNA were identified in 9/9 (100%) condylomatous lesions (with HPV type 6, 11, 18). Three condylomas were coinfected with both HPV type 6 or 11 and type 18; viral antigen was found in two specimens. HPV DNA were detected in 18/31 (58%) low grade and advanced CIN associated with condylomatous changes (type 6 = 5 specimens, type 11 = 3 specimens, type 16 = 4 specimens, type 18 = 6 specimens). Four of these cases were coinfected with both HPV type 6/11 and HPV type 16/18. Viral antigen was negative in all specimens. HPV DNA were detected in 7/25 (28%) advanced intra-cervical neoplasia (CIN III) without anatomopathological condylomatous changes (type 6 = 1 specimen, type 16 = 3 specimens, type 18 = 3 specimens). One of these specimens contained both HPV types 6 and 18. Viral antigen was found in one case. Our data confirm the association of HPV types 6 and 11 with condyloma and low grade neoplasia; HPV types 16 and 18 were associated with advanced cervical neoplasia.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetates↗