Prevalence and determinants of high-risk human papillomavirus (HPV) infection and cervical cytological abnormalities in imprisoned women.

The aim of the study was to estimate the prevalence and risk factors associated with infection by high-risk human papillomavirus (HR-HPV) in cervix and squamous intra-epithelial lesions (SIL) in imprisoned women. This was done by a cross-sectional study of imprisoned women attending the gynaecological clinic in Foncalent prison in Alicante, Spain. The study period was from May 2003 to December 2005. HR-HPV infection was determined through Digene HPV Test, Hybrid Capture II (HC-II). HPV typing was determined by multiplex nested PCR assay combining degenerate E6/E7 consensus primers. Multiple logistic regression modelling was used for the analysis of associations between variables where some were considered possible confounders after checking for interactions. A total of 219 women were studied. HR-HPV prevalence was 27.4% and prevalence of SIL was 13.3%. HIV prevalence was 18%, higher in Spaniards than in migrant women (24.6% vs. 14.3%, P<0.05). In multivariate analyses, risk factors for HPV infection were younger age (P for trend=0.001) and tobacco use (OR 2.62, 95% CI 1.01-6.73). HPV infection (OR 4.8, 95% CI 1.7-13.8) and HIV infection were associated with SIL (OR 4.8, 95% CI 1.6-14.1). The commonest HPV types were HPV16 (29.4%), HPV18 (17.6%), HPV39 (17.6%) and HPV68 (17.6%). The prevalence of both HR-HPV infection and SIL in imprisoned women found in this study is high. Determinants for each of the outcomes studied were different. HPV infection is the most important determinant for SIL. A strong effect of HIV co-infection on the prevalence of SIL has been detected. Our findings reinforce the need to support gynaecological clinics in the prison setting.

Higher prevalence of human papillomavirus infection in migrant women from Latin America in Spain.

OBJECTIVES: To estimate prevalence and determinants of high risk (HR) human papillomavirus (HPV) by country of origin in women attending a family planning centre (FPC) in Alicante, Spain. METHODS: Cross sectional study of all women attending a FPC from May 2003 to January 2004. An ad hoc questionnaire was designed and data were collected prospectively. HR HPV infection was determined through the Digene HPV test, Hybrid Capture II, and positive samples for PCR were directly sequenced. Data were analysed through multiple logistic regression. RESULTS: HR HPV prevalence in 1011 women was 10% (95% CI: 8.2 to 12). Compared to Spaniards (prevalence 8.2%) HR HPV prevalence in Colombians was 27.5% (OR: 4.24 95% CI: 2.03 to 8.86), 23.1% in Ecuadoreans (OR: 3.35 95% CI: 1.30 to 8.63), and 22.73% in women from other Latin American countries (OR: 3.29 95% CI: 1.17 to 9.19). Women with more than three lifetime sexual partners had an increased risk of HR HPV infection (OR 3.21 95% CI: 2.02 to 5.10). The higher risk of HR HPV infection was maintained in Latin American women in multivariate analyses that adjusted for age, number of lifetime sexual partners, and reason for consultation. The commonest HPV types in women with normal cervical smears were HPV-18 (20%), HPV-16 (14%) and HPV-33 (11%). CONCLUSIONS: Prevalence of HR HPV is more than three times higher in Latin Americans than in Spaniards. Latin American women's HPV prevalence resembles more that of their countries of origin. It is essential that health service providers identify these women as a priority group in current cervical screening programmes.

Oncogenic human papillomavirus (HPV) type distribution and HPV type 16 E6 variants in two Spanish population groups with different levels of HPV infection risk.

The aim of this study is to determine oncogenic human papillomavirus (HPV) types and HPV type 16 (HPV16) variant distribution in two Spanish population groups, commercial sex workers and imprisoned women (CSW/IPW) and the general population. A multicenter cross-sectional study of 1,889 women from five clinical settings in two Spanish cities was conducted from May to November 2004. Oncogenic HPV infection was tested by an Hybrid Capture II (HC2) test, and positive samples were genotyped by direct sequencing using three different primer sets in L1 (MY09/11 and GP5+/GP6+) and E6/E7. HPV16 variants were identified by sequencing the E6, E2, and L1 regions. Four hundred twenty-five samples were positive for the HC2 test, 31.5% from CSW/IPW and 10.7% from the general population. HPV16 was the most frequent type. Distinct profiles of oncogenic HPV type prevalence were observed across the two populations. In order of decreasing frequency, HPV types 16, 31, 58, 66, 56, and 18 were most frequent in CSW/IPW women, and types 16, 31, 52, 68, 51, and 53 were most frequent in the general population. We analyzed HPV16 intratype variants, and a large majority (78.7%) belonged to the European lineage. AA variants were detected in 16.0% of cases. African variants belonging to classes Af1 (4.0%) and Af2 (1.3%) were detected. Different HPV types and HPV16 intratype variants are involved in oncogenic HPV infections in our population. These results suggest that HPV type distribution differs in CSW/IPW women and in the general population, although further analysis is necessary.

Analysis and management of HIV peptide microarray experiments.

OBJECTIVES: To develop a tool for then easy and user-friendly management of peptide microarray experiments and for the use of the results of these experiments for the study the immune response against HIV virus infection in clinical samples. METHODS: Applying bioinformatics and statistics for the analysis of data coming from microarray experiments as well as implementing a MIAME (Minimum Information About a Microarray Experiment) compliant database for managing and annotating experiments, results and samples. RESULTS: We present a new tool for managing not only nucleic acid microarray experiments but also protein microarray experiments. From the analysis of experimental data, we can detect different profiles in the reactivity of the sera with different genotypes. CONCLUSIONS: We have developed a new tool for managing microarray data including clinical annotations for the samples as well as the capability of annotating other microarray formats different to those based on nucleic acids. The use of peptide microarrays and bioinformatics analysis opens a new scope for the characterization of the immune response, and analyzing and identifying the humoral response of viruses with different genotypes.

Influence of age and geographical origin in the prevalence of high risk human papillomavirus in migrant female sex workers in Spain.

OBJECTIVES: To estimate the prevalence and risk factors of high risk human papillomavirus (HPV) infection in migrant female sex workers (FSW) according to age and geographical origin. METHODS: Cross sectional study of migrant FSW attending a sexually transmitted infection (STI) clinic in Madrid during 2002. Information on sociodemographic characteristics, reproductive and sexual health, smoking, time in commercial sex work, history of STIs, HIV, hepatitis B, hepatitis C, syphilis, and genitourinary infections was collected. High risk HPV Infection was determined through the Digene HPV Test, Hybrid Capture II. Data were analysed through multiple logistic regression. RESULTS: 734 women were studied. Overall HPV prevalence was 39%; 61% in eastern Europeans, 42% in Ecuadorians, 39% in Colombians, 29% in sub-Saharan Africans, and 24% in Caribbeans (p = 0.057). HPV prevalence showed a decreasing trend by age; 49% under 20 years, 35% in 21-25 years,14% over 36 years% (p<0.005). In multivariate analyses, area of origin (p = 0.07), hormonal contraception in women not using condoms (OR 19.45 95% CI: 2.45 to 154.27), smoking, age, and an interaction between these last two variables (p = 0.039) had statistically significant associations with HPV prevalence. STI prevalence was 11% and was not related to age or geographical origin. CONCLUSIONS: High risk HPV prevalence in migrant FSW is elevated and related to age, area of origin, and use of oral contraceptives in women not using condoms. These data support the role of acquired immunity in the epidemiology of HPV infection and identifies migrant FSW as a priority group for sexual health promotion.

HIV type 1 non-B subtype prevalence in Spain, 1997-1998.

We have evaluated the prevalence of HIV-1 non-B subtypes in Spain by means of an enzyme immunoassay (EIA) for discrimination between B and non-B subtypes. Samples were obtained from newly diagnosed patients attended at internal medicine outpatient clinics between October 1997 and October 1998. Discrimination between HIV-1 B and non-B subtypes was carried out by means of the EIA, with V3 synthetic peptides specific to the different subtypes. Non-B-serotyped samples were genetically analyzed in the gp41 region from the original sera. During the study period, 909 samples were collected from 21 medical units located in various Spanish geographical regions. Serotyping was possible in 885 cases, of which 791 were assigned as B serotype (89.38%), 70 showed no reactivity to any of the peptides (7.91%), and the remaining samples displayed other reaction patterns (2.72%). Of the 94 non-B-assigned samples, 65 were genetically characterized in the gp41 region of the env gene: 55 were B subtype, 5 were A subtype (4 clustered with CRF02AG reference strains), 3 were C subtype, and 2 were G subtype. The prevalence rate for non-B subtypes in Spain was established at 1.13% (95% CI, 0.59-2.21). Although the B subtype is predominant in the Spanish population, other subtypes have been detected.

Seroprevalence of HIV and HTLV in a representative sample of the Spanish population.

HIV and HTLV seroprevalence was determined by means of unlinked anonymous testing of 2144 sera, originally obtained from primary care patients by representative sampling of the Spanish population aged 15-39 years in 1996. HIV-1 seroprevalence was 4.3 per 1000 population in the 15-39 years age group [95% confidence interval (CI), 1.5-10.7] and 5.6 per 1000 (95% CI, 1.8-15.3) in the 20-39 years age group. Seroprevalence proved higher in males and urban residents. No antibodies to HIV-2 and HTLV-I were detected in any of the sera studied. However, presence of antibodies to HTLV-II was confirmed in one serum sample, while HTLV seroreactivity, though detected in another, could not be typed. The two HTLV-positive results equated to a seroprevalence of 1.9 per 1000 in the 20-39 years age group (95% CI, 0.3-8.6). HIV-I seroprevalence was consistent with previous estimates yielded by back-calculation. The level of HTLV seroprevalence found suggests endemicity.

Nucleotide sequence and restriction fragment-length polymorphism analysis of human T-cell lymphotropic virus type II (HTLV-II) in southern Europe: evidence for the HTLV-IIa and HTLV-IIb subtypes.

Human T-cell lymphotropic virus type II (HTLV-II) has been subtyped into two major groups, IIa and IIb, according to molecular studies involving env gene sequencing. Subsequently, this retrovirus was further subclassified by examining the long terminal repeat (LTR), the most divergent genomic region. Sequence analysis and restriction fragment-length polymorphism (RFLP) applied to the LTR region identified either four or five groups within the IIa subtype (depending on the restriction enzyme sets used) and six within the IIb subtype. In this study, we analyzed the LTR sequences of 29 samples obtained from HTLV-II-infected individuals living in Spain and Italy, which included 24 injecting drug users (IDUs), three blood donors, and two subjects at risk for HIV/HTLV infection. Sequence analysis and phylogenetic analysis of 720 base pairs of the LTR performed in 10 Spanish samples showed that all of these samples belonged to IIb subtype, with a divergence of 7.5% and 1.66% compared with MoT (IIa) and NRA/G12 (IIb) isolates, respectively. RFLP analysis demonstrated the presence of the IIb 4-subtype restriction pattern in 26 samples, a IIb5-subtype pattern in one Italian IDU, and a IIa0-subtype pattern in two Italian samples (blood donors), according to W.M. Switzer's nomenclature. This is the first report of the presence of IIb5 in Southern Europe and IIa0 among Italian blood donors. RFLP correlated with nucleotide sequence and phylogenetic data obtained in this study, demonstrating the ability of the RFLP method to predict the phylogroup of HTLV-II-infected samples.

Typing human T-cell lymphotropic virus (HTLV-I and HTLV-II) by nested polymerase chain reaction: application to clinical specimens.

Human T-cell lymphotropic virus type I and II provirus DNA was detected by polymerase chain reaction (PCR). MT-2 (HTLV-I infected), C3/44 Mo (HTLV-II infected) cell lines and peripheral blood mononuclear cells (PBMNC) from HTLV seropositive samples were used. The procedure consists of first amplification which detects both HTLV-I and HTLV-II, and a second amplification (nested-PCR) to discriminate between the two viruses and to improve sensitivity. Optimal conditions of MgCl2 concentration and annealing temperature were found for maximal amplification and specificity. This method was used for the amplification of conserved regions of pol and env genes. 1.5 pg of MT-2 and 5 pg of C3/44 Mo cell line DNAs were detected using nested-PCR and liquid hybridization in the pol system. The env system could detect 1.5 pg of MT-2 and 1.5 pg of C3/44 Mo cell lines DNAs using nested-PCR and liquid hybridization. The pol system can type both HTLV-I and HTLV-II in only two steps without the use of type-specific radiolabeled probes. Furthermore, this method can detect and discriminate the two viruses in one step PCR using the primers used in the nested-PCR. Nevertheless, there is a decrease in sensitivity of 100-fold. The results of five seropositive samples confirmed by Western blot are compared with PCR. PCR typed one of these samples as HTLV-I and the rest as HTLV-II. This technique is useful in cases such as window period, perinatal studies and when serologic results are not satisfactory.

[Isolation, sequencing and ultrastructure of HTLV-I and HTLV-II retroviruses. Presence of the HTLV-II subtype b among Spanish intravenous drug addicts]. / Aislamiento, secuenciación y ultraestructura de los retrovirus HTLV-I y HTLV-II. Presencia del HTLV-II subtipo b entre adictos a drogas por vía parenteral españoles.

BACKGROUND: Six cases of HTLV-I/II infection were selected for isolation and characterization of these retrovirus. METHODS: Detection of anti-HTLV antibodies was carried out by enzyme immunoassay (EIA), immunofluorescence (IFI), and Western blot (WB). Analysis of proviral DNA was performed by PCR. Viral culture and partial sequencing of the pol and pX genes were carried out. Electron microscopy morphologically characterized the viral particles. RESULTS: Serologic study demonstrated four cases of HTLV-II, one of HTLV-I, and one non-typeable HTLV infections. This last case was confirmed as positive for HTLV-II by PCR. Five new HTLV-II and one HTLV-I infected cell lines have been established by co-culture. Electron microscopy allowed morphologic characterization of the viral particles found in the infected cells. The sequence of the five strains of HTLV-II was identical demonstrating a divergence of 0.49% in the pX region and of 4.5% in the pol region compared with the HTLV-II Mo prototype. Comparison of these sequences with those corresponding to different strains of HTLV-II isolates from American Indians (subtypes b) suggest that these Spanish strains are more closely related with the subtype b than with the subtype a (HTLV-II Mo). Genetic variability study did not reveal any change in the sequence of these stains suggesting that the variability of these retroviruses in very infrequent in the regions studied. The analysis of the pol region of the HTLV-I strain demonstrated a divergence of 3.4% with respect to the sequence of the ATK-1 prototype (Japan) and of 1.7% of the strain HS-35 (Caribbean) showing a greater relation with the Caribbean strains than with those from Japan. CONCLUSIONS: The presence of HTLV-II subtype has been confirmed among intravenous drug addicts in Spain. Isolation and characterization of the HTLV-I strain demonstrated that this also circulating around Spain despite its South American origin.

Comparison of four methods for screening of cytomegalovirus antibodies in normal donors and immunocompromised patients.

Three commercial methods (enzyme immunoassay, fluoroimmunoassay and latex agglutination) and a complement fixation test were compared for detection of cytomegalovirus antibodies using a panel of 490 serum samples from blood and organ donors, from immunocompromised patients and paired sera from five patients with recent cytomegalovirus infection. An indirect immunofluorescence test for antibodies to cytomegalovirus was used for classifying samples giving discrepant results by any of the four methods. All methods showed high sensitivity and specificity, but the enzyme immunoassay and the latex agglutination tests had the highest sensitivity. Latex agglutination is recommended for large-scale screening of cytomegalovirus antibodies in blood and organ donors. Negative results obtained by latex agglutination should be confirmed by sensitive enzyme immunoassays.