Despite being deliberately targeted to common chromosome aneuploidies, the rapid quantitative fluorescent polymerase chain reaction (QF‐PCR) tests can detect the majority of chromosome abnormalities in prenatal diagnosis. The main advantages of this assay are low cost, speed and automation allowing large‐scale application.

Background: Human Papillomavirus (HPV) infection has been well established as etiological agent of cervical cancers. Introduction of HPV genotyping either in cervical screening programs or for monitoring the effectiveness of HPV vaccination, requires access to fast, simple and high-throughput technology.Objectives: Our objective was to evaluate a new multiplex fluorescent PCR method for HPV detection and typing assessing high risk HPV frequency and relationship with cytological and histological data.Materials and Methods: Cervical specimens were collected from 4,051 Italian women (15-65 years aged ) with abnormal cytological smears and were tested for the presence of HPV using the f-HPV typing Kit, a recently available commercial assay (Molgentix Barcelona). Extracted DNA was amplified with 15 sets of fluorescent labelled primers recognising HPV type 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 6 and 11. Labelling of primers with 6-FAM, NED, VIC and PET allowed high sensitivity as well as amplification of similar size products in the same PCR reaction. After electrophoresis on ABI 3130 DNA sequencer, PCR products were automatically recognised by size and colours. Cytological and histological analysis was performed.Results: High risk HPV types were detected in 80% (1,145/4,091) and low risk HPV in 20% (284/4,091) of the samples. High risk HPV were detected in 42% (43/102) of ASCUS-AGUS, 50% (234/466) of CIN1, 70% (88/126) of CIN2 , 97% (35/36) CIN3, 98% (118/120) of squamous cervical cancer and finally in 100% (25/25) of adenocarcinomas. HPV-16 was the most prevalent type in LSILs (40%), in HSILs (72%), and in carcinomas (62%) followed by HPV-31.Conclusions: The f-HPV typing™ kit is an accurate technique for quick and sensitive detection of 15 HPV types, it also allows distinguishing single and multiple infections in several cases. The high throughput of the assay allows assessing different HPV types frequency in a population and may be of help in determining possible changes that could derive as consequence of vaccination policies.

Objective: Detection of high-risk human papillomavirus types (HPV) infection is an important tool in the screening of cervical cancer and triage of cytological abnormalities. The different techniques for detection of this cancer need to be contrasted and validated for use in population screening. Material and Methods: Cervical cell samples were collected from 166 women attending a dermatology clinic in Oviedo (Spain).We evaluated the performance of three different assays for VPH detection.The methods utilized were 1) In-house PCR-EIA using LI consensus primers MY09/ MY 11, 2) A PCR-reverse line blot hybridization (PCR-LBH) that uses L I consensus PGMY primers. 3) Hybrid Capture 2.All assays were performed blinded.The kappa statistic was used to test for global agreement between assay pairs. Results: HPV DNA was detected in 24,7%, 25,3% and 29,5% of the women, respective to the assay. The overall agreement between the in-house PCR, PCR-LBH and HC2 was (73.5%) with all kappa values between assay pairs exceeding 0.56 (p < 0.001). Conclusion:The three HPV assays were equally accurate in estimating high-risk HPV prevalence and HPV-related lesions.The method for HPV detection must be decided depending on the goals of the search (screening, follow-up or molecular studies).

OBJETIVO: La identificación de la infección por tipos de alto riesgo del virus del papiloma humano (VPH) es una herramienta útil para el cribado de cáncer del cuello uterino. Las distintas técnicas aplicadas para su detección deben contrastarse y validarse para su empleo en la tamización poblacional. MATERIAL Y MÉTODOS: Se evalúan tres técnicas para la detección del VPH en 166 muestras cervicales procedentes de mujeres atendidas en una clínica de dermatología en Oviedo (España): a) PCR-EIA mediante consensos MY09/MY011; b) PCR con line blot hybridization (PCR-LBH) con consensos PGMY; y c) hybrid capture 2. RESULTADOS: El ADN-VPH se reconoció en 29.5%, 25.3% y 24.7%, de acuerdo con el ensayo. La concordancia global entre PCR-EIA, PCR-LBH y HC2 fue de 73.5% con los valores de kappa superiores a 0.56 entre los ensayos (p<0.001). CONCLUSIONES: La prevalencia de tipos de alto riesgo oncogénico así como de las lesiones fue similar en los tres ensayos. En virtud de que las técnicas son comparables, su elección debe basarse en las condiciones individuales de cada laboratorio y el volumen de muestras por procesar.OBJECTIVE: Detection of high-risk human papillomavirus types (HPV) infection is an important tool in the screening of cervical cancer and triage of cytological abnormalities. The different techniques for detection of this cancer need to be contrasted and validated for use in population screening. MATERIAL AND METHODS: Cervical cell samples were collected from 166 women attending a dermatology clinic in Oviedo (Spain). We evaluated the performance of three different assays for VPH detection. The methods utilized were 1) In-house PCR-EIA using L1 consensus primers MY09/MY11, 2) A PCR-reverse line blot hybridization (PCR-LBH) that uses L1 consensus PGMY primers. 3) Hybrid Capture 2. All assays were performed blinded. The kappa statistic was used to test for global agreement between assay pairs. RESULTS: HPV DNA was detected in 24,7%, 25,3% and 29,5% of the women, respective to the assay. The overall agreement between the in-house PCR, PCR-LBH and HC2 was (73.5%) with all kappa values between assay pairs exceeding 0.56 (p<0.001). CONCLUSION: The three HPV assays were equally accurate in estimating high-risk HPV prevalence and HPV-related lesions. The method for HPV detection must be decided depending on the goals of the search (screening, follow-up or molecular studies).

Detection of high-risk human papillomavirus types (HPV) infection is an important tool in the screening of cervical cancer and triage of cytological abnormalities. The different techniques for detection of this cancer need to be contrasted and validated for use in population screening.Cervical cell samples were collected from 166 women attending a dermatology clinic in Oviedo (Spain). We evaluated the performance of three different assays for VPH detection. The methods utilized were 1) In-house PCR-EIA using LI consensus primers MY09/ MY11, 2) A PCR-reverse line blot hybridization (PCR-LBH) that uses LI consensus PGMY primers. 3) Hybrid Capture 2. All assays were performed blinded. The kappa statistic was used to test for global agreement between assay pairs.HPV DNA was detected in 24,7%, 25,3% and 29,5% of the women, respective to the assay. The overall agreement between the in-house PCR, PCR-LBH and HC2 was (73.5%) with all kappa values between assay pairs exceeding 0.56 (p<0.001).The three HPV assays were equally accurate in estimating high-risk HPV prevalence and HPV-related lesions. The method for HPV detection must be decided depending on the goals of the search (screening, follow-up or molecular studies).