Oral antibodies to human papillomavirus type 16 in women with cervical neoplasia

This study investigated the relationship between human papillomavirus type 16 (HPV-16) antibodies detected in oral fluid from women with cervical neoplasia, their HPV-16 antibody seroprevalence, and their cervical HPV-16 DNA presence. Cervical HPV-16 DNA was detected by polymerase chain reaction in 43.2% (35/81) of these women. The prevalence of IgG and IgA antibodies to HPV-16 virus-like particles (VLP-16) in oral fluid and was investigated by enzyme-linked immunosorbent assay. Anti-VLP-16 IgA antibodies were detected in oral fluid from 54.3% (44/81) of women with cervical neoplasia, compared with 8% (3/36) in controls (P = 0.000002). Anti-VLP-16 IgG was detected in oral fluid from 43.2.9% (25/72) and 13.3% (4/30; P = 0.029), respectively. Women who were HPV-16 DNA positive at their cervical lesion, displayed an oral fluid anti-VLP-16 IgA prevalence of 60.7% (17/28) and HPV-16 DNA negative women an oral fluid anti-VLP-16 IgA prevalence of 50% (20/40; P = 0.38). Oral fluid anti-VLP-16 IgG prevalence in HPV-16 DNA positive women was 28.6% (8/28) compared with 40% (16/40) in oral fluid from HPV-16 DNA negative women (P = 0.3). Amongst HPV-16 DNA positive women, the anti-VLP-16 IgG seroprevalence was 75% (21/28) and IgA seroprevalence 35.7% (10/28) and for the HPV-16 DNA negative women these values were 60% (24/40) and 32.5% (13/40), respectively. Oral IgA antibody testing proved no more sensitive than serum antibody detection for the determination of HPV infection but could be useful as a non-invasive screening method for women with cervical neoplasia and for estimating the mucosal antibody response to HPV vaccines.     

Cervical mucus antibodies against human papillomavirus type 16, 18, and 33 capsids in relation to presence of viral DNA.

To investigate whether cervical mucus antibodies against human papillomavirus (HPV) capsids are associated with the detection of HPV DNA or HPV-related cytological diagnoses, 611 samples of cervical secretions from 359 women referred to a colposcopy clinic were tested by an enzyme-linked immunosorbent assay for the presence of immunoglobulin A (IgA) antibodies against HPV capsids of HPV type 16, 18, or 33 and for the presence of cervical HPV DNA by PCR. Among subjects with at least one cervical sample positive for HPV type 16 (HPV-16) DNA, 28.1% also had at least one HPV-16 IgA-positive cervical sample (odds ratio [OR] = 2.9; P = 0.0003). IgA to HPV-18 was also more common among HPV-18 DNA-positive subjects (OR = 3.1; P = 0.0325) and IgA to HPV-33 was more common among HPV-33 DNA-positive subjects (OR = 4.2; P = 0.0023). Cervical IgA antibodies to HPV-16 were also more common among patients with cervical intraepithelial neoplasia, particularly among patients with cervical intraepithelial neoplasia grade I (P < 0.0005). The data indicate that an HPV type-restricted IgA antibody response against HPV capsids is detectable in cervical mucus and is associated with a concomitant cervical HPV infection.     

Frequency of antibodies against E4 and E7 from human papillomavirus type 16 in Mexican soldiers

Summary. The high prevalence of HPV in men’s genitalia and the low frequency of virus-associated lesions gave rise to questions on the influence of infection-site on the HPV antibody profile. In a cross-sectional study, HPV infection in penis and urethra, and serum antibodies against HPV-16 E4 and E7 proteins were evaluated in 288 Mexican soldiers. The results showed that HPV prevalence was 31% (51% in penis, 11% in urethra and 38% in both sites), while 47% were multiple infections. Overall, seroprevalence was 13% for anti-E4 antibodies and 6% for anti-E7. However, the highest prevalence of anti-E4 antibodies was observed in men with HPV infection in urethra (30%), while for E7 antibodies, the highest prevalence (10%) was found in men who tested positive for HPV in penis. The prevalence of IgG and IgA anti-E4 was related to HPV-16 urethral infection, while detection of HPV-16 in penis was related to IgG anti-E7 prevalence. In conclusion, the high-risk sexual behavior observed in this population might be responsible for high HPV prevalence and multiple infections. However, the seroprevalence of E4 and E7 was similar to that observed in healthy Mexican women. These results suggest that the humoral immune response against HPV infection in men differs, depending on the site of infection.     

Cervical human papillomavirus (HPV) infection in South African women: implications for HPV screening and vaccine strategies

The prevalence of cervical human papillomavirus (HPV) in South African women (n = 1,073) increased from 20.4% (173/848) in women with normal cytology to 41.7% (48/115) in women with atypical squamous cells of undetermined significance, 70.2% (40/57) in women with low-grade squamous intraepithelial lesions, and 83% (44/53) in women with high-grade squamous intraepithelial lesions (HSILs). HPV types 16 and 35 were the dominant types in women with HSILs but not in women in the other categories.     

Prevalence of antibodies to human papillomavirus (HPV) type 16 virus-like particles in relation to cervical HPV infection among college women.

A human papillomavirus type 16 (HPV-16) virus-like particle (VLP)-based enzyme-linked immunosorbent assay (ELISA) was used to measure serum antibody to capsid proteins in 376 sexually active college women who were also screened for the presence of genital HPVs by PCR and interviewed for demographic and behavioral risk factors for HPV infection. The seroprevalence was 46% in women with HPV-16 DNA in the genital tract. The corresponding values for women who harbored other HPV types or no HPV in the genital tract were 30 and 19%, respectively (HPV-16 group versus no-HPV group; odds ratio [OR], 3.7; 95% confidence interval [CI], 1.5 to 8.9). The antibody response was significantly higher among women with a high viral load than among those with a low viral load (median optical density value, 0.838 versus 0.137, P = 0.009). Comparable levels of seroreactivity were observed among women infected with HPV types distantly or closely related genetically to HPV-16. Seroreactivity was significantly associated with an age of 25 to 30 years (OR, 2.3; 95% CI, 1.2 to 4.4), three or more lifetime sexual partners (OR, 2.9; 95% CI, 1.1 to 10), and history of a sexually transmitted disease other than HPV (OR, 3.1; 95% CI, 1.5 to 6.3). The percent seropositivity increased linearly with number of lifetime sexual partners until reaching a plateau at 35% for women with more than six partners (chi for linear trend, P < 0.001). The low sensitivity of HPV-16 VLP-based ELISA may limit the usefulness of the assay as a diagnostic test for HPV-16 infection. However, the assay appears to have adequate specificity and should be useful as an epidemiological marker of HPV-16 infection and sexual behavior.     

HIV‐1 seroconversion promotes rapid changes in cervical human papillomavirus (HPV) prevalence and HPV‐16 antibodies in female sex workers

The extent to which human immunodeficiency virus (HIV‐1) infection impacts on the ability to mount an effective immune response to HPV is unknown, but is relevant in planning HPV vaccine strategies for HIV‐1 infected individuals. This longitudinal study investigated changes shortly after HIV‐1 seroconversion on cervical HPV types and HPV‐16 antibody responses in serum and at the cervix of female sex workers. Typing of HPV DNA from cervical cells was done prior to HIV‐1 seroconversion and within 1 year and greater than 2 years after HIV‐1 seroconversion. Antibody determinations on serum and cervico‐vaginal rinse samples were by HPV‐16 virus‐like particle‐based, enzyme‐linked immunosorbent assay. Of 104 women tested, 40 (38.4%) became HIV‐1 seropositive (HIV‐positive) during the course of the study. Shortly after HIV‐1 seroconversion a significant increase in multiple (>1) HPV infection (OR 4.0, 95% CI 1.3–11.9) was observed compared with HIV‐1 seronegative (HIV‐negative) women and certain changes in HPV type infection. HIV‐1 seroconversion resulted in a reduced prevalence of serum HPV‐16 IgA and cervico‐vaginal IgA and IgG but an increased prevalence of serum HPV‐16 IgG. All HIV‐positive women had been exposed to HPV‐16 as all displayed serum HPV‐16 IgG. Serum HPV‐16 responses were maintained at a high magnitude in the presence of HPV‐16 infection irrespective of HIV infection, but decreased in the absence of HPV‐16 infection. In conclusion, HIV‐1 seroconversion in sex workers rapidly increased cervical HPV infection and caused a reduced ability to produce cervical HPV‐16 antibodies but a continued ability to generate serum IgG antibodies. J. Med. Virol. 81:203–210, 2009. © 2008 Wiley‐Liss, Inc.     

Prevalence of Chlamydia pneumoniae and Mycoplasma pneumoniae Immunoglobulin G and A Antibodies in a Healthy Finnish Population as Analyzed by Quantitative Enzyme Immunoassays

Chlamydia pneumoniae and Mycoplasma pneumoniae immunoglobulin G (IgG) and IgA antibody seroprevalence rates and antibody levels related to age and gender were studied. The samples (n = 742) were collected during a nonepidemic period and analyzed by quantitative enzyme immunoassays (EIAs). Seroprevalence to C. pneumoniae was found to increase sharply in young children, and in the 15- to 19-year-old group it reached levels as high as 70 and 60% for IgG and IgA, respectively. After adolescence, seroprevalence showed a transient decrease and then continued to increase, although less dramatically than in early childhood. In the elderly the seroprevalence of IgG antibodies reached 75 and 100% in women and men, respectively. The corresponding rates of IgA antibodies were 73 and 100%. When a randomly selected subgroup of samples (n = 66) was analyzed in parallel by a microimmunofluorescence test and an EIA for C. pneumoniae IgA antibodies, similar seroprevalence rates were obtained (36 versus 35%). Seroprevalence to M. pneumoniae was already found to increase very sharply in 2- to 4-year-old children, reaching 16% for IgG and 8% for IgA. Seroprevalence to M. pneumoniae also continued to increase in adolescence, but in contrast to that to C. pneumoniae, the increase leveled off at about 40 to 50% in adulthood. In subjects aged over 65 years, prevalence did not exceed 60% for IgG or 35% for IgA. The seroprevalence patterns as well as the medians and variations of levels of C. pneumoniae and M. pneumoniae IgG antibodies were similar to those of corresponding IgA antibodies. Compared to IgG antibodies, IgA antibodies do not seem to be of additional value in the diagnosis of infections caused by these pathogens when single serum specimens are studied.     

Cytological physiognomies and genotype distribution of human papillomaviruses among HPV/HIV co-infected and HPV mono-infected women

BackgroundCo-infection of High Risk Human Papillomavirus (HR-HPV) and HIV is thought to favour initiation of intraepithelial squamous cell lesion and subsequent progression to cervical carcinoma.ObjectivesEvaluation of cytological physiognomies in relation to possible age influence and the genotype distribution of human papillomaviruses among HPV/HIV co-infected and HPV monoinfected women in Kisii, Kenya.MethodsThe case-control study enrolled 42 HPV/HIV co-infected and 42 HPV monoinfected women. Cervical swabs were collected in ThinPrep vials for HPV tying and cytological analysis. HPV subtypes were assayed by Xpert® HPV system (GXHPV-CE-10).ResultsMono-infected women aged 30–39 years had the highest proportion of low grade squamous intraepithelial lesion (LSIL) at 14 (16.67%) while the co-infected aged 50–59 years had the highest proportion of high grade squamous intraepithelial lesion (HSIL) at 9 (10.71%). HPV-16 genotype was the most predominant and it increased with age rise. Older coinfected and mono-infected women (>40 years) had HSIL and LSIL as the most predominant cytological grade respectively.ConclusionThe predominance of HPV-16 and HPV-18/45 genotypes in the study setting is a consideration that would benefit targeted prophylactic vaccination programs. HPV testing and cervical cancer screening for young and older women on a regular basis ought to be reinforced.     

Human papillomavirus genotype spectrum in Czech women: correlation of HPV DNA presence with antibodies against HPV-16, 18, and 33 virus-like particles.

Because the biological spectrum of human papillomavirus (HPV) genotypes present in cervical cancer lesions varies according to the geographical region studied, and because little genotype information is available for Central and Eastern European countries, we studied the endemic HPV-genotype spectrum in cervical samples collected from women visiting gynaecological departments of selected hospitals in the Czech Republic. In a series of 389 samples, 171 (44.0%) were positive for HPV DNA using a consensus-primer polymerase chain reaction (PCR). Genotyping of the HPV PCR products was done using dot-blot hybridisation with type-specific oligonucleotide probes and thermocycle DNA sequencing. Twenty-two different HPV types were detected, HPV-16 being the most prevalent type irrespective of severity of the lesions (55.0%). Multiple HPV types were found in 16.4% of our HPV-DNA-positive samples. The prevalence of HPV infection was 23.0% in women with normal findings and 59.4% in patients with cervical neoplasia, and increased significantly with the severity of the disease: 52.9% in low-grade lesions, 58.0% in high-grade lesions, and 73.5% in cervical carcinomas (P for trend < .00001). In the sera of 191 subjects, 89 with normal findings and 102 with different forms of cervical neoplasia, the prevalence of HPV-specific IgG antibodies was tested by an enzyme-linked immunosorbent assay (ELISA) using virus-like particles (VLPs) of HPV-16, -18, and -33. Antibodies were significantly more prevalent in HPV-DNA-positive than in HPV-DNA-negative women and there was no association with age. In agreement with the results of HPV genotyping, antibodies reactive with HPV-16 VLPs were the most frequent and, moreover, their prevalence increased with the cervical lesion severity. About half of the subjects with smears in which either HPV-16 or HPV-33 DNA had been detected possessed antibodies reactive with homotypic VLPs. With HPV-18-DNA-positive subjects, however, fewer than 25% displayed homotypic antibodies. In general, subjects older than 30 years of age had antibodies reactive to HPV-specific VLPs more often than subjects younger than 30 years of age. In women with benign findings, the seropositivity to HPV-16, -18, and -33 VLPs increased with age, whereas in women with cervical neoplasia the seropositivity decreased with age.     

Polymer-based enzyme-linked immunosorbent assay using human papillomavirus type 16 (HPV16) virus-like particles detects HPV16 clade-specific serologic responses

Human papillomavirus type 16 (HPV16) virus-like particles (VLP) were used as antigen in a polymer enzyme-linked immunosorbent assay (ELISA) to measure antibodies to HPV capsid proteins. Serum samples from 575 college women, previously tested for the presence of cervicovaginal HPV DNA, were analyzed. The prevalences of anti-HPV16 VLP antibodies at baseline were 14.1% for immunoglobulin G (IgG) and 6.4% for IgA. The seroprevalences of IgG in women with cervicovaginal HPV16, HPV16-related types, and other HPV types were 55, 33, and 19%, respectively (P < 0.001), compared to the prevalence in women without an HPV infection (10%). HPV VLP IgA seropositivity was associated with high HPV16 VLP IgG optical density values. The seropositivity of IgG antibodies was independently associated with infection with HPV16 or HPV16-related types, increased number of lifetime male partners for vaginal sex, having sex with men >/= 5 years older, history of abnormal PAP smear, older age, and living separately from parents. Use of HPV16 VLP polymer ELISA detects clade-specific responses and suggests an HPV16 VLP vaccine may have broader protection that initially anticipated.     

HPV genotype prevalence in cytologically abnormal cervical samples from women living in south Italy

Human papillomavirus (HPV) infection is the commonest sexually transmitted infection, and high-risk HPV types are associated with cervical carcinogenesis. This study investigated: the HPV type-specific prevalence in 970 women with an abnormal cytological diagnosis; and the association of HPV infection and cervical disease in a subset of 626 women with a histological diagnosis. HPV-DNA was researched by nested PCR/sequencing and the INNOLiPA HPV Genotyping assay. The data were analysed by the chi-square test (p相似文献   

Association of HPV infection and Chlamydia trachomatis seropositivity in cases of cervical neoplasia in Midwest Brazil

High-risk human papillomavirus (HPV) is considered the main etiological agent for cervical neoplasia. However, the presence of a single type HPV infection alone is unlikely to be sufficient to cause cervical cancer. There is epidemiologic evidence suggesting that HPV and Chlamydia trachomatis play a central role in the etiology of cervical intraepithelial neoplasia and subsequent cervical cancer. To evaluate the HPV prevalence and the seropositivity for C. trachomatis in women referred to the colposcopy clinic due to an abnormal cervical smear and to examine the effect of this association on the severity of cervical neoplasia. Following enrollment, 131 patients underwent colposcopy and biopsies when necessary. HPV DNA was detected by the polymerase chain reaction (PCR) and genotyping was performed by reverse line-blot hybridization assay. C. trachomatis seropositivity was tested by ELISA for the detection of IgG antibodies. The prevalence of HPV infection was 86.3%. Seropositivity for C. trachomatis was 26%. Thirty-one women (27.4%) were positive for C. trachomatis antibodies and HPV-DNA. The most prevalent HPV type in C. trachomatis-seropositive women were HPV 16 (51.6%) and this HPV type was present mainly in neoplasia cases. Positivity for HPV, particularly HPV types 16 and 18, and C. trachomatis seropositivity was significantly associated with a diagnosis of high grade neoplasia. Borderline significance was observed after adjustment for HPV. C. trachomatis seropositivity is associated with high grade neoplasia in women infected with HPV, mainly when the types 16 and 18 were involved.     

Age distribution of antibodies to human papillomavirus in children,women with cervical intraepithelial neoplasia and blood donors from South Africa

Sera from 95 women with cervical intraepithelial neoplasia (CIN), 95 age-matched female blood donors, and 155 children aged between 1 and 12 years were tested by enzyme-linked immunosorbent assay (ELISA) for levels of serum IgG to three human papillomavirus (HPV) peptides (HPV-16 E2 [E2-16], HPV-18 E2 (E2-18], HPV-16 L1 [L1-16]), as well as HPV-16 virus-like particles (VLP-16) and bovine papillomavirus type 1 virus-like particles (BPV-VLP). In the adult group antibodies to E2-16 and VLP-16 were significantly associated with CIN when compared to the blood donor controls (P = .039 and P = .002, respectively). In women with CIN there was an increase in seropositivity to E2-16 and a decrease in seropositivity to VLP-16 with age. Antibodies to HPV-16 E2 could therefore be an important marker of CIN in women over 40 years of age, whereas antibodies to VLP-16 could be a marker for CIN in younger women. There was no correlation with CIN and antibodies to E2-18, L1-16, and BPV-VLP. In the children's sera antibodies were detected to E2-16 (44.5%), E2-18 (18.7%), L1-16 (20%), VLP-16 (4.5%), · and BPV-VLP (5.1%). Between the ages of 3 and 12 years the prevalence of antibodies to E2-16 decreased with age. The presence of antibodies to HPV-16 in young children indicated infection with either HPV-16 or a related virus. HPV DNA isolation from children could help resolve this question. J Med Virol 51:126–131, 1997. © 1997 Wiley-Liss, Inc.     

厦门地区女性人乳头瘤病毒感染状况调查

目的 对厦门地区妇女宫颈人类乳头瘤病毒（human papilloma virus,HPV）亚型进行筛查,以探讨其分布规律.方法 采用凯普医用核酸分子快速杂交仪,对7683名2013年1月-2013年12月到厦门市174医院妇科门诊或病房就诊的女性进行生殖道21种HPV感染基因亚型筛查.结果 7683例样本中,HPV感染者1421例,阳性率18.50％.感染人群主要集中在30～39岁.其中高危型HPV阳性率为16.01％,低危型HPV阳性率为2.49％.单一基因型别者1148例,阳性率为14.94％;双重感染者220例阳性率为2.86％;三重以上感染者53例,阳性率为0.70％;HPV感染阳性率居前6位的基因型分别为HPV-52（3.64％）、HPV-16 （3.33％）、HPV-58（2.98％）、HPV-53（2.16％）、HPV-CP8304（1.91％）和HPV-18（1.09％）.结论 厦门地区妇女HPV感染率高,且以高危型单一基因亚型感染为主;感染基因型别主要以HPV-52、16、58、53、CP8304和18为主,具有一定的地域差异性;52及58型感染率高,对于疫苗的研制和开发有参考意义.     

Pre- and posttreatment serum antibody responses to HPV 16 E2 and HSV 2 ICP8 proteins in women with cervical carcinoma.

Serum antibodies to early proteins of human papillomavirus type 16 (HPV 16 E2 protein) and herpes simplex virus type 2 (HSV 2 ICP8) can be measured by ELISA. In the serum of 122 newly diagnosed cervical carcinoma patients and age-matched controls, enhanced IgA antibody levels to an HPV-16 E2 protein derived peptide no. 245 indicated a 9.5-fold (95% confidence limits 2.8-57.2) relative risk of cervical carcinoma. No significant risk was found with a corresponding HPV 6 E2 peptide or HSV 2 ICP8. To evaluate the HPV 16 E2 peptide as a possible tumor marker for cervical carcinoma serial postoperative serum samples were tested from 27 women with cervical carcinoma. Antibody responses to the HPV 16 E2 peptide depended on the clinical stage. Stage I and II patients showed decreasing posttreatment IgA and/or IgG antipeptide antibody levels. Stage III and IV patients initially showed decreasing antipeptide antibody levels followed by increasing levels. These patients also showed increasing IgG antibody levels to the HSV 2 ICP8. However, increasing antibody levels to the HPV 16 E2 peptide indicated significantly (P less than 0.05) worse 2-year disease free survival (recurring disease) than did stable or decreasing antibody levels. The results suggest that serum antipeptide antibodies to the HPV 16 E2 peptide no. 245 can be used for the monitoring of cervical carcinoma.     

Abundance of Multiple High-Risk Human Papillomavirus (HPV) Infections Found in Cervical Cells Analyzed by Use of an Ultrasensitive HPV Genotyping Assay

PCR methods enable the detection of a large variety of human papillomavirus (HPV) genotypes that infect the anogenital tract. However, PCR with consensus primers, general primers, and, to a lesser extent, broad-spectrum primers may underrepresent the true prevalence of HPV, especially the true prevalence of multiple infections. We compared the rate of HPV positivity determined by a broad-spectrum PCR with primers BSGP5+ and BSGP6+ (BS-PCR) coupled to an established bead-based multiplex HPV genotyping (MPG) assay with the rate of HPV positivity determined by a multiplex PCR with type-specific primers (TS-PCR) coupled to a newly developed MPG assay for 735 selected cervical scraping samples. While the primers used for the BS-PCR are located within the L1 region of the HPV genome, the primers used for the TS-PCR target the E7 gene. The overall rates of positivity for the 19 HPV types included in both assays were 60.9% and 72.2% by the BS-PCR and the TS-PCR, respectively, and the two assays found multiple infections in 34.8% and 58.0% of the specimens, respectively. Both HPV detection assays allowed the semiquantitative detection of HPV types and identified the same dominant HPV type in 66.6% of the multiple infections. In conclusion, the TS-PCR-MPG assay significantly increased the rate of detection of HPV DNA and the number of infections with multiple HPV types detected and demonstrated that the prevalence of low-copy-number HPV infections in the anogenital tract may be strongly underestimated by conventional HPV amplification methods, especially in cases of multiple infections. As a consequence, PCR-TS-MPG appears to be highly suited for analysis of the significance of multiple infections in the development of cervical cancer and for the study the natural history and the latency of HPV.Human papillomaviruses (HPV) are DNA viruses that infect cutaneous and mucosal epithelia. Until now, approximately 100 HPV genotypes have been fully characterized on the basis of the isolation of complete genomes (7), and there is evidence that a larger number exist (1). There are approximately 45 known mucosal HPV types; and these are further divided into three groups on the basis of their epidemiological association with cervical cancer: high-risk HPV (Hr-HPV) types (types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 73, and 82), putative high-risk HPV (pHr-HPV) types (types 26, 53, and 66), and low-risk HPV (Lr-HPV) types (e.g., types 6, 11, 40, 42, 43, 44, and 70) (18). Hr-HPV types are causally associated with several malignant diseases, of which cervical cancer has particular significance, being the second most common cancer in women worldwide and the main cancer of women in most developing countries (18). Hr-HPV type DNA has been detected in 99.7% of cervical cancer tissue specimens (26), and persistent infection with an oncogenic HPV type, particularly HPV type 16 (HPV-16) or HPV-18, is recognized as a necessary cause of cervical cancer. HPV genotyping is of importance for the study of the natural history of infections with one or several HPV types and the role of HPV persistence in the progression of cervical lesions and for the monitoring of vaccine efficacy.Among HPV-positive women, 20 to 40% harbor in their cervices at least two types that were acquired simultaneously or successively (17). It remains controversial whether an infection with multiple types (referred to here as a multiple infection) is a risk factor for the persistence of HPV and for cervical lesions (20, 21). Moreover, it remains unknown whether women with, e.g., quadruple infections are at higher risk than women with double infections. Interest in multiple HPV infections has recently increased as prophylactic vaccines against HPV types 6, 11, 16, and 18 are expected to also provide partial protection against related HPV types by cross-neutralizing antibodies (12). Therefore, it is important to accurately type all HPV infections present in one patient. It will also be of particular interest to study the long-term impact of vaccination on the established equilibrium in the distribution of HPV types within immunized populations. Therefore, the sensitive, reliable, and unbiased profiling of the individual HPV types in patients with multiple infections is important to learn more about the natural history of HPV and to evaluate the effect of HPV vaccination.HPV typing based on PCR methods has continuously been improved over the past few years. One of the most common PCR uses the GP5+ and GP6+ primer pair, which targets conserved sequences within the L1 region of the virus genome flanking highly variable type-specific sequences (6). Use of PCR with this primer pair allows the amplification of a broad range of mucosal HPV types in a single reaction. The HPV genotype can be determined by analyzing the PCR product generated by sequence analysis, restriction fragment length polymorphism analysis, or hybridization with type-specific probes in different formats, such as the reverse line blot (RLB) assay (24) or the bead-based multiplex HPV genotyping (MPG) method (22). Recently, the performance of the PCR with primers GP5+ and GP6+ has been improved by addition of eight forward broad-spectrum primers and two backward broad-spectrum primers (primers BSGP5+ and BSGP6+ [BS-PCR]), which has led to a more homogeneous analytical sensitivity for the detection of genital HPV types (23). Consequently, the detection of HPV types 30, 39, 42, 44, 51, 52, 53, 68, 73, and 82, as well as the detection of multiple infections, was significantly improved by BSGP5+/BSGP6+ MPG. In addition, it has been shown that not only the use of degenerate primers (primers GP5+ and GP6+) (6) and/or consensus primers (primers MY09 and MY11) (2) but also the use of broad-spectrum primers (14) may lead to the underdetection of low copy numbers of HPV, particularly in cases of multiple infections (19).Recently, a highly sensitive, multiplex type-specific HPV E7 PCR system detecting a larger proportion of multiple infections than GP5+/GP6+ RLB has been described (9). However, the detection of PCR products was based on an array primer extension (APEX) assay, a time- and labor-intensive procedure with little high-throughput ability.In this report, we describe the development of an ultrasensitive and highly specific Luminex-based assay for the genotyping of products from the multiplex type-specific E7 PCR (TS-PCR). The results obtained by this novel high-throughput assay were compared to those obtained by the already established, sensitive BSGP5+/BSGP6+ MPG assay.     

Epidemiology and risk factors for human papillomavirus infection in a diverse sample of low-income young women

BackgroundTwo HPV vaccines prevent infection with HPV-16 and HPV-18, high-risk (cancer-associated) HPV types which together cause approximately 70% of cervical cancers; one vaccine also prevents HPV-6 and HPV-11, which together cause approximately 90% of anogenital warts. Defining type-specific HPV epidemiology in sexually experienced women will help estimate the potential clinical benefits of vaccinating this population.ObjectivesTo examine HPV epidemiology in a diverse sample of sexually experienced women, and to determine factors associated with high-risk HPV and vaccine-type HPV (HPV-6, HPV-11, HPV-16 and HPV-18).Study designCross-sectional study of 13–26-year-old women (N = 409) who completed a questionnaire and provided a cervicovaginal swab. Swabs were genotyped for HPV using PCR amplification. Logistic regression models were used to determine whether participant characteristics, knowledge, and behaviors were associated with high-risk and vaccine-type HPV.ResultsMost women (68.4%) were positive for ≥1 HPV type, 59.5% were positive for ≥1 high-risk type, 33.1% were positive for ≥1 vaccine-type HPV, and 3.5% were positive for both HPV-16 and HPV-18: none was positive for all four vaccine types. In adjusted logistic regression models, Black race (OR 2.03, 95% CI 1.21–3.41) and lifetime number of male sexual partners (OR 4.79, 95% CI 2.04–11.23 for ≥10 partner vs. ≤1 partner) were independently associated with high-risk HPV infection.ConclusionsHPV prevalence was very high in this sample of sexually active young women, but <5% were positive for both HPV-16 and HPV-18, suggesting that vaccination could be beneficial for many individual women who are sexually experienced.     

Sourcing of the WHO human papillomavirus type 18 international standards for HPV antibody levels

BackgroundHPV serology is important for studies of vaccine immunogenicity, but can not be performed in a comparable manner without international standardisation.ObjectivesTo find suitable candidate sera from naturally infected persons for use as International Standards (IS) for antibodies to high-risk HPVs, with priority for HPV-18.Study design946 healthy Thai women (median age 44, range 18–83) and 61 cervical cancer patients were screened using an HPV pseudovirion-Luminex assay to detect antibodies to genital (HPV-6,-11,-16,-18,-31,-33,-45,-52,-58,-68) and non-genital HPV types (HPV-5,-15,-32,-38 and -76). Suitable candidate sera should ideally be mono-specific (have reactivity against only one genital HPV) and have high antibody levels that are stable over time.ResultsSeroprevalences of HPV-16,-31,-52 and -58 were at least twice as high among cancer patients compared to healthy individuals. Thirteen healthy women who met the IS inclusion criteria in initial testing also consented to blood-bag donations. Donations from 2 women with high HPV-18 Ab titers were pooled to the HPV-18 candidate IS, later established as the WHO official IS for HPV antibodies. Sera that could potentially be used as candidate IS for other oncogenic HPVs have also been identified.ConclusionsIn the Thai population, seroepidemiology implicated HPV types HPV-16,-31,-52 and -58 as particularly associated with cervical cancer. A well characterized cohort study has allowed sourcing of materials for an IS for HPV-18 antibodies and could conceivably be used for IS for other HPV types as well.     

Influence of HIV-1 and/or HIV-2 infection and CD4 count on cervical HPV DNA detection in women from Senegal,West Africa

BackgroundHIV infection is associated with greater risk of precancerous lesions and cervical cancer in women. However, several factors remain unclarified regarding the association between HIV infection and HPV detection, especially among those with HIV type 2 versus type 1 infection and severely immunocompromised persons.ObjectivesTo evaluate HPV overall and type-specific detection among HIV-infected and uninfected women in Senegal.Study designDetection of HPV DNA for 38 genotypes in cervical swabs using PCR-based methods was evaluated in HIV-positive (n = 467) and HIV-negative (n = 2139) women participating in studies in Senegal. Among HIV-1 and/or HIV-2 positive women, CD4 counts were assessed. Adjusted multivariable prevalence ratios (PR) were calculated.ResultsThe prevalence of any HPV DNA and multiple HPV types was greater among HIV-infected individuals (78.2% and 62.3%, respectively) compared with HIV-negative women (27.1% and 11.6%). This trend was also seen for HPV types 16 and 18 (13.1% and 10.9%) compared to HIV-negative women (2.2% and 1.7%). HIV-infected women with CD4 cell counts less than 200 cells/μl had a higher likelihood of any HPV detection (PR_a 1.30; 95% CI 1.07–1.59), multiple HPV types (PR_a 1.52; 95% CI 1.14–2.01), and HPV-16 (PR_a 9.00; 95% CI 1.66–48.67), but not HPV-18 (PR_a 1.20, 95% CI 0.45–3.24) compared to those with CD4 counts 500 cells/μl or above.ConclusionHIV-infected women, especially those most severely immunocompromised, are more likely to harbor HPV. Measures to prevent initial HPV infection and subsequent development of cervical cancer through focused screening efforts should be implemented in these high risk populations.     

Predominance of serum antibodies to synthetic peptide stemming from HPV 18 open reading frame E2 in cervical adenocarcinoma.

AIMS: To determine if there are type specific differences in serum antibody responses to synthetic peptides derived from human papillomavirus (HPV) open reading frame (ORF) E2 in patients with cervical carcinoma. METHODS: Diagnostic phase sera from 88 age-matched women with cervical adenocarcinoma (AC), cervical squamous cell carcinoma (SC), ovarian cancer (OC) or no gynaecological malignancy were available. Serum IgG and IgA antibodies to synthetic peptides corresponding to a residue of HPV 6, 11, 16, and 18 ORF E2 18 amino acids long and a control peptide from mumps virus were determined by ELISA. RESULTS: Both IgA and IgG antibody positivity to the HPV 18 peptide were associated with increased risk (9.0-fold, confidence limits 1.5-199) for AC. IgA positivity to HPV 11, 16, and 18 peptides was associated with an increased risk for SC. However, the association of IgG antibodies to HPV 16 peptide with SC was not significant. IgA or IgG antibodies to HPV 6 or mumps virus peptides were not associated with increased risk for AC, SC, or OC. CONCLUSIONS: These results suggest a specific role for HPV 18 in AC. Differences in antibody responses to HPV peptide in AC and SC suggest immunopathogenetic differences between the two types of cervical carcinoma.