OBJECTIVETo evaluate the diagnostic applicability of human papillomavirus (HPV) liquid chip assay which is based on Luminex XMAP System, and perform a HPV epidemiologic study with the liquid chip in women of Shandong province.

METHODSTo detect HPV genotypes on a 96-well plate with the liquid chip which can simultaneously detect and identify 26 common HPV genotypes in a total of 2925 cervical scrapes obtained from gynecological outpatients as well as to analyze the relationship between HPV types and different cervical diseases by studying the distribution of HPV genotypes and pathologic diagnosis.

RESULTSAmong 639 cases who performed pathologic/cytological and histological diagnoses, 184 cases are in group of normal cytology, 266 cases in group of, 77 cases in group of cervical intra-epithelial neoplasia (CIN) I, 7 cases in group of CIN I - II, 46 cases in group of CIN I - II, 46 cases in group of CIN I - II and 13 cases in group of cervical cancer. The overall incidence of HPV in our samples is 36.0% (1054/2925) and 23 types of all 26 types on liquid chip are found. The most common genotypes found are HPV-16 (26.75%), HPV-52 (25.75%), HPV-58 (10.47%), HPV-18 (8.87%) and HPV-11 (6.94%). Among all the positive types, 87.32% are high-risk HPV and 13.68% are low-risk HPV genotypes. Both single and multiple types are easily identified, showing 66.22% ( n = 698) single type and 33.78% ( n = 356) multiple types. Of all the 1054 HPV-positive cases, 261 (24.8%) is occupied by women 21 to 25 years of age and progressively lower by older age groups, reaching 4.9% by women between 51 to 67 years old. The incidence of HPV in our samples is 23.37%, 33.08%, 54.54%, 57.14%, 82.61%, 91.30% and 100% for normal cytology, inflammation,CIN I ,CIN I - II, CIN II ,CIN III, and carcinomas specimens, respectively. Infections with more that one virus are common, accounted for 4.89%, 7.14%, 18.18%, 28.57%, 41.30%, 43.37% and 38.46% for normal cytology, inflammation, CIN I, CIN I - II, CIN II, CIN III, and carcinomas specimens, respectively. Based on the criteria of histology and pathology, the sensitivity, specificity, positive-predictive value and negative-predictive value of HPV liquid chip assay for detecting all cases of CIN II, III are 88.57%, 76.63%, 68.89% and 92.16% respectively. Conclusion The common types of HPV infection are 16, 52, 58, 18, 11, 6, 56 and 31. The HPV-positive rate increased along with the increase of grading on cervical lesions. There are more younger women among all the HPV-positive ones. Multiplex HPV genotyping by liquid chip appears to be highly suitable for diagnostic screening as well as the conduction of large-scale epidemiological studies.

Adolescent ; Adult ; Aged ; Cervical Intraepithelial Neoplasia ; epidemiology ; virology ; China ; epidemiology ; Female ; Gammapapillomavirus ; classification ; genetics ; Genotype ; Human papillomavirus 11 ; classification ; genetics ; Human papillomavirus 16 ; classification ; genetics ; Human papillomavirus 18 ; classification ; genetics ; Human papillomavirus 6 ; classification ; genetics ; Humans ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; methods ; Papillomaviridae ; classification ; genetics ; Papillomavirus Infections ; epidemiology ; virology ; Uterine Cervical Neoplasms ; epidemiology ; virology ; Young Adult

BACKGROUND: The human papillomavirus (HPV) is the most common etiologic factor of cervical cancer. It was reported that the incidence of cervical intraepithelial neoplasia and cervical carcinoma was increased when normal women was infected with HPV. To date, for detection and classification of HPV, it were used by hybridization method using the DNA probe specific for HPV and in situ hybridization method for fixed paraffinized tissue, but reported that these methods were inadequate for detection of HPV DNA because of low sensitivity and complex procedure. Compared with these methods, polymerase chain reaction (PCR) was reported as a highly sensitive molecular biologic technique which could detect the HPV DNA in the cervical epithelial cell of women. Thus we used PCR for the investigation of detection rate of HPV 16 and 18, and its relationship with Pap smear class in normal women. METHODS: In 86 normal women, we had extracted the HPV DNA from cervical swab samples and then detected the presence of HPV DNA by nested PCR. RESULTS: In the cases of 86 normal women, the detection rate for HPV DNA was about 7.0%. In the cases of 26 women with Pap smear class I, the HPV DNA was not detected. And in the cases of 60 women with Pap smear class II, the detection rate for HPV DNA was about 10.0%; HPV subtype 16 about 6.7%; HPV subtype 18 about 1.7%; and the coinfection rate of HPV subtype 16 and 18 about 1.7%. CONCLUSIONS: We think that women who was previously infected with high-risk HPV should be examined for Pap smear test in regular time interval, and if the result of Pap smear was abnormal, then should be examined for the presence of the HPV DNA.
Cervical Intraepithelial Neoplasia ; Classification ; Coinfection ; DNA ; Epithelial Cells ; Female ; Human papillomavirus 16 ; Humans* ; In Situ Hybridization ; Incidence ; Papillomavirus Infections* ; Paraffin ; Polymerase Chain Reaction* ; Uterine Cervical Neoplasms

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

OBJECT: To support the application of the Bethesda classification to the histologic diagnosis of cervical squamous lesions. METHODS: We evaluated the cervical biopsies and human papilloma virus (HPV) DNA status of 100 clinic patients. The 100 biopsy specimens were analyzed using criteria of the Bethesda system for low-grade squamous intraepithelial lesion(LSIL) and high-grade squamous intraepithelial lesions(HSIL) in association with analysis of HPV DNA virus type by PCR. RESULTS: By PCR amplified from paraffin embedded tissues, the 40 of 90 cases of classified as HSIL were HPV 16 or 18 positive and 1/10 cases of classified as LSIL was HPV 16 positive. CONCLUSION: This study supports the usefullness of certain histologic criteria of the Bethesda system for distinguishing squamous intraepithelial lesions into two grades.
Biopsy ; Cervix Uteri ; Classification ; Diagnosis* ; DNA ; DNA Viruses ; Female ; Human papillomavirus 16* ; Humans ; Papilloma ; Paraffin ; Polymerase Chain Reaction

To investigate E6 and E7 gene variations of human papillomavirus type 16 in Yunnan Province, DNA was extracted from 2000 gynecological outpatient samples. For Human papillomavirus (HPV) genotyping, the genomic DNA was first amplified by the consensus MY09/MY11 primer pair followed by nested PCR with GP5+/GP6+ primers, then the PCR products were subjected to direct DNA sequencing. A total of 20 HPV-16 viral DNAs were identified. E6 and E7 genes of HPV-16 viral DNA were then amplified using E6 and E7 specific primers, the PCR products were purified and sequenced. The results showed that mutations were found at nucleotide position 178 of HPV-16 E6 gene in 10 cases,the mutation rate was 50%; For HPV-16 E7 gene, the mutations were found at nucleotide position 647 in 10 cases; the mutation rate was 50%. Phylogenetic analysis showed that Asian (As) variants of HPV-16 were predominated in Yunnan, China. None of African-1, African-2 variants of HPV-16 was found in this region.
Adult ; Base Sequence ; China ; Female ; Genetic Variation ; Human papillomavirus 16 ; classification ; genetics ; isolation & purification ; Humans ; Middle Aged ; Molecular Sequence Data ; Mutation ; Oncogene Proteins, Viral ; genetics ; Papillomavirus E7 Proteins ; genetics ; Papillomavirus Infections ; virology ; Phylogeny ; Repressor Proteins ; genetics

AIMThere is an increasing evidence for the role of high risk human papillomavirus (HPV) in the pathogenesis of oral squamous cell carcinoma (OSCC). The purpose of this study is to evaluate the relevance of HPV infection to the survival and prognosis of OSCC.

METHODOLOGYFifty-two patients with OSCC were followed from 4 to 88 months with a median of 50.7 months. HPV DNA was identified in formalin-fixed, paraffin-embedded tumor specimens by nested PCR with MY09/MY11 and GP5+/GP6+ primer pairs and the HPV genotype was determined by direct DNA sequencing. Association between the HPV status and risk factors for cancer as well as tumor-host characteristics were analyzed. Survival curves were calculated by the Kaplan-Meier method and analyzed using the log-rank test.

RESULTSHPV was found in 40.4% of the tumors with HPV16 accounting for 63.5%, HPV18 for 30.8%, HPV6 for 3.9% and HPV11 for 1.8%. No infection with more than one HPV genotype was detected. HPV infection was significantly associated with poor histological grade, TNM stage I-II, alcohol usage and no smoking status. Multivariate analysis showed that HPV had an independent prognostic effect on the overall survival after adjusting other confounding factors such as histological grade, TNM stage and tobacco usage. The presence of HPV was significantly correlated with a better survival in patients with OSCC.

CONCLUSIONHPV infection can act as an independent predictor for the survival and prognosis of OSCC.

Alcohol Drinking ; Alphapapillomavirus ; classification ; physiology ; Carcinoma, Squamous Cell ; virology ; Cause of Death ; DNA, Viral ; analysis ; Female ; Follow-Up Studies ; Forecasting ; Genotype ; Human papillomavirus 11 ; isolation & purification ; physiology ; Human papillomavirus 16 ; isolation & purification ; physiology ; Human papillomavirus 18 ; isolation & purification ; physiology ; Human papillomavirus 6 ; isolation & purification ; physiology ; Humans ; Male ; Middle Aged ; Mouth Neoplasms ; virology ; Neoplasm Staging ; Papillomavirus Infections ; virology ; Prognosis ; Risk Factors ; Sequence Analysis, DNA ; Smoking ; Survival Rate

The association between cervical cancers and human papillomavirus (HPV) is now well established. To estimate the extent of infection with common HPVs among Korean women, we have examined 224 cervical scrapes of various cervical lesions. Detection and typing of HPVs were done by polymerase chain reaction (PCR) using consensus primers followed by restriction enzyme digestion and PCR using type-specific primers. The prevalence of total HPV infection in patients with cervical intraepithelial neoplasia (CIN) and cervical cancer were significantly higher than those in healthy women and patients with atypical squamous cells of undetermined significance (ASCUS). HPV typing in 41 invasive carcinomas of the cervix revealed the prevalence of HPV 16 in 15 cases, followed by HPV 58, 18, 33, 31, 52 and 35. The distribution pattern of HPV types in CIN were not much different from carcinomas. HPV types except HPV 18 had a tendency to show higher prevalence in high-grade squamous intraepithelial lesion (HSIL) than low-grade squamous intraepithelial lesions (LSIL), however, HPV 18 was detected in LSIL but not in HSIL. HPV 18 tended to have the worse clinical stage, although it was not statistically significant. These findings suggest the importance of HPV typing other than HPV 16 and 18 and a different clinicopathologic significance of HPV 18.
Cervix Neoplasms/virology* ; Cervix Neoplasms/pathology ; Consensus Sequence ; DNA Primers ; DNA, Viral/analysis* ; Female ; Human ; Korea/epidemiology ; Neoplasm Staging ; Papillomavirus, Human/isolation & purification* ; Papillomavirus, Human/genetics ; Papillomavirus, Human/classification ; Papovaviridae Infections/virology ; Papovaviridae Infections/epidemiology* ; Polymerase Chain Reaction/methods ; Polymorphism, Restriction Fragment Length ; Prevalence ; Tumor Virus Infections/virology ; Tumor Virus Infections/epidemiology*

OBJECTIVETo investigate the distribution of HPV 16 variants in Han women patients without Cervical intraepithelial neoplasia in the diagnosis and treatment center for cervical disease, department of Obstetrics and Gynecology in China-Japan friendship hospital with HPV 16 E5 sequence phylogenetic analysis.

METHODSPCR amplification of HPV 16 E5 sequences and sequenced. The association between variations types and different cervical lesions was analyzed.

RESULTSIn this research, We first found that variant classification based on HPV 16 E5 DNA sequence (236 bp) alone had high rate of accuracy. In addition, for the first time, our research revealed that single-4075T can distinguished the As variant from all other variants.

CONCLUSIONIf E5 sequence was used for phylogenetic analysis, it will greatly reduce the experimental costs and improve efficiency and cost-effectiveness. For the first time, our research revealed that single 4075T can distinguished the As variant from all other variants.

Adult ; Cervix Uteri ; virology ; Evolution, Molecular ; Female ; Genetic Variation ; Human papillomavirus 16 ; classification ; genetics ; isolation & purification ; Humans ; Middle Aged ; Molecular Sequence Data ; Oncogene Proteins, Viral ; genetics ; Papillomavirus Infections ; virology ; Phylogeny ; Young Adult

OBJECTIVE: To investigate the correlation of laryngeal squamous cell carcinoma (LSCC) and precancerous lesion with HPV infection subtypes and possible clinical relationship. METHOD: Eighty-three cases in paraffin embedded tissues were detected with thirty seven HPV subtypes by flow-through hybridization and gene chip (HybriMax), including 31 cases of laryngeal squamous cell carcinoma, 52 cases of precancerous lesions (29 cases of vocal cord leukoplakia and 23 cases of laryngeal papilloma), and 36 cases of vocal cord polyp as normal vocal mucosa were used as control. RESULT: The total positive rate of HPV was 19.4% in the group of laryngeal squamous cell carcinoma (6/31), 0 in vocal cord leukoplakia, 65.2% in laryngeal papilloma (15/23), and the control group were all negative, HPV virus subtype of HPV-positive laryngeal squamous cell carcinoma were all high-risk HPV16; and there were 6 HPV virus subtypes in laryngeal papilloma (8: HPV6,4: HPV52, 1: HPV11, 1: HPV18, 2: HPV45, 3: HPV16), individual mixing two or more subtypes infection. HPV infection of laryngeal squamous cell carcinoma and precancerous lesions has no statistically significant difference according to gender, high low-risk subtypes. CONCLUSION HPV infection related to laryngeal squamous cell carcinoma and precancerous lesions, but no significant correlation with the subtype distribution of high and low risk; HPV detection is making positive sense to clinical diagnosis of laryngeal carcinoma and precancerous lesions as well as the development of specific HPV subtype vaccine.
Carcinoma, Squamous Cell ; complications ; virology ; Genotype ; Head and Neck Neoplasms ; complications ; virology ; Human papillomavirus 11 ; Humans ; Laryngeal Neoplasms ; complications ; virology ; Papilloma ; complications ; virology ; Papillomaviridae ; classification ; Papillomavirus Infections ; complications ; virology ; Precancerous Conditions ; Squamous Cell Carcinoma of Head and Neck

OBJECTIVETo study the distribution of human papillomavirus type 16 (HPV16) variants and their clinical significance in Han women with Cervical Intraepithelial Neoplasia (CIN).

METHODSRandomly making a collection of DNA samples of cervical cells from 77 Han out-patients infected with HPV16, PCR amplification of HPV16 DNA fragments containing E6 and E7 genes and sequenced. To study the HPV16 variants types in these out-patients and explore the relationship between the HPV16 variants and CIN by comparing the E6 genes sequenced with the reference strains downloaded from the GenBank. RESULTS Among 77 patients, the minimum age is 21 years old, the maximum age is 56 years old, and the average age is 36.39 +/- 6.86 years old. 61 patients (accounting for 79.2%) were diagnosed as CIN II and higher grade lesions while 16 patients (accounting for 20.8%) as CIN I. In this research, only European variant and Asian variant were found by Parsimony analyses of the sequences. There are 38 Asian variants and 39 European variants. With Chi2 test, Chi2 = 0.0034, P = 0.9535 > 0.05, it suggested that there was no enough evidence to support Asian- and European-variants had the different risk in the cause of cervical intraepithelial neoplasia and cervical cancer.

CONCLUSIONIt was not found Asian- and European-variants of HPV16 had different effect on the cervical cancer, but found only two major variants-Asian- and European-variants in Han people in this research. So we have reason to speculate that there are two major HPV16 variants (Asian- and European-variants) in China's Han women, while other variants, especially high cancer-causing Asian/American variant are not common.

Adult ; Cervical Intraepithelial Neoplasia ; Female ; Genetic Variation ; Human papillomavirus 16 ; classification ; genetics ; isolation & purification ; Humans ; Middle Aged ; Molecular Sequence Data ; Oncogene Proteins, Viral ; genetics ; Papillomavirus E7 Proteins ; genetics ; Papillomavirus Infections ; pathology ; virology ; Phylogeny ; Repressor Proteins ; genetics ; Risk Factors ; Uterine Cervical Neoplasms ; pathology ; virology ; Young Adult