Comparative Evaluation of the HPV28 Detection and HPV DNA Chip Test for Detecting and Genotyping Human Papillomaviruses

BACKGROUND: The HPV28 Detection test (Seegene) is a real-time polymerase chain reaction assay that is designed for testing a total of 28 human papillomavirus (HPV) genotypes and estimating the approximate HPV viral load. The aim of this study was to evaluate the clinical applicability of the HPV28 Detection test with regard to the prevalence of HPV infection and distribution of HPV genotypes by using the HPV28 Detection and HPV DNA Chip tests (Biomedlab). METHODS: HPV DNA Chip and HPV28 Detection tests were performed for 500 cervical swab specimens. HPV genotype results were confirmed by sequencing analysis of the specimens that showed discordant results in the 2 test methods. RESULTS: The positive rate of HPV detection determined by using HPV28 Detection and HPV DNA Chip tests were 43.8% and 40.6%, respectively. The sequencing results in 64 discordant specimens that showed single HPV infection in the 2 test methods were in complete agreement with the test results obtained with the HPV28 Detection test. The genotyping results of the HPV28 Detection test were 100% concordant in repeated experiments with HPV-infected specimens that have 12 different HPV genotypes, i.e., types 16, 31, 33, 39, 42, 51, 52, 53, 58, 66, 68, and 70. The HPV28 Detection test was 100-fold more sensitive than the HPV DNA Chip test with serially diluted HPV DNAs. CONCLUSIONS: The HPV28 Detection test can be applied in the clinical field as an HPV genotyping test can accurately identify various HPV genotypes with high specificity and low detection limit.

Prevalence and Genotype of Human Papillomavirus Infection and Risk of Cervical Dysplasia among Asymptomatic Korean Women / 대한임상미생물학회지

BACKGROUND: The persistence of infection by high-risk human papillomavirus (HPV) may lead to cervical cancer. Recently, the American Society for Colposcopy and Cervical Pathology (ASCCP) announced that oncogenic HPV screening and the PAP smear are the main methods of screening for cervical cancer. The goal of this study was to investigate the prevalence and genotyping of HPV, as well as the risk of cervical dysplasia. METHODS: HPV genotyping was conducted by a commercial chip assay. Cervical dysplasia was retrospectively reviewed using electronic medical records. The study participants were grouped together according to cervical dysplasia status: 'no dysplasia,' 'atypical squamous cells of undetermined significance (ASCUS),' 'low-grade squamous intraepithelial lesion (LSIL),' and 'high-grade squamous intraepithelial lesion (HSIL).' The HPV prevalence and genotyping were analyzed according to the cervical dysplasia group. RESULTS: The overall prevalence of HPV was 17.6% (91 out of 518 patients). HPV-18 (2.3%), HPV-16 (2.1%), and HPV-58 (1.2%) were the three most frequent genotypes. The prevalence of HPV infection and the high-risk HPV positive rate was higher in the ASCUS, LSIL, and HSIL groups than in the no dysplasia group (P<0.05). CONCLUSION: In this study, basic data regarding the prevalence and distribution of HPV genotypes were obtained. Since HPV vaccination has been actively encouraged among Korean women, a change in the prevalence of HPV and cervical dysplasia is expected in the future. This study provided basic data describing the prevalence of HPV and its genotypes in the pre-HPV vaccination era.

Clinical Effectiveness of Restriction Fragment Mass Polymorphism Assay for Human Papillomavirus Genotyping / 임상검사와정도관리

BACKGROUND: Human papillomavirus (HPV) infection is the main cause of cervical cancer and with the advent of genotype specific vaccines, there is increased need for accurate, broad-spectrum and high-throughput methods for HPV genotyping. A MALDI-TOF mass spectrometry (MS)-based restriction fragment mass polymorphism (RFMP) assay has proven to accurately and reliably genotype a wide variety of HPV. METHODS: We evaluated the clinical utility of the RFMP assay in HPV genotyping by testing a total of 2,689 specimens taken from liquid-based cytology, which was composed of normal cytology, atypical squamous cells of undetermined significance (ASCUS), low grade squamous intraepithelial lesion (LSIL), high grade squamous intraepithelial lesion (HSIL) and invasive squamous cervical cancer (SCC). RESULTS: Overall HPV positive rate of total specimens was 32.5% and the high-risk positivity was 16.4%. The HPV positive rates were increased as increasing severity level of cervical lesion. Predominant high-risk HPV genotypes were found as following order; 52 (18.6%), 16 (13.7%), 18 (3.8%), 58 (3.4%), 56 (2.6%) and 31 (2.5%). The high-risk HPV positivities according to cytologic diagnosis were 10.7% (238/2229), 31.7% (76/240), 50.0% (88/176), 86.0% (37/43), 100% (1/1) in normal, ASCUS, LSIL, HSIL and SCC subgroups, respectively. The concordance rate and Kappa value between sequencing and RFMP assays were 96.6% and 0.932 (95%CI: 0.908-0.956). CONCLUSIONS: The RFMP HPV genotyping assays showed high concordance with sequencing. The assay is simple, and can accurately detect and identify HPV genotypes in samples with various levels of cytological lesions. The results demonstrated that RFMP assay should be clinically suitable for HPV genotyping in laboratories.

Human papillomavirus in oral lesions / Virus papiloma humano en lesiones orales

Growing evidence suggests a role for human papillomavirus (HPV) in oral cancer; however its involvement is still controversial. This study evaluates the frequency of HPV DNA in a variety of oral lesions in patients from Argentina. A total of 77 oral tissue samples from 66 patients were selected (cases); the clinical-histopathological diagnoses corresponded to: 11 HPV- associated benign lesions, 8 non-HPV associated benign lesions, 33 premalignant lesions and 25 cancers. Sixty exfoliated cell samples from normal oral mucosa were used as controls. HPV detection and typing were performed by polymerase chain reaction (PCR) using primers MY09, 11, combined with RFLP or alternatively PCR using primers GP5+, 6+ combined with dot blot hybridization. HPV was detected in 91.0% of HPV- associated benign lesions, 14.3% of non-HPV associated benign lesions, 51.5% of preneoplasias and 60.0% of cancers. No control sample tested HPV positive. In benign HPV- associated lesions, 30.0% of HPV positive samples harbored high-risk types, while in preneoplastic lesions the value rose to 59.9%. In cancer lesions, HPV detection in verrucous carcinoma was 88.9% and in squamous cell carcinoma 43.8%, with high-risk type rates of 75.5% and 85.6%, respectively. The high HPV frequency detected in preneoplastic and neoplastic lesions supports an HPV etiological role in at least a subset of oral cancers.

Crecientes evidencias sugieren que el virus Papiloma humano (HPV) tiene un rol en el cáncer oral; sin embargo su participación es todavía controvertida. Este estudio evalúa la frecuencia de ADN de HPV en una variedad de lesiones orales de pacientes de Argentina. Se seleccionaron 77 muestras de tejido oral de 66 pacientes (casos); el diagnóstico histo-patológico correspondió a: 11 lesiones benignas asociadas a HPV, 8 lesiones benignas no asociadas a HPV, 33 lesiones premalignas y 25 cánceres. Como controles se usaron 60 muestras de células exfoliadas de mucosa oral normal. La detección y tipificación de HPV se realizó por PCR empleando los primers MY09,11, seguida de RFLP, o PCR usando los primers GP5+, 6+ seguida de hibridación en dot blot. HPV fue detectado en 91% de las lesiones benignas asociadas a HPV, 14.3% de las lesiones benignas no asociadas, 51.5% de preneoplasias y 60% de cánceres. Ninguna muestra control resultó HPV positiva. En las lesiones benignas, 30% de las muestras HPV positivas correspondieron a tipos de alto riesgo, mientras que en las lesiones preneoplásicas la positividad ascendió a 59.9%. En cánceres, la detección de HPV en carcinomas verrugosos fue 88.9% y en carcinomas escamosos 43.8%, con 75.5% y 85.6% de tipos virales de alto riesgo, respectivamente. La alta frecuencia de HPV detectada en lesiones preneoplásicas y cánceres apoya un rol etiológico del HPV en, al menos, un subgrupo de cánceres orales.

Human Papilloma Virus Genotyping Assay using Restriction Fragment Mass Polymorphism Analysis, and Its Comparison with Sequencing and Hybrid Capture Assays / 대한진단검사의학회지

BACKGROUND: Infection with human papilloma virus (HPV) is the main cause of cervical cancer, and HPV genotyping is of increasing importance for determining clinical course and management of the disease based on the HPV genotypes. Here, we established a novel matrix-assisted laser desorption/ ionization time of flight mass spectrometry (MALDI-TOF MS) assay, termed restriction fragment mass polymorphism (RFMP) that is suitable for genotyping multiple HPV in an accurate and high-throughput manner. We evaluated the performance of the RFMP assay in HPV genotyping by comparing the results with those of direct or clonal sequencing and hybrid capture (HC) assays. METHODS: The study population consisted of 50 patients with histologically confirmed cervical lesions and a positive test for HPV DNA. HPV genotyping was performed with RFMP, sequencing, and HC assays. The assigned genotypes and risk groups were compared among the methods. RESULTS: Concordance rates in the genotype level between RFMP vs sequencing, sequencing vs HC, and HC vs RFMP were 98% (49/50), 88% (44/50), and 88% (44/50), respectivley. Especially, RFMP and sequencing were 100% concordant when assigned high-risk group was considered identical in 1 case of mixed genotypes identified only in RFMP. The observed discrepancy between HC and the other two methods is due to the assignment of six cases of low, intermediate, or unassigned risk genotypes as high-risk group in HC method. CONCLUSIONS: RFMP, sequencing, and HC assays were highly concordant with each other in HPV genotyping. Compared to sequencing assay, RFMP assay is found to be advantageous in detecting mixed genotype infections. The accuracy and amenability to high-throughput analysis should make the RFMP assay suitable for reliable screening of HPV genotypes in clinical laboratories.

Identification of High Risk Group of HPV associated with Cervical Cancer in Korean Women / 대한산부인과학회잡지

OBJECTIVE: To identify genital HPV types and high risk group of HPV associated with cervical cancer in Korean women. METHODS: Both Pap test and HPV-DNA test using PCR assay were performed as screening test for cervical cancer in this clinic. When patients were positive in HPV-DNA test, HPV genotyping using sequencing method and cervical biopsy were performed. RESULTS: Frequent age group of HPV infection was 40 yrs (34.3%) and prevalence of HPV infection was 9.8%. Twenty-three types of HPV were detected. HPV 16 and 58 were detected in invasive cancer. HPV 16, 31, 33, 45, and 58 were detected in HSIL. HPV 6, 11, 18, 53, 59, and 66 were detected in LSIL. HPV 16 was most commonly detected in HSIL and invasive cancer. CONCLUSION: HPV 16, 31, 33, 45, and 58 are included in high risk group of HPV in Korean women. It may be very effective in early detection of cervical cancer to classify HPV types included in high risk group of cervical cancer in Korean women and to perform cervical biopsy in the patients who have high risk types of HPV infection.

Identification of High Risk Group of HPV associated with Cervical Cancer in Korean Women / 대한산부인과학회잡지

OBJECTIVE: To identify genital HPV types and high risk group of HPV associated with cervical cancer in Korean women. METHODS: Both Pap test and HPV-DNA test using PCR assay were performed as screening test for cervical cancer in this clinic. When patients were positive in HPV-DNA test, HPV genotyping using sequencing method and cervical biopsy were performed. RESULTS: Frequent age group of HPV infection was 40 yrs (34.3%) and prevalence of HPV infection was 9.8%. Twenty-three types of HPV were detected. HPV 16 and 58 were detected in invasive cancer. HPV 16, 31, 33, 45, and 58 were detected in HSIL. HPV 6, 11, 18, 53, 59, and 66 were detected in LSIL. HPV 16 was most commonly detected in HSIL and invasive cancer. CONCLUSION: HPV 16, 31, 33, 45, and 58 are included in high risk group of HPV in Korean women. It may be very effective in early detection of cervical cancer to classify HPV types included in high risk group of cervical cancer in Korean women and to perform cervical biopsy in the patients who have high risk types of HPV infection.