OBJECTIVETo understand the distributions of DNA and neutralizing antibodies of human papillomavirus (HPV)16, 18 in 18-45 year-old women.

METHODSTotally, 1494 women were enrolled through multistage random sampling in Funing, Jiangsu province. Cervical exfoliated cells were collected from them for HPV DNA testing, and serum samples were taken from them for the detection of HPV16, 18 neutralizing antibodies by using pseudovirion-based neutralization assay(PBNA).

RESULTSAmong the 1494 women, 28(1.9%) and 188(12.6%) were positive for DNA and neutralizing antibody of HPV16 respectively, and 15(1.0%) and 60(4.0%) were positive for DNA and neutralizing antibody of HPV18, respectively. There were no significant differences in the detection rates of DNA and neutralizing antibody of HPV16, 18 among different age groups. About 16.7% of the women were infected with HPV16, 18, or both.

CONCLUSIONIn Funing county of Jiangsu province, most women aged 18-45 years has no immunity to HPV16 and 18, indicating that they are appropriate targets for HPV 16/18 vaccination.

Adolescent ; Adult ; Antibodies, Neutralizing ; isolation & purification ; Antibodies, Viral ; isolation & purification ; China ; DNA, Viral ; isolation & purification ; Female ; Human papillomavirus 16 ; immunology ; Human papillomavirus 18 ; immunology ; Humans ; Middle Aged ; Papillomavirus Infections ; prevention & control ; Papillomavirus Vaccines ; Young Adult

Seborrheic keratosis (SK) is a benign epidermal tumor of unknown etiology. Because of its wart-like morphology, human papillomavirus (HPV) has been suggested as a possible causative agent. Viral involvement, however, has not been confirmed yet despite extensive research. The aim of this study was to evaluate the presence of HPV 6/11, 31, 33 DNA in nongenital SK. We analyzed 40 biopsy specimens taken from patients with nongenital SK using in situ polymerase chain reaction (PCR) and PCR with tissue extracts. The SK specimens (n=40), analyzed by in situ PCR, were negative for all HPV probes tested (types 6/11, 31, 33). Control slides (condyloma acuminatum, n=3) were positive for type 6/11, 31, and 33 HPV probes tested. Melasma samples (n=4), the negative controls, were consistently negative. No HPV DNA band was detected by PCR with the tissue extracts from paraffin-embedded SK samples, while condyloma acuminatum, the positive controls, showed DNA bands of the correct molecular weights. Our results show that HPV type 6/11, 31, and 33 cannot be recognized as causative agents for nongenital SK, which is in contrast to the previous studies. Further studies are required to reveal the presence of other types (more than 90) of HPV DNA.
DNA, Viral/*analysis ; Female ; Human ; Keratosis, Seborrheic/*virology ; Male ; Papillomavirus, Human/*isolation & purification ; Polymerase Chain Reaction

Objective: To explore the effect of vaginal micro-environment alterations and HPV16 infection and their interaction in the progression of cervical intraepithelial neoplasia. Methods: The participants of this study came from the cervical lesions study cohort in Shanxi province, including 623 women with normal cervical (NC), 303 patients with pathogenically diagnosed low-grade cervical intraepithelial neoplasia (CINⅠ) and 93 patients with pathogenically diagnosed high-grade cervical intraepithelial neoplasia (CINⅡ/Ⅲ). The data of the demographic characteristics of the study subjects and factors related to cervical intraepithelial neoplasia were collected, and HPV16 infection were detected by using flow-through hybridization technology and H(2)O(2), β-glucuronidase, clotting enzyme, neuraminidase and leucocyte esterase in vaginal secretions were detected by using the combined detection kit of aerobic vaginitis and bacterial vaginosis. pH value and vaginal cleanliness were also detected at the same time. The database was established and analyzed by SPSS statistical software (version 22.0). Results: The HPV16 infection rate (trend χ(2)=55.45, P<0.001) and the abnormal rates of H(2)O(2) (trend χ(2)=26.19, P<0.001), pH (trend χ(2)=5.06, P=0.024), vaginal cleanliness (trend χ(2)=19.55, P<0.001), β-glucuronidase (trend χ(2)=17.52, P<0.001) and neuraminidase (trend χ(2)=14.90, P<0.001) increased gradually along with the severity of cervical intraepithelial neoplasia, but the abnormal rates of clotting enzyme and leucocyte esterase showed no same trend. The results of GMDR model analysis showed that there was interaction between HPV16 infection and abnormalities of H(2)O(2), β-glucuronidase, clotting enzyme and neuraminidase in CINⅠ group, and the interaction between HPV16 infection and the abnormalities of vaginal cleanliness, H(2)O(2), β-glucuronidase and neuraminidase in CIN Ⅱ/Ⅲ group. Conclusion: Our findings indicated that the vaginal micro-environment alterations and HPV16 infection could increase the risk of cervical intraepithelial neoplasia, and they might have an important synergistic effect in the progression of cervical intraepithelial neoplasia.
Female ; Human papillomavirus 16/isolation & purification* ; Humans ; Hydrogen Peroxide ; Papillomavirus Infections/virology* ; Uterine Cervical Neoplasms/virology* ; Uterine Cervical Dysplasia/virology*

OBJECTIVETo compare the effectiveness of immunofluorescence and polymerase chain reaction (PCR) in detecting human papilloma virus (HPV) in condyloma acuminata (CA).

METHODSHPVs in CA tissues from 60 patients were detected by immunofluorescence and PCR, respectively. Different subtypes of HPVs were also identified with restriction fragment length polymorphism (RFLP).

RESULTSThe positive detective rates of immunofluorescence and PCR were 56.67% (34/60) and 96.67% (58/ 60), respectively (P < 0.01). RFLP results showed HPV6 and HPV11 were the main subtypes in the detected virus, which accounted for 98.28%.

CONCLUSIONThe sensibility of PCR is superior to that of immunofluorescence.

Condylomata Acuminata ; virology ; Female ; Human papillomavirus 11 ; genetics ; isolation & purification ; Human papillomavirus 6 ; genetics ; isolation & purification ; Humans ; Male ; Microscopy, Fluorescence ; Polymerase Chain Reaction

Further understanding of male human papillomavirus (HPV) infection is necessary to prevent infection in men, as well as transmission to women. In our current study, we investigated patterns of HPV infection and genotype distributions in male genital warts using the Anyplex II HPV28 Detection kit. We reviewed the medical records of 80 male patients who presented to 5 neighborhood clinics in Ulsan, Korea, for the treatment of genital warts between April 2014 and January 2015. All patients underwent HPV genotyping. The prevalence and characteristics of HPV infection were analyzed, and the patterns of HPV infection according to age were assessed. Among the study patients, 13 (16.3%) were negative for HPV infection, 46 (57.3%) were infected with low-risk HPV, and 21 (26.3%) were infected with high-risk HPV. Patients with multiple HPV infection were more likely to have high-risk HPV infection (P = 0.001). The prevalence of HPV infection was much higher in samples obtained by tissue excision due to a definite lesion (P = 0.001). There were no differences in high-risk HPV infection (P = 0.459), multiple HPV infection (P = 0.185), and recurrence at diagnosis (P = 0.178) according to age. HPV-6 and HPV-11 were the most common type overall (39.7% and 13.8%, respectively). HPV-16 and HPV-18 were the most common high-risk infections (both 3.4%). HPV infection is not only commonly encountered in male genital warts, but is also accompanied by high-risk HPV and multiple infections.
Adult ; Condylomata Acuminata/epidemiology/*pathology/virology ; DNA, Viral/genetics/metabolism ; Genotype ; Human papillomavirus 11/*genetics/isolation & purification ; Human papillomavirus 16/genetics/isolation & purification ; Human papillomavirus 18/genetics/isolation & purification ; Human papillomavirus 6/*genetics/isolation & purification ; Humans ; Male ; Middle Aged ; Prevalence ; Real-Time Polymerase Chain Reaction ; Republic of Korea/epidemiology ; Retrospective Studies ; Risk Factors

OBJECTIVETo observe human papillomavirus (HPV) infection in women with cervical lesions in Huzhou area of Zhejiang province.

METHODS720 samples of cervical secretion or exfoliated cells were collected from women with cervical lesion in Huzhou area. Human papillomavirus was detected by suspension array technique.

RESULTSPositive HPV infection was detected in 25.42% cases (183/720), with 135 cases of single HPV type, 33 of dual HPV types and 15 of multiple HPV types. HPV16 and HPV58 were the most prevalent types in 183 HPV positive cases.

CONCLUSIONThe most prevalent high-risk types of HPV are HPV16 and HPV58 in Huzhou area of Zhejiang province.

Adolescent ; Adult ; Alphapapillomavirus ; classification ; isolation & purification ; China ; Female ; Human papillomavirus 16 ; isolation & purification ; Humans ; Middle Aged ; Papillomavirus Infections ; virology ; Uterine Cervicitis ; virology ; Young Adult

OBJECTIVETo determine the different rates of human papillomavirus types 6 (HPV-6) and 11 (HPV-11) infection in biopsy samples from pointed condyloma patients, and to construct prokaryotic expression system of the major capsid protein L1 of the virus so as to establish an ELISA for detecting the expression of L1 gene in the biopsy samples.

METHODSUsing a double PCR based on the L1 gene of HPV-6 and HPV-11, the infection rates of HPV-6 and HPV-11 in the biopsy samples were determined. The whole length of HPV-6 L1 gene was amplified using PCR and the target amplification fragment was sequenced after T-A cloning. The prokaryotic expression system pET32a-L1-E. coli BL21 (DE3) was constructed and SDS-PAGE was used to measure the expression of the target recombinant protein rL1. Rabbit anti-rL1 serum was prepared and immuno-diffusion assay was applied to examine the antiserum titer. ELISA was established to detect the expression of L1 gene in the biopsy samples.

RESULTSIn the biopsy samples from 116 pointed condyloma patients, 92.2% (107/116) were detectable for HPV-6 and/or HPV-11. Of the 107 positive samples, 70.1% (75/107) and 23.4% (25/107) were positive for HPV-6 or HPV-11 alone and 6.5% (7/107) were coinfected with both HPV-6 and HPV-11 respectively. When compared with the reported corresponding sequences, the homology of nucleotide and sequence of the cloned HPV-6 L1 gene was from 99.20% - 99.93% while its putative amino acid sequence homology was from 99.80% - 100%, suggesting IPTG could induce the expression of rL1. The immuno-diffusion titer of the rabbit anti-rL1 serum was 1:4. 88.8% (103/116) of the biopsy samples were the major capsid protein L1 detectable.

CONCLUSIONA prokaryotic expression system of HPV-6 L1 gene, a double PCR assay for HPV-6 and HPV-11 genotyping, and an ELISA assay for detecting the major capsid protein L1 were successfully established in this study. The pointed condyloma patients in Zhejiang area mainly infected with HPV-6. The HPV in the focus frequently expressed major capsid protein L1.

Animals ; Biopsy ; Capsid Proteins ; genetics ; Condylomata Acuminata ; pathology ; virology ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Human papillomavirus 11 ; genetics ; isolation & purification ; Human papillomavirus 6 ; genetics ; isolation & purification ; Humans ; Papillomavirus Infections ; virology ; Polymerase Chain Reaction ; Rabbits ; Sequence Homology, Nucleic Acid

A simple, rapid and sensitive colorimetric loop mediated isothermal amplification (LAMP) method was established to detect HPV6 and HPV 16 respectively. The method employed a set of four specially designed primers that recognized six distinct sequences of HPV6-E6 or HPV16-E7 for amplification of nucleic acid under isothermal conditions at 63 degrees C for one hour. The amplification process of LAMP was monitored by the addition of HNB (hydroxy naphthol blue) dye prior to amplification. A positive reaction was indicated by a color change from violet to sky blue and confirmed by real-time turbidimeter and agarose electrophoresis. Thirteen cervical swab samples having single infection with 13 different HPV genotypes were examined to evaluate the specificity. A serial dilution of a cloned plasmid containing HPV-E6 or HPV-E7 gene was examined to evaluate the sensitivity. The results showed that no cross-reaction with other HPV genotypes was observed. The colorimetric LAMP assay could achieve a sensitivity of 1000 copies, 10-20 times lower than that of real-time PCR. The assay was further evaluated with 62 clinical specimens and consistent results were obtained compared with the detection using Kai Pu HPV Genotyping Kit. We concluded that this colorimetric LAMP assay had potential usefulness for the rapid screening of the HPV6 or HPV16 infection in the laboratories and hospitals of provincial and municipal region in China.
Colorimetry ; methods ; DNA Primers ; chemistry ; genetics ; Genotype ; Human papillomavirus 16 ; genetics ; isolation & purification ; Human papillomavirus 6 ; genetics ; isolation & purification ; Humans ; Nucleic Acid Amplification Techniques ; instrumentation ; methods ; Papillomavirus Infections ; virology

OBJECTIVETo evaluate the value of high-risk HPV (hrHPV) detection by Hybrid Capture II (HC2) in screening cervical intraepithelial neoplasm (CIN).

METHODSTotally 723 patients who had received a dual screening with thinprep cytologic test (TCT) and HC2 in our department were analyzed retrospectively. Among them, 350 patients received a triple examination with TCT, HC2, and colposcopic biopsy.

RESULTSAmong the 723 patients, the incidences of hrHPV infection with atypical squamous cell (ASC), low squamous intraepithelial lesion, and high squamous intraepithelial lesion were 70.7% (94/133), 88.9% (249/280), and 90.9% (90/99), respectively, significantly higher than 55.5% (117/211), the incidence of hrHPV infection with normal cytological results (P = 0.005, P < 0.001, P < 0.001, respectively). Among 350 cases who were received triple examination, the incidence of hrHPV infection with cervical intraepithelial neoplasia (CIN) 1 and CIN 2 were 88.9% (72/81) and 96.3% (52/54), significantly higher than 77.7% (153/197), the incidence of hrHPV infection with normal pathological results (P = 0.03, P = 0.002); The incidence of hrHPV infection with CIN 3 and squamous cancer were 91.7% (11/12) and 100.0% (6/6), also higher than normal cases. Among these 350 cases, the incidence of hrHPV infection with ASC was 79.3% (69/87). The incidence of CIN 2-3 with ASC and hrHPV infection was 38.0%, significantly higher than the incidence of CIN 2-3 with ASC and without hrHPV infection (5.9%) (P = 0.04).

CONCLUSIONhrHPV infection has a close relation with CIN, and the incidence of hrHPV infection increases along with the severity of CIN.

Adolescent ; Adult ; Cervical Intraepithelial Neoplasia ; virology ; Cervix Uteri ; pathology ; virology ; Female ; Human papillomavirus 16 ; isolation & purification ; Human papillomavirus 18 ; isolation & purification ; Humans ; Middle Aged ; Nucleic Acid Hybridization ; methods ; Papillomavirus Infections ; epidemiology ; virology ; Uterine Cervical Neoplasms ; virology

Adult ; Carcinoma ; virology ; Cervical Intraepithelial Neoplasia ; virology ; Female ; Human papillomavirus 16 ; isolation & purification ; Human papillomavirus 18 ; isolation & purification ; Humans ; In Situ Hybridization ; Middle Aged ; Papillomavirus Infections ; Uterine Cervical Neoplasms ; virology