OBJECTIVETo establish a new and rapid GeXP based multiplex PCR assay for the detection and typing of human papillomavirus 6, 11, 31, 33 and 52.

METHODSNucleotide sequences of HPV6, HPV11, HPV31, HPV33 and HPV52 from NCBI were obtained and compared. Genotype-specific primers were then designed and the sensitivity and specificity of multiple PCR assay was evaluated. Optimized assay was further validated with 30 clinical specimens collected from the cervical secretions of patients.

RESULTSA GeXP based multiplex PCR was developed for sensitive detection and reliable differentiation of five HPV genotypes (HPV6, 11, 31, 33 and 52),

CONCLUSIONA GeXP based multiplex PCR assay is demonstrated to be a new and rapid technique for simultaneous detection and typing of 5 different human papillomaviruses.

Adult ; Female ; Humans ; Middle Aged ; Papillomaviridae ; classification ; genetics ; isolation & purification ; Polymerase Chain Reaction ; methods

OBJECTIVETo investigate the prevalence and subtype distribution of human papillomavirus (HPV) infection and its correlation with age among women in Beijing urban area, and provide some epidemiological evidence for the clinical application of HPV vaccines.

METHODSWe collected cervical specimens from 1999 women in the Outpatient Department of our hospital, performed genetyping of HPV-DNA, and analyzed the incidence of HPV infection in different age groups.

RESULTSHPV infection was detected in 502 (25.2%) of the 1999 women patients, with 391 (19.6%) cases of high-risk HPV, which included 326 (83.4%, 326/391) cases of single infection. HPV-16 was the most common type (21.2%, 69/326), followed by HPV-52 (19.3%, 63/326) and HPV-58 (16.0%, 52/326). The prevalence of HPV infection was the highest among the women aged 41 -50 years and the lowest among those over 60 years.

CONCLUSIONThe subtype- and age-specific distribution of HPV infection among women in Beijing urban area shows an obvious heterogeneity, which deserves due consideration in the clinical application of HPV vaccines.

Adolescent ; Adult ; Age Distribution ; Aged ; China ; epidemiology ; Female ; Genotype ; Humans ; Middle Aged ; Papillomaviridae ; classification ; genetics ; Papillomavirus Infections ; epidemiology ; Young Adult

OBJECTIVETo establish a method for rapid detection and sub-typing of human papilloma virus (HPV).

METHODSWe utilized the piezoelectric genosensor (PG) technique, which is a combination of the piezoelectric biosensor and gene chips for HPV identification in 22 recurrent biopsy specimens and 22 corresponding original biopsy specimens. The control samples came from normal tissue of healthy persons. A combined reaction took place on the sensor surface between the target genes and probes. The frequency of the piezoelectric sensor will decrease when such reactions occur, and the frequency decrease depends on the concentration of the target gene. Specimens were also analyzed with conventional PCR and dot blot.

RESULTSOf the 22 recurrent specimens, 15 contained HPV6 DNA, 2 HPV11 DNA, and 4 HPV16 DNA. Only one specimen was negative. All the 22 original specimens were positive: 17 harbored HPV6 DNA, 3 sequence homologous HPV11 DNA, and 2 HPV16 DNA. No HPV18 DNA was detected in any specimen. When compared with PCR and dot blot analysis, the results were essentially the same except for one specimen, which was shown to contain other sub-types of HPV.

CONCLUSIONOur results show that the piezoelectric genosensor technique is a rapid and specific method to analyze HPV.

Biosensing Techniques ; DNA, Viral ; analysis ; Humans ; Papillomaviridae ; classification ; genetics ; Papillomavirus Infections ; virology ; Polymerase Chain Reaction ; Tumor Virus Infections ; virology

OBJECTIVETo investigate the positivity rates and genetic polymorphism of human papillomavirus (HPV) type 81 (HPV81) L1 gene in patients with cervical cancer in Guangdong.

METHODSHPV L1 genes fragment in 1200 positive cervical samples detected with HC-II for high-risk HPV DNA were amplified by HPV-specific PCR with consensus primers, and the L1 genes of HPV81 were sequenced for mutation analysis.

RESULTSFour HPV81-positive cases were found in the total of 1200 cases for the positivity rate of 0.33%. Point mutations of HPV81 L1 gene were found in all of the samples.

CONCLUSIONCompared with the HPV81 prototypes published in GenBank, several new point mutations are found in the L1 genes of the HPV81 in Guangdong, and these mutations are close to those found in Japan.

Adult ; Capsid Proteins ; genetics ; China ; Female ; Humans ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; classification ; genetics ; Papillomavirus Infections ; virology ; Point Mutation ; Polymorphism, Genetic ; Uterine Cervical Neoplasms ; virology

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

OBJECTIVETo detected HPV human papillomavirus DNA and HPV types in NIP nasal inverted papilloma, to inquire into the infection of HPV in the pathogenesis of NIP, and the relationship of HPV with the prognosis of NIP.

METHODSTwenty-eight cases of NIP were divided into 3 groups: no recurrence group (group 1), recurrence group (group 2), squamous cell carcinoma (SCC) arising in NIP group (malignancy group, group 3). Ten cases of benign nasal polyps were used as control group. HPV-DNA types of 6, 11, 16, 18 and general type were detected by polymerase chain reaction (PCR).

RESULTSTotal positive rate of HPV in NIP was 75% (21/28). The positive rate of group 1 was 42% (5/12), all with single and low risk HPV type infection (4 with HPV6 and 1 with HPV11). Thirteen cases of recurrence and 3 cases of malignancy were discovered to have HPV-DNA, in group 2, the majority were HPV6 and HPV11, 4 cases with double infection;in group 3, the majority were HPV16 and HPV18, and 2 cases with double infection.

CONCLUSIONSThe occurrence of NIP was related with HPV infection. To detected HPV and its subtypes can show the cases easily to have recurrence or malignant change. For the cases with HPV positive, double infection and infected with high risk types should be closely followed-up.

Case-Control Studies ; Humans ; Nose Neoplasms ; diagnosis ; virology ; Papilloma, Inverted ; diagnosis ; virology ; Papillomaviridae ; classification ; genetics ; Papillomavirus Infections ; diagnosis ; virology ; Paranasal Sinus Neoplasms ; diagnosis ; virology ; Prognosis

OBJECTIVETo study the circulation, distribution, and genomic diversity of HPVs in common warts in Beijing area of China.

METHODSForty eight patients with pathologically diagnosed common warts were screened for the presence of HPV with HPV type-specific PCR and direct sequencing analysis. The genomic diversity of HPVs prevalent in Chinese patients was analyzed based on LCR.

RESULTSForty one (85.5%) samples were positive for HPV DNA, 13 (31.7%)--HPV-57, 12 (29.3%)--HPV-1a, 7 (17%)--HPV-27 and 5(12.2%)--HPV-2a. Four cases were infected with two different HPV types, two (4.9%) with HPV-1a and HPV-27, one (2.4%) with HPV-1 and HPV-57 and one (2.4%) with HPV-27 and HPV-57. In contrast to the prevalence of single strain of novel HPV-57 variant and HPV-1 prototype, two HPV-2 and three HPV-27 novel variants were found to circulate in Beijing.

CONCLUSIONHPV-1, -2, -27 and -57 are predominantly prevalent in patients with common warts in Beijing.

Adolescent ; Adult ; Aged ; China ; epidemiology ; DNA, Viral ; Female ; Genetic Variation ; Humans ; Male ; Middle Aged ; Papillomaviridae ; classification ; genetics ; isolation & purification ; Phylogeny ; Prevalence ; Warts ; epidemiology ; virology

OBJECTIVEThis study was designed to determine the prevalence of oncogenic human papillomavirus (HPV) in cervical infections in Beijing, China, and to investigate the odds ratio (OR) of HPV single and multiple infections in abnormal cytology.

METHODSA total of 19,018 specimens from outpatients in the department of obstetric and gynecology were collected. They were detected using high-risk HPV genotyping real-time polymerase chain reaction (PCR) kit and analyzed by ThinPrep cytology test for cervical pathological diagnosis. HPV prevalence, age-specific prevalence, and OR of each type of HPV in abnormal cytology were analyzed.

RESULTSOverall, 19.1% (3,623/19,018) of the individuals were positive for HPV infection, 14.9% (2,833/19,018) were positive for a single HPV type, and 4.2% (790/19,018) were positive for multiple types. Among the 3,623 HPV-positive individuals, the most predominant HPV types were HPV52 (4.4%, 834/19,018), HPV16 (3.7%, 710/19,018), and HPV58 (3.4%, 644/19,018). The OR of multiple infections and single infection differed significantly among disease severities. The OR of dual infection was higher than that of each of the two single infection types, respectively.

CONCLUSIONHPV prevalence in the outpatients was 19.1%, and the most predominant HPV types in the study were HPV52, HPV16, and HPV58. Women with multiple infectionswere more likely to have abnormal cytology.

Adolescent ; Adult ; Aged ; Beijing ; Female ; Genotype ; Humans ; Middle Aged ; Papillomaviridae ; classification ; genetics ; isolation & purification ; Papillomavirus Infections ; pathology ; virology ; Uterine Cervical Neoplasms ; pathology ; virology ; Young Adult

We reported a case of an African American woman who went to the hospital with palpable right breast lump with bloody nipple discharge at University of Texas Medical Branch at Galveston. The modalities of breast imagings included mammography and ultrasonography. The method used for viral identification was Linear Array HPV genotyping test. Intraductal papilloma revealed as high density tubular or rounded lobular masses with partially circumscribed, obscured margins and clustered punctate microcalcifications on mammograms. Ultrasound showed as intraductal masses with dilated ducts. The core biopsy demonstrated duct filled with papillary lesion and post excision revealed intraductal papilloma. HPV DNA types 16, 33, 58 and 71 were detected after use of Linear Array HPV genotyping test.
Breast Neoplasms ; diagnostic imaging ; pathology ; virology ; Female ; Genotype ; Humans ; Mammography ; Middle Aged ; Papilloma, Intraductal ; diagnostic imaging ; pathology ; virology ; Papillomaviridae ; classification ; genetics ; isolation & purification

OBJECTIVETo develop a new platform for genotyping human papillomavirus (HPV) and to investigate its effect in clinical application.

METHODSA pair of common primers of 18 HPV subtypes for PCR, was designed in HPV conservative L1 region. Genotyping probes for detecting 15 high-risk HPV subtypes 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66 and 68, together with 3 low-risk HPV 6, 11 and 42 were selected respectively from Genbank and fixed on membrane to make DNA chip. PCR amplification and DNA chip technology were optimized. 100 clinical samples were used to investigate the effect of HPV genotyping DNA chip. Veracity of the genotyping results was verified by sequencing.

RESULTSFrom the 100 clinical samples, 30 were found to be HPV positive, including high-risk HPV subtypes 16, 18, 33, 45, 51, 58, and 66, and low-risk HPV 6, 11 and 42. The sensitivity tested by standard samples was up to 10 copies of HPV DNA.

CONCLUSIONThe HPV genotyping system developed here with DNA chip showed high sensitivity and specificity, suitable to be applied in clinical practice for HPV diagnosis and investigation on the prevalence of HPV sub-types.

DNA Probes, HPV ; genetics ; DNA, Viral ; genetics ; Female ; Genotype ; Humans ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; methods ; Papillomaviridae ; classification ; genetics ; isolation & purification ; Papillomavirus Infections ; diagnosis ; virology ; Polymerase Chain Reaction ; Sensitivity and Specificity ; Uterine Cervical Neoplasms ; virology