Primer-mediated Polymerase Chain Reaction Research Articles

Viral Load in the Natural History of Human Papillomavirus Infection Among Men in Rural China: A Population-based Prospective Study.

High-risk human papillomavirus (HR-HPV) load is predictive of HR-HPV persistence and subsequent carcinogenesis in women. However, in men, data on genital HPV load and its effect on the natural history of HPV infection are limited. The subjects included 1532 men aged 25-65 years with up to 7 biannual visits for evaluation of genital HPV load in rural China during 2009-2013 who were positive for ≥1 of the 18 selected HPV types (including 10 HR-HPV types) detected by general primer-mediated polymerase chain reaction (PCR) and sequencing. Type-specific HPV load was quantified with real-time PCR and dichotomized based on median values. Men with multiple lifetime sex partners were more likely to have higher overall levels of HR-HPV load across visits (adjusted odds ratio, 2.42; 95% confidence interval [CI], 1.12-5.24; 2 partners vs 0-1 partner). Higher levels of HR-HPV load at the time of the first HPV diagnosis conferred an increased probability of the subject remaining type-specific HPV-positive up to 12 months and an increased probability of persistent/intermittent infection (virus detected repeatedly with or without a period of intercurrent negativity) versus transient infections (single-time positive). Higher overall HR-HPV levels were predictive of reduced HR-HPV clearance rates (adjusted hazard ratio, 0.47; 95% CI, .27-.83). Having multiple lifetime sex partners is associated with increased male genital HR-HPV load. Higher HR-HPV load predicts persistence of HR-HPV in men from rural China.

Detection of human papilloma virus in esophageal cancer in Xinjiang Kazakh patients by general primer-mediated polymerase chain reaction

Detection of human papilloma virus in esophageal cancer in Xinjiang Kazakh patients by general primer-mediated polymerase chain reaction

A noncommercial polymerase chain reaction–based method to approach one hundred percent recombinant clone selection efficiency

Molecular cloning is an important procedure in molecular biology, but this is often a rate-limiting step and can be very time-consuming, possibly due to low ligation efficiency. Here, we describe a simple polymerase chain reaction (PCR)-based strategy to approach 100% selection efficiency. The post-ligation mixture containing the recombinant was subjected to insert-specific primer-mediated PCR amplification using a high-fidelity DNA Pfu polymerase generating a plasmid containing staggered nicks. The PCR mixture was then digested with endonuclease DpnI, which digests the methylated and hemimethylated parental DNA template. The nicked vector was transformed into XL1 blue supercompetent cells where the nicks were repaired, thus amplifying and selecting only the newly amplified recombinant clones.

Clinical and molecular prognostic factors in operable laryngeal cancer

Many factors affect the prognosis in operable laryngeal squamous cell carcinoma (LSCC). Many clinical factors have been implicated in tumor recurrence and poor survival of the patients. The aim of the present study is to investigate the demographic, clinical and histological characteristics as prognostic factors. Moreover, our aim is to analyze the role of modern molecular biomarkers in the prognosis of patients with LSCC. One hundred patients with operable laryngeal carcinoma underwent surgery as primary treatment between April 1999 and April 2002. Ninety-four of them were men and 6 women, with a median age of 62 years (39-77). All demographic data of the patients were recorded. Staging of the tumor revealed 20 cases with T2 cancer, 46 cases with T3 and 34 cases with T4, while N classification included 91 patients with N0 tumor, 3 with N1 and 6 with N2. Among the 100 cases, 47 were located in the glottis, 46 in the supraglottic region and 7 were transglottic. Histology grading revealed 35 cases of grade G1, 50 cases of G2 and 15 cases of G3. Postoperatively, all patients were followed regularly for the possibility of tumor relapse, with a median follow-up period of 40.2 months (4.8-58.4). During the operation, a tissue specimen was collected from the tumor. The specimens were used for RNA and DNA extraction. Isolated RNA was used to investigate the expression of wt-p53, bcl-2, VEGF and EGFR by the reverse transcriptase PCR method (RT-PCR) using specific primers, while genomic DNA was used for the detection of EBV and HPV (16/18 subtypes) by the consensus primer-mediated polymerase chain reaction method (PCR). All data such as tumor recurrence and survival were recorded. Statistical analysis was performed using the SPSS and STATA statistical packages in order to investigate the role of all clinical and molecular factors and their combinations as significant prognostic markers. The tumor recurrence rate was 31%, while the tumor associated death rate was 27% and total death rate 30%. Univariate analysis for overall survival showed significance for the T stage, TNM stage and site of the tumor. Univariate analysis for the time to progression showed significance for the T stage, N stage, TNM stage, site of the tumor and tumors simultaneously positive for EGFR and VEGF, while EGFR expression was borderline insignificant. Multivariate analysis revealed TNM stage as the only significant factor for overall survival, and TNM stage, site of the tumor and EGFR expression as significant factors for time to progression. The molecular biomarkers EGFR and VEGF have a prognostic significance in laryngeal cancer in addition to the established clinical prognostic factors such as the stage and site of the tumor. These markers, apart from their role in carcinogenesis, seem to play an important role in tumor relapse.

Does human papillomavirus play a role in the development of bladder transitional cell carcinoma? A comparison of PCR and immunohistochemical analysis

Aim: To investigate the role of human papillomavirus (HPV) in the development of bladder transitional cell carcinoma (TCC). Methods: Seventy eight paraffin wax embedded TCC samples were tested for the...

Human Papillomavirus Infection Among Women with Normal Cervical Pap Smear Tests

ABSTRACTVarious human papillomaviruses (HPVs) have been strongly implicated as etiological agents in human cervical cancer. The primary method for detection of high-risk HPVs is still the Papanicolaou-stained (Pap) smear. However, the Pap smear has some limitations, including high false-negative rates. Using consensus and specific primer-mediated polymerase chain reaction (PCR), we investigated 55 women with normal cervical cytology. HPV DNA was found in 11 women (20%), including HPV-16 in 6 cases (10.9%), HPV-6 in 3 cases (5.5%), HPV-6/18 in 1 case (1.8%) and HPV-6/33 in 1 case (1.8%). The obtained results demonstrate the importance of careful follow-up and repeated testing for HPV persistence. Moreover, these findings raise the possibility that HPV diagnostics by molecular virology techniques might be useful as an adjunct to Pap smear screening.

Human papillomavirus type 16 and 18 DNA in oral squamous cell carcinoma and normal mucosa

The aim of this study was to ascertain the prevalence of HPV 16/18 DNA in oral squamous cell carcinoma (OSCC) vs. normal oral mucosa, and to correlate the virologic data with other factors obtained from the patients' records. One hundred and thirteen paraffin embedded tissue samples (73 OSCC and 40 normal oral mucosa) were studied using HPV type specific primer-mediated polymerase chain reaction (PCR). Seventy-four per cent (54/73) of OSCC and 55% (22/44) of normal oral mucosa were positive for HPV 16/18 DNA. Statistical analysis indicated there was a significant difference between HPV16/18 positive OSCC vs. normal oral mucosa (P=0.040), and that age (<60 years) and gender (male) were correlated with the presence of HPV16/18 in the tumour. No significant association was found between the presence of HPV and other risk factors, including tobacco use, alcohol use, tumour location, histologic grade or TNM staging. We found a significant association of HPV16/18 with oral squamous cell carcinoma. Also, HPV16/18 is a co-factor in oral carcinogenesis, particularly in male patients and patients under the sixth decade. In addition, we found that HPV infection is a common event in the normal oral mucosa.

Co-factors related to the causal relationship between human papillomavirus and invasive cervical cancer in Honduras.

A case-control study was conducted in Honduras to identify co-factors in the carcinogenic pathway by which human papillomavirus (HPV) causes invasive cervical cancer. Ninety-nine cases aged 23-65 (median 47) years participated. Two controls were matched to each case by age and clinic where they first presented for cytological screening; controls had no cervical abnormalities. Information on risk factors was obtained by personal interviews in the clinics regarding sociodemographic, reproductive and behavioral characteristics. Human papillomavirus was detected in cervical scrapes by general primer-mediated polymerase chain reaction (PCR) followed by sequence analysis to identify the different types present. All cases had squamous cell tumours and most were FIGO (International Federation of Gynecologists and Obstetricians) class II or higher; HPV was strongly associated with cervical cancer (odds ratio [OR] = 7.66, 95% CI : 3.88-15.1). Among HPV-positive women, dose-response relationships were observed for education, age at first intercourse and exposure to wood smoke that persisted after adjustment for previous screening. Among HPV-negative women, the number of sexual partners and parity were associated with cervical cancer. The protective effect of previous cytological screening operated independently of HPV. Our findings speak for the powerful role that both primary and secondary education plays in fostering a lifestyle that reduces the risk of invasive cervical cancer. The data suggest that important elements of such a lifestyle include later age at first sexual intercourse, a limited number of pregnancies, greater likelihood of undergoing cytological screening and reduced exposure to carcinogens in the household environment.

Human papillomavirus and prognoses of patients with cancers of the upper aerodigestive tract.

Some studies have shown that human papillomavirus (HPV) infection may play not only an etiologic role in anogenital cancers but also a role in the clinical outcome. The objective of the current study was to determine whether detection of HPV DNA in primary squamous cell carcinomas of the upper aerodigestive tract (UADT) is a prognostic factor in patients with the disease. The authors analyzed archival specimens of UADT tumors from 101 randomly selected patients with evaluable samples for HPV DNA detection. HPV testing was performed using a general primer-mediated polymerase chain reaction. The overall detection rate was 16.8% (17 of 101 specimens). HPV DNA was detected at higher rates in specimens from younger patients and in well-differentiated tumors. Pharyngeal tumors were more likely to be HPV positive (30.0%) than buccal (10.3%) or laryngeal tumors (15.4%), but the differences were not significant. The detection rate was similar for T1-T2 tumors (17.4%) and T3-T4 tumors (15.6%). However, tumors without lymph node metastasis were more likely to be HPV positive (21.4%) than tumors with lymph node involvement (6.5%). Kaplan-Meier and Cox regression survival analyses did not show any difference in overall or disease free survival according to HPV detection. Although the HPV DNA detection rate was slightly higher in local than in regionally spread tumors, our results support the hypothesis that it is very unlikely that HPV detection plays any role in the prognoses of patients with UADT squamous cell carcinoma.

Human papillomavirus and prognoses of patients with cancers of the upper aerodigestive tract

BACKGROUND Some studies have shown that human papillomavirus (HPV) infection may play not only an etiologic role in anogenital cancers but also a role in the clinical outcome. The objective of the current study was to determine whether detection of HPV DNA in primary squamous cell carcinomas of the upper aerodigestive tract (UADT) is a prognostic factor in patients with the disease. METHODS The authors analyzed archival specimens of UADT tumors from 101 randomly selected patients with evaluable samples for HPV DNA detection. HPV testing was performed using a general primer-mediated polymerase chain reaction. RESULTS The overall detection rate was 16.8% (17 of 101 specimens). HPV DNA was detected at higher rates in specimens from younger patients and in well-differentiated tumors. Pharyngeal tumors were more likely to be HPV positive (30.0%) than buccal (10.3%) or laryngeal tumors (15.4%), but the differences were not significant. The detection rate was similar for T1–T2 tumors (17.4%) and T3–T4 tumors (15.6%). However, tumors without lymph node metastasis were more likely to be HPV positive (21.4%) than tumors with lymph node involvement (6.5%). Kaplan–Meier and Cox regression survival analyses did not show any difference in overall or disease free survival according to HPV detection. CONCLUSIONS Although the HPV DNA detection rate was slightly higher in local than in regionally spread tumors, our results support the hypothesis that it is very unlikely that HPV detection plays any role in the prognoses of patients with UADT squamous cell carcinoma. Cancer 1999;85:1903–9. © 1999 American Cancer Society.

Different risk factor patterns for high-grade and low-grade intraepithelial lesions on the cervix among HPV-positive and HPV-negative young women.

Risk factors for cervical intraepithelial neoplasia have most often been studied in high-grade lesions. Furthermore, in a high proportion of the studies, human papillomavirus (HPV), the most significant risk determinant of cervical neoplasia, was not taken into account when evaluating other risk factors. To compare risk factors for ASCUS (atypical cells of undetermined significance), LSIL (low-grade squamous intraepithelial lesion) and HSIL (high-grade squamous intraepithelial lesion), we conducted a case-control study among 20 to 29 year-old women participating in a prospective cohort study in Copenhagen. It included 131 women with ASCUS, 120 women with LSIL, 79 women with HSIL and 1,000 randomly chosen, cytologically normal, control women. All participants had a personal interview and a gynecological examination including a Pap smear and cervical swabs for HPV DNA detection using general primer-mediated polymerase chain reaction. The most significant risk determinant of all 3 disease categories was the presence of genital HPV DNA. The risk factor pattern was nearly identical for ASCUS and LSIL, but differed significantly from that for HSIL. Stratified analysis by HPV-status showed that, apart from, respectively, smoking and parity among HPV-positive women, and smoking and number of sex partners among HPV-negative women, no additional risk factors were observed for ASCUS and LSIL. In contrast, among HPV-negative women with HSIL, long-term use of oral contraceptives was the most important risk factor. However, our result should be taken with great caution as it is based on very small numbers, and as it is unknown whether the HPV-negative lesions constitute a true entity. Among HPV-positive women, the risk of HSIL was associated with e.g., years of sex life without barrier contraceptive use, early age at first genital warts and smoking. Whether the risk factors that are applicable only to HSIL represent factors related to progression remains unknown.

Different risk factor patterns for high‐grade and low‐grade intraepithelial lesions on the cervix among HPV‐positive and HPV‐negative young women

Risk factors for cervical intraepithelial neoplasia have most often been studied in high-grade lesions. Furthermore, in a high proportion of the studies, human papillomavirus (HPV), the most significant risk determinant of cervical neoplasia, was not taken into account when evaluating other risk factors. To compare risk factors for ASCUS (atypical cells of undetermined significance), LSIL (low-grade squamous intraepithelial lesion) and HSIL (high-grade squamous intraepithelial lesion), we conducted a case-control study among 20 to 29 year-old women participating in a prospective cohort study in Copenhagen. It included 131 women with ASCUS, 120 women with LSIL, 79 women with HSIL and 1,000 randomly chosen, cytologically normal, control women. All participants had a personal interview and a gynecological examination including a Pap smear and cervical swabs for HPV DNA detection using general primer-mediated polymerase chain reaction. The most significant risk determinant of all 3 disease categories was the presence of genital HPV DNA. The risk factor pattern was nearly identical for ASCUS and LSIL, but differed significantly from that for HSIL. Stratified analysis by HPV-status showed that, apart from, respectively, smoking and parity among HPV-positive women, and smoking and number of sex partners among HPV-negative women, no additional risk factors were observed for ASCUS and LSIL. In contrast, among HPV-negative women with HSIL, long-term use of oral contraceptives was the most important risk factor. However, our result should be taken with great caution as it is based on very small numbers, and as it is unknown whether the HPV-negative lesions constitute a true entity. Among HPV-positive women, the risk of HSIL was associated with e.g., years of sex life without barrier contraceptive use, early age at first genital warts and smoking. Whether the risk factors that are applicable only to HSIL represent factors related to progression remains unknown. Int. J. Cancer 76:613–619, 1998.© 1998 Wiley-Liss, Inc.

Type I diabetic pregnancy and subclinical human papillomavirus infection.

It has been suggested that diabetic pregnancy is an immunosuppressive state. To determine whether the possible immunosuppression in pregnant diabetics might result in an increased risk for human papillomavirus (HPV) infection, we studied exfoliated cells from the uterine cervix, vagina, and posterior commissure of the vulva by means of dot blot hybridization with use of a probe cocktail of HPV types 11, 16, and 18 under low stringency and by means of consensus primer-mediated polymerase chain reaction (PCR) targeted to the HPV L1 and E1 regions. For this study, samples from 31 pregnant diabetics whose glucose levels had been reasonably well controlled were analyzed during the first trimester; samples from 27 of these patients were analyzed again during the third trimester. Fifty-one healthy pregnant women were included as controls. Only one of the pregnant diabetics was positive for HPV DNA. The L1 PCR products of the first and third trimester samples from this patient were sequenced, and both were found to represent HPV 61. Three of the pregnant controls were positive for HPV: two were positive for HPV 16, and one was positive for HPV 6. The 95% confidence limits for the prevalence of HPV were calculated to be 0.1%-16.7% for the diabetics and 1.2%-16.2% for the controls. The 95% confidence limits for the difference between the groups were -11.6%-6.3%. These results suggest that pregnant diabetics do not have an increased risk of developing HPV infection, at least when their glucose levels remain well controlled.

Human papillomavirus infection in progressive and non-progressive cervical intraepithelial neoplasia.

Human papillomavirus (HPV) infection is common in cervical intraepithelial neoplasia (CIN) and is widely held to be responsible for its progression to grade 3. This thesis is examined here. Comparison of the level of HPV changes in 133 lesions that had not progressed to that in those from 197 women with histologically proven CIN 3 failed to reveal significant differences in their level of HPV infection on cytology, histology or in situ hybridization. However, in both these groups, some of the cases that did not show HPV positivity on in situ hybridization with probes reacting with the common HPV types did show evidence of HPV DNA using a general primer-mediated polymerase chain reaction. This may indicate low-copy number infections or non-productive infections. Such reactions were more frequent in the women with progressive lesions, and it is probable that they may also have been at greater risk of cervical infection in general. The present findings suggest that a further factor, a cocarcinogen, may be involved in progression to CIN 3, HPV being a common forerunner, providing a proliferative environment and thus favoring such an event.

Prevalence of mucosotropic human papillomaviruses in squamous-cell carcinoma of the head and neck.

The prevalence of mucosotropic human papillomavirus (HPV) DNA in 63 squamous-cell carcinomas (SCC) from different anatomic sites in the head and neck was determined by general primer-mediated polymerase chain reaction (GP-PCR). HPV DNA was detected in 20.6% of SCC. Additional type-specific PCR for HPV 6, 11, 16, 18, 31 and 33 demonstrated the presence of HPV 16 alone in these carcinomas. HPV 16 was also detected in normal epithelium from the resection margins of the majority of HPV-positive SCC. HPV status did not correlate with tumour site, whether primary or recurrent, TNM stage, metastases, degree of differentiation, smoking or alcohol history, fate or survival.

5447839 Detection of human papillomavirus by the polymerase chain reaction : Manos M Michele; Bauer Heidi M; Greer Catherine E; Resnick Robert M; Ting Yi Richmond, CA, United States assigned to Hoffmann-La Roche Inc

Objective:To study the relationship between human papillomavirus (HPV) and cervix erosion,and to find the best condition of polymerase chain reaction (PCR).Methods:HPV in the cervix cells was detected by general primermediated PCR (GPPCR).Results:Fifty healthy women and 34 cervicitis were included in the study.The positive rates of HPV in healthy and cervicitis were 22.0% and 47.2% respectively (P005).Conclusion:Detecting HPV by GP-PCR is a sensitive,spesific,rapid and economic method and it was a method of early and fast diagnosis of virus infection and epidemiology study.

Human papillomavirus in vulvar and vaginal carcinoma cell lines.

A number of reports associate human papillomavirus (HPV) with cervical cancer and cancer cell lines derived from this tumour type. Considerably fewer reports have focused on the role of HPV in carcinomas from other sites of female anogenital squamous epithelia. In this study we have tested for the presence of HPV in eight low-passage vulvar carcinoma cell lines and one extensively passaged cell line, A431. One cell line from a primary vaginal carcinoma was included. The presence of the HPV was evaluated by the polymerase chain reaction (PCR), by Southern blot analysis and by two-dimensional gel electrophoresis. General primer-mediated PCR was applied by using primers from the L1 region, E1 region and HPV 16 E7 region. Southern blot hybridisation was performed under low-stringency conditions (Tm = -35 degrees C) using a whole genomic HPV 6/16/18 probe mixture and under high stringency conditions (Tm = -18 degrees C) with the whole genomic probes of HPV 16 and 33. HPV 16 E6-E7 mRNA was assessed by ribonuclease protection assay (RPA). HPV was found in only one vulvar carcinoma cell line, UM-SCV-6. The identified type, HPV 16, was integrated in the cell genome and could be amplified with all primers used. Also E6-E7 transcripts were found in these cells. Five original tumour biopsies were available from the HPV-negative cell lines for in situ hybridisation. All these were HPV negative with both the HPV 6/16/18 screening probe mixture under low stringency and the HPV 16 probe under high stringency. The results indicate that vulvar carcinoma cell lines contain HPV less frequently than cervical carcinoma cell lines and suggest that a significant proportion of vulvar carcinomas may evolve by an HPV-independent mechanism.

Correlation of human papillomavirus 16 and 18 with prognostic factors in invasive cervical neoplasias

Forty-seven patients with cervical carcinoma were examined in order to correlate human papillomavirus (HPV) types with prognostic factors in invasive cervical neoplasias. Age, clinical stage, histological type, and grade and parity were analysed with respect to HPV status as determined by a general primer mediated polymerase chain reaction (PCR) or a type specific PCR. All but one sample (98%) harboured HPV sequences: HPV 16 was found in 26 cases (55%), HPV 18 in 19 cases (40%), and HPV 31 in 1 case. The presence of HPV 18 DNA was significantly associated with cancers developed below 40 years of age (P = 0.029). HPV 18 detection was associated with poor differentiation malignancy (P = 0.045) and histological types of poor prognosis (adenocarcinoma or nondifferentiated carcinoma; P = 0.006). HPV 18 positivity was also correlated with advanced clinical stages (FIGO II and III; P = 0.032). Parity and HPV status proved to be independent of each other (P approximately 0.99). Eighty-seven percent (27/31) of pelvic lymph nodes from HPV positive patients contained HPV DNA. The virus types found in lymph nodes were identical with those of the primary tumours in all cases. Virological results were compared to those obtained by routine histological examination. Only 6 of 27 patients with HPV positive lymph nodes had any histological evidence of metastasis. Nevertheless, the lack of metastasis as detected by histology does not exclude the possibility of relapses. Follow-up of the clinical prognostic significance of PCR detection of HPV in the possible sites of early metastases.

General primer-mediated polymerase chain reaction for simultaneous detection and typing of human papillomavirus DNA in laryngeal squamous cell carcinomas

The aim of this study was to investigate the prevalence of different human papillomavirus (HPV) types in laryngeal squamous cell carcinomas using general primer-mediated polymerase chain reaction (PCR). Tumour sections from 42 patients with laryngeal carcinomas were investigated. For HPV DNA amplification, consensus primers were used which were directed to the L1 coding region of the HPV genome. Analysis of the PCR products was done using 2% agarose gel electrophoresis followed by restriction enzyme analysis to identify different HPV types. Amplification of the human TGF-beta 1 DNA was successfully performed in 36/42 (85.7%) of samples confirming the presence of sufficient DNA for viral amplification. HPV DNA was detected in 8/36 (22.2%) of the tumours examined (three HPV-6, two HPV-16, one HPV-11, two unknown HPV types). HPV DNA was not detected in any of the non-neoplastic laryngeal mucosa which was used as control (n = 15). Fifty per cent of women had HPV-positive tumours compared with 8% of men (chi2 = 5.8, P < 0.05). Our data indicate that while the overall prevalence of HPV in laryngeal carcinomas is fairly high (22.2%), the frequency of high-risk types (HPV-16 & HPV-18) is low (5.5%). HPV probably acts as a promoter in the multistep process of carcinogenesis in squamous mucosal cells of the larynx.

Human papillomavirus DNA and anti-HPV secretory IgA antibodies in cytologically normal cervical specimens.

Cervical specimens collected from 163 cytologically healthy women were screened for the presence of human papillomavirus (HPV) DNA and anti-HPV secretory IgA antibodies. HPV DNA was detected by a general primer mediated polymerase chain reaction (PCR), which amplifies a conserved region from the L1 ORF of genital HPVs. The PCR products were typed by restriction enzyme digestion. A total of 35 samples (21.5%) were positive for HPV DNA (13 samples for HPV 6, 6 for HPV 16, 3 for HPV 18, and 13 for untypeable HPV X). HPV DNA positivity was significantly higher among women under 25 years of age (34.8%) than among the older patients (12.4%) (P < 0.001). An enzyme-linked immunosorbent assay (ELISA) using synthetic peptide antigens was carried out to detect local secretory IgA antibodies against the following HPV specific antigens: HPV 16 E2, HPV 16 E7, HPV 16 L1, HPV 16 L2, and HPV 11 L2. Thirty-four secretions (20.9%) were found to react with at least one of the oligopeptides. Anti-HPV IgA positivity was the highest among women aged 25-32 years, and it was significantly lower in both the younger and the older age groups (P < 0.05). Correlation between HPV DNA and anti-HPV IgA detection was rather weak: anti-peptide IgA positivity was 34.3% (12 of 35) among HPV DNA positive patients compared to 17.2% (22 of 128) among HPV DNA negative women (P < 0.05). The fluctuating course of latent HPV infections should be considered in evaluating the low level of correlation between HPV DNA and anti-HPV IgA positivity.