Multiplex PCR 기법을 이용한 보통사마귀 내 인유두종바이러스 검출 및 분류

HPV Genotype Detection Using Hybrid Capture Sample Preparation Combined with Whole Genome Amplification and Multiplex Detection with Luminex XMAP

Objective: The aim of this study was to investigate the relationship between human papilloma virus (HPV) types which was worked with polymerase chain reaction (PCR) and immunohistochemistry methods and cvtology and biyopsy results. Materials and Methods: A total of 100 cases from hospital between 2010-2013 participitated in this study. Linear Array HPV genotyping test was worked with PCR, tissue biopsies was evaluated with light microscopy, HPV was detected with immunohistochemistry (IHK) method and results were compared evaluating of conventional smear preparations according to Bethesda. Results: While 61 cases included high risk (61%), 39 cases included low risk HPV (39%). HPV types with high risk such as HPV16, HPV6, HPV11, HPV18, HPV31 respectively detected in 24, 19, 9, 7, 2 cases. HPV types with low risk such as HPV66 detected in 7 cases. HPV16 was observed epithelial cell abnormality such as 9 ASCUS, 3 LSIL,1 ASC-H, 1 AGUS in 14/24 ratio (58%) and ILMAN was observed 10/24 ratio (42%). Between cytological diagnosis results and HPV types are significant relationship according to chi-square test (p=0.001). When the results of the biopsy tissue analyzed, 38 patients were diagnosed intraepithelial neoplasia and malignant, 62 cases were diagnosed inflammation and other benign featured. The relationship between tissue diagnosis and HPV types is significant according to chi-square test (p<0.001). HPV in tissue was positive in 44 cases. The relationship between HPV types and tissue HPV results are significant (p<0.001). The cytological tissue diagnosis of 19 patients with ASCUS has been reported as follows; 10 CIN1, 3 CIN2, 2 CIN3 and 4 inflammation. Negative and positive HPV results studied in tissue are not significant results with cytological diagnosis (p=0.02) and tissue diagnosis results (p=0.02). Conclusion: The important of HPV detection is obvious, but tissue changes as well as HPV detection are important. Thus combined use of testing, even in evaluation with the biopsy results may give more realistic results for all types of cancers worldwide.

Verruca vulgaris (VV) is a prevalent skin condition caused by various subtypes of human papilloma virus (HPV). The most common causes of non-genital lesions are HPV types 2 and 4, and to a lesser extent types 1, 3, 26, 29, and 57. Although numerous therapeutic modalities exist, none is universally effective or without adverse events (AE). Pulsed dye laser (PDL) is a favorable option due to its observed efficacy and relatively low AE rate. However, it is not known which verrucae are most likely to respond to PDL, or whether the causative viral subtype influences this response. The objective of this prospective blinded study was to assess whether the HPV subtype was predictive of response to PDL. For that matter, 26 verrucae from 26 immunocompetent patients were biopsied prior to treatment by PDL. HPV coding sequences were isolated and genotyped using PCR analysis. Patients were treated by PDL (595nm wavelength, 5mm spot size, 1.5ms pulse duration, 12J/cm2 fluence) once a month for up to 6months, and clinical response was assessed. Binary logistic regression analysis and linear logistic regression analysis were used in order to evaluate statistical significance. Different types of HPV were identified in 22 of 26 tissue samples. Response to treatment did not correlate with HPV type, age, or gender. As no association between HPV type and response to PDL therapy could be established, it is therefore equally effective for all HPV types and remains a favorable treatment option for all VV.

Aim. Toassess the prevalence of various types of human papillomavirus (HPV) of high carcinogenic risk (HCR) and the rate of abnormal findings of cytological examination in HPV-positive and HPV-negative women of the Kemerovo region.&#x0D; Materials and methods. A continuous cross-sectional study was conducted. The study included 675 women aged 21 to 65 (mean age 40.510.6 years) residing in the Kemerovo region. Patients were sampled from the cervical canal for semi-quantitative HPV-typing and cytological examination with Papanicolaou staining (PAP-test) using a liquid-based cytology method. For HPV-positive patients, an HPV typing with the quantification of 14 HCR HPV types was performed by polymerase chain reaction with real-time detection. Statistical data processing was performed using the StatSoft Statistica 6.1 software bundle.&#x0D; Results. The mean rate of HCR HPV detection in women of the Kemerovo region in the age group of 2165 years is 17.8% and decreases with age. HPV type 16 prevailed and was detected in 31.6% of women. In 19.2% of women, the HPV subtype 31 was diagnosed; in 18.3%, the subtype 35; in 16.7%, the subtype 33. The HPV subtypes 39, 56, and 66 were the rarest. One subtype of HCR HPV was detected in 26.7% of women, two subtypes in 6.7%, and three or more subtypes in 66.6% of women. According to the cytological examination, CIN 1, 2, and 3 rates were 3.3, 0.8, and 0.7%, respectively. HSIL (CIN 23) was detected in 1.5% of cases, LSIL (CIN 1) in 3.3%. The rate of CIN 1 in HPV(+) women was higher: 13.3% vs 1.0% in HPV(-) (2=43.197; p0.001). CIN 2 and CIN 3 were reported in HPV(+) patients only. HSIL+ was diagnosed in 7.9% of HPV(+) females (2=39.778; p0.001).&#x0D; Conclusion. Thus, the study results showed that the mean infection rate of HCR HPV in women of the Kemerovo region at the age of 2165 years is 17.8% and decreases with age. HPV subtype 16 is predominant; subtypes 31, 35, and 33 were less common. In HPV-positive women, abnormal findings of cytological examination were significantly more common.

Human papillomavirus type 6 and 11 and head and neck squamous cell carcinomas in sudanese patients

AIM: To evaluate the presence of high risk human papillomaviruses (HPV) in cervical smears showing intraepithelial neoplasia (CIN). METHODS: The presence of 14 high risk HPV was evaluated in 114...

Cutaneous warts are caused by human papillomavirus (HPV). To date, more than 120 different types of HPV are known, of which 80 have been completely characterized. Prevalence studies on types of HPV present in cutaneous warts have been carried out in immunocompetent individuals and immunosuppressed organ allograft recipients, but not in human immunodeficiency virus (HIV)-positive patients. To determine the HPV types present in cutaneous warts of HIV-infected patients. Twenty-five biopsies of cutaneous warts from HIV-infected patients and 14 samples from control non-HIV-infected patients were studied. HPV detection was performed by polymerase chain reaction using two sets of primers: MY09/MY11 and RK91. The type of HPV was determined by restriction fragment length polymorphism analysis and direct sequencing of the amplified products. HPV DNA was detected in 64% of cutaneous warts from HIV-infected patients and in 79% of samples from the control group. The HPV types identified in HIV-infected patients were: HPV 2 (38%), 57 (31%), 27 (12%), 6 (12%) and 7 (6%). HPV 2/27/57 predominated in both groups, being present in 81% of lesions from HIV-infected patients and 82% of samples from non-HIV-infected patients. HPV 6, a genital HPV type rarely found in cutaneous lesions, was detected in two warts from HIV-infected patients and in one lesion of the immunocompetent group. HPV 7, characteristically associated with butcher's warts, and recently detected in oral and perioral lesions of HIV-infected patients, was found for the first time in a non-facial lesion of an HIV-infected patient. This is the first study evaluating the prevalence of HPV types in cutaneous warts of HIV-infected patients and immunocompetent individuals in Brazil.

To study the distribution of human papilloma virus (HPV) subtypes in Uyghur people from the high risk region of cervical cancer in Xinjiang. 21 subtypes of HPV from 400 cases of men's swabs of penile and 400 cases of women's cervix swabs were detected, using flow-through hybridization and gene chip technology. (1) The positive rate of HPV among the 400 women cases was 14.25%, with HPV16 the most common type. In women whose HPV was positive, the positive rate of HPV16 was 66.67%. The ranking of frequencies on the HPV subtypes in women were:HPV16, 58, 39, 18, 33, 52, 43, 66, CP8304, HPV6 and 11. (2) Among the 400 male cases, the positive rate of HPV was 8.00%, with HPV16 the most common type. In men whose HPV was positive, the rate of HPV16 was 44.44%. The ranking of HPV subtypes in men were HPV16, 43, 33, 39, 6 (HPV39 and 6 are equivalent) and 53. (3) Both wives and husbands were infected by different subtypes of HPV, the positive rate was relatively low among men whose wife's HPV were positive. However, the HPV positive rate was relatively low among women whose husband's HPV were positive. The concordance infection rate of women and men were 7.02% and the concordance infection rate of men and women were 9.38%. The HPV positive rates were relatively low in both men and women living in the region with high risk of cervical cancer in Xinjiang. However, the positive rate of high risk types HPV and HPV16 in both men and women were relatively high. Uyghur men seemed to have played a certain role related to the carcinogenesis of cervical cancer. HPV16 was the main type in both men and women in Xinjiang.

Estimates of transmissibility of infectious agents such as human papillomavirus (HPV) are important for developing preventive measures and determining the impact of such interventions. Based on the results of epidemiologic studies, HPV appears to be highly transmissible;1–8 however, empirical data on HPV transmissibility are lacking.9–11 We sought to design and conduct a study among couples to provide the first empirical estimate of male-to-female sexual transmissibility of HPV. However, we faced several difficulties in achieving this goal. Here we present the design and descriptive results of this study in brief and these difficulties in detail. Information on study population arose from a larger study described in detail elsewhere.5 Briefly, between December 2000 and November 2006, 244 women (age range: 18–22) were recruited to a longitudinal study of HPV infection through invitational letters mailed to 24,201 female University of Washington undergraduates and informational flyers at the student health clinic. Participants were eligible to enroll if they had never had vaginal intercourse with a male partner (i.e., virgin) or had first had intercourse with one male partner within the previous three months (i.e., peri-virgin). Consenting women were asked to refer their first male partner to the study and were given a letter of recruitment to forward to him. Every two weeks, women completed a Web-based diary designed to capture daily sexual behavior information such as date and number of acts of intercourse with a new partner.5, 12 Women were followed-up with gynecological examinations every four months. At each visit, self-collected vaginal swabs and clinician-collected cervical and vulvovaginal swabs were collected. Men were seen only for one visit. They underwent a clinician-administered interview about their medical and sexual history and a genital examination including specimen collection from the penis, urethra, and scrotal area, described in detail elsewhere.13 All samples were tested for HPV DNA by polymerase chain reaction (PCR)-based methods. Specimens testing positive by a generic probe were typed using a reverse line-blot assay (Roche Molecular Systems) for 37 distinct HPV types.14 PCR results of vaginal, cervical, and vulvovaginal swabs for women and penile, urethral, and scrotal swabs for men were combined to determine genital HPV status in each participant. Institutional Review Board of the University of Washington approved the study; informed consent was obtained from all participants. Male partners were enrolled for 33 women. Information on 19 couples was excluded due to the following reasons: one woman did not report any sexual activity in the diary; three men were not the first sex partner of their female partner; and 15 couples were not found to be infected with HPV. Of the remaining 14 couples, both partners were found to be infected with HPV in five couples and the male partner only was found to be infected in three couples. Descriptive results for these eight couples and for the other six couples in whom, unexpectedly, the female partner was found to be infected with HPV, while the male partner was not, are provided in Tables 1 and ​and22. Table 1 Characteristics of eight couples in whom at least one type of HPV was detected in the single visit of the first male partner Table 2 Characteristics of six couples in whom at least one type of HPV was detected in the female partner but not in the male partner Investigators who embark upon designing a methodologically sound study to find empirical estimates of male-to-female sexual transmissibility of HPV may face the following difficulties:

BackgroundAlthough high risk HPVs are associated with an increased risk of prostate cancer it is not known if they have a causal role. The purpose of this study is to investigate the potential role of human papilloma viruses (HPVs) in prostate cancer. The aims are (i) to investigate the presence and confirm the identity of high risk HPVs in benign prostate tissues prior to the development of HPV positive prostate cancer in the same patients, and (ii) to determine if HPVs are biologically active.MethodsWe used polymerase chain reaction (PCR) to identify HPVs in specimens from 52 Australian men with benign prostate biopsies who 1 to 10 years later developed prostate cancer. Immunohistochemistry (IHC) was used to assess the expression of HPV E7 oncoproteins, cytokeratin and prostate specific antigen (PSA).We used RNASeq data from The Cancer Genome Atlas (TCGA) to identify possible HPV RNA sequences in prostate cancer.ResultsHPV screening using standard PCR was conducted on 28 of the 52 sets of benign and later prostate cancers. HPV L1 genes were identified in 13 (46%) benign and 8 (29%) of 28 later prostate cancers in the same patients. HPV E7 genes were identified in 23 (82%) benign and 19 (68%) of 28 subsequent prostate cancers in the same patients. The same HPV types were present in both the benign and subsequent prostate cancers in 9 sets of specimens. HPV type 16 was identified in 15% of benign and 3% of prostate cancers. HPV type 18 was identified in 26% of benign and 16% of prostate cancers. Small numbers of HPV types 45, 47, 76 and 115 were also identified.High confidence RNA-Seq evidence for high risk HPV types 16 and 18 was identified in 12 (2%) of the 502 TCGA prostate cancer transcriptomes.High risk HPV E7 oncoprotein was positively expressed in 23 (82%) of 28 benign prostate specimens but only in 8 (29%) of 28 of the later prostate cancer specimens. This difference is statistically significant (p = 0.001). Prostate specific antigen (PSA) was more highly expressed in 26 (50%) of 52 prostate cancer specimens as compared to prior benign prostate specimens in the same patients.ConclusionsHigh risk HPVs are present in benign prostate tissues prior to the development of HPV positive prostate cancer. There is a significantly higher expression of HPV E7 oncoproteins in benign prostate tissues as compared to late prostate cancer that subsequently developed in the same patients. This observation suggests that HPV oncogenic activity is an early phenomenon in a majority of prostate oncogenesis. TCGA RNA-Seq data suggests that HPV is biologically active in some prostate tumour samples.

<h3>Introduction/Background*</h3> Cervical cancer is the fourth most common cancer, worldwide. Persistent infection with high-risk (HR) human papillomavirus (HPV) types is necessary for cervical cancer development. However, little is known about the influence of multiple HPV infections on cervical lesion risk. The aim of this study was to see the prevalence and pattern of multiple HPV infections in cervical cancer patients from Bangladesh. <h3>Methodology</h3> Histopahologically diagnosed 100 cervical cancer patients were enrolled in this study. HPV DNA testing was done by polymerase chain reaction (PCR) using SPF-10 broad-spectrum primers followed by genotyping by reverse hybridization using the INNO-LIPA genotyping Extra (Fujirebio, Belgium) at the Department of Virology, Bangabandhu Sheikh Mujib Medical University. <h3>Result(s)*</h3> 22.0% of cervical cancer patients were associated with multiple HPV infections whereas overall prevalence of single HPV infections was 78%. Overall, 11 different HPV types [9 HR, 2 intermediate risk (IR) and 1 low risk (LR)] were detected in this study. Among them, the most prevalent genotype was HPV 16, followed by HPV 18,45,56,58,39,31,73,53,66,62. Among the single infection HPV 16 was more prevalent (69%) followed by HPV 18 (6%) and HPV 45 (3%), later eight genotypes were found only in multiple infections. Genotype 16 and 18 alone and as co-infection were detected in 84% cases. Among the multiple infections the most common was by HR-HR HPV types (77.27%), whereas infections by HR-IR and HR-LR HPV types were identified in 13.64% and 9.09%, respectively. Dual, triple and quadruple infection were observed in this study. Overall prevalence of HPV dual infection (16%) was more in multiple infection. Genotypes 16/18 co-infection was more among the dual infection. Among the triple and quadruple infection 16/45/56, 16/18/53/73, 16/18/45/66, and 16/31/56/58 were prevalent. <h3>Conclusion*</h3> Thus this study concluded that HPV multiple infection (22%) is prevalent along with the single HPV infection among the cervical cancer patients from Bangladesh. Dual, triple and quadruple infection were observed in this study. Genotype 16 and 18 alone and as co-infection were detected in 84% cases. Thus the vaccination with Cervarix against HPV 16 and HPV 18 can prevent 84% of cervical cancer in Bangladesh.

Human Papillomavirus Genotypes and Anal-Related Lesions Among HIV-1-Infected Men in Milan, Italy

Background: Multiplex polymerase chain reaction allows amplification of multiple target sequences of a genome under identical conditions in a single tube. This one-shot polymerase chain reaction detection is time and cost effective when large or multiple genes, with many target fragments are investigated. This is applicable for retinoblastoma susceptibility gene having 27 exons with recurrent mutations reported at most of the 12 CGA codons. Materials and Methods: Multiplex polymerase chain reaction assay for the amplification of 12 CGA codons, which constitutes about 50 % of retinoblastoma susceptibility gene mutations has been designed. The time and cost (includes only reagent cost) involved in both multiplex and uniplex polymerase chain reaction was also calculated. Results: Twelve CGA codons were multiplexed in 5 instead of 12 uniplex polymerase chain reactions, which took 36 hours and 9.78 US$ whereas multiplex polymerase chain reaction took 15 hours and 6.88 US$. Multiplex polymerase chain reaction method saved 58.3% of time and 29.6% of cost over uniplex polymerase chain reaction. Conclusion: Saving time by more than half and cost by nearly a third would help clinicians and geneticists while counseling retinoblastoma patients.

The aim of this study was to determine the proportion of invasive cervical cancers attributable to human papillomavirus (HPV) types 16 and 18 in a contemporary, cytologically well-screened UK population. This was achieved in a retrospective observational analysis by HPV typing 453 archival invasive cervical cancers diagnosed between January 1, 2000 and September 1, 2006. Pathological material was collected from 9 hospitals across Wales (UK), and HPV typing and pathology review was conducted at a central laboratory. Genotyping for high-risk HPV DNA was performed by PCR-enzyme immunoassay using the GP5+/6+ primer set. DNA was successfully extracted from 297 cases. Two hundred and eighty cases were included in the final analysis. The proportion of cases which had only HPV 16 and/or 18 was 219 of 280 (78.2%, 95% CI = 73.0-82.7); the proportion of cases which had HPV 16 or 18 and another HPV type was 230 of 280 (82.1%, 95% CI = 77.2-86.2). The proportion of cervical cancers associated with infection with HPV types 16 and 18 has previously been estimated at around 70%. The appropriate figure for a cytologically well-screened UK population appears to be approximately 80%. Hence, the potential impact of the current vaccination programme may be underestimated.

Half of the Nobel Prize in Physiology and Medicine for 2008 has been awarded to Harald zur Hausen, a German virologist, for “his discovery of human papilloma viruses (HPV) causing...