PCR-RFLP assay as an option for primary HPV test

Abstract

Persistent human papillomavirus (HPV) infection is an essential factor of cervical cancer. This study evaluated the analytical performance of restriction fragment length polymorphism polymerase chain reaction (PCR-RFLP) assay compared to PapilloCheck® microarray to identify human papilloma virus (HPV) in cervical cells. Three hundred and twenty-five women were analyzed. One sample was used for conventional cytology and another sample was collected using BD SurePath™ kit for HPV tests. Eighty samples (24.6%) were positive for HPV gene by PCR-Multiplex and were then submitted to PCR-RFLP and PapilloCheck® microarray. There was a genotyping agreement in 71.25% (57/80) on at least one HPV type between PCR-RFLP and PapilloCheck® microarray. In 22 samples (27.5%), the results were discordant and those samples were additionally analyzed by DNA sequencing. HPV 16 was the most prevalent HPV type found in both methods, followed by HPVs 53, 68, 18, 39, and 66 using PCR-RFLP analysis, and HPVs 39, 53, 68, 56, 31, and 66 using PapilloCheck® microarray. In the present study, a perfect agreement using Cohen's kappa (κ) was found in HPV 33 and 58 (κ=1), very good for HPV 51, and good for types 16, 18, 53, 59, 66, 68, 70, and 73. PCR-RFLP analysis identified only 25% (20/80) HPV coinfection, and PapilloCheck® microarray found 62.5% (50/80). Our Cohen's kappa results indicate that our in-house HPV genotyping testing (PCR-RFLP analysis) could be applied as a primary HPV test screening, especially in low income countries. If multiple HPV types are found in this primary test, a more descriptive test, such as PapilloCheck® microarray, could be performed.

Cervical cancer in low and middle‑income countries (Review)

“…Thus, immediate action in the public and global health sectors is advised ( 2 , 138 ). Effective precancerous lesion detection using the PapilloCheck ® microarray and the HPV restriction fragment length polymorphism (RFLP) PCR assay was achieved with 62.5 and 25.0% HPV co-infection detection, respectively ( 4 ). The RFLP PCR assay is suggested as a primary HPV test for screening in LMICs, as it is cost-effective, while the PapilloCheck ® microarray assay is considered a more sensitive and descriptive test ( 4 ).…”

“…The most common cause for the occurrence of cervical cancer is a persisting infection with the sexually transmitted human papilloma virus (HPV) (4). HPV is accountable for 90-100% of cervical cancer cases amongst women, especially those <35 years old (5).…”

“…The types of HPV can be classified as either high-risk (HR) or low-risk in terms of their association with precancerous, benign or cancer lesions (6). HR HPV 16 and 18 subtypes are the most prevalent subtypes of HPV, which are responsible for 70% of cervical cancer cases (1,4). In addition, previous studies have identified an association between the HPV 16 and 18 subtypes and malignant tumours of the penis, vulva and anus (1,7).…”

Prevalencia del virus papiloma humano y factores de riesgo asociados en mujeres afiliadas al seguro de salud estatal en Posadas, Misiones (Argentina)

“…En nuestra experiencia, los tipos virales de alto riesgo mostraron mayor prevalencia respecto a los de bajo riesgo y el VPH 16 fue el más frecuente coincidiendo con reportes nacionales e internacionales [17][18][19][20] .…”

“…19 , encontraron 5,3% de VPH 6 en un estudio de mujeres aborígenes del nordeste de Argentina. Golfetto y cols., en un estudio realizado en 325 mujeres que acudieron a un servicio público por un tamizaje cervical en el estado de Santa Catarina, Brasil, encontraron que el genotipo de bajo riesgo prevalente fue el VPH 61 (3,8%) 20 .…”