Genotyping of Human Papillomavirus in Liquid Cytology Cervical Specimens by the PGMY Line Blot Assay and the SPF ₁₀ Line Probe Assay

Abstract

A comparison of two PCR-based human papillomavirus (HPV) DNA detection and genotyping systems (PGMY LBA and SPF 10 LiPA) was conducted in two laboratories. Both systems are based on broad-spectrum PCR for the detection of HPV DNA, followed by reverse hybridization with type-specific probes. A total of 400 selected cervical scrape specimens in PreservCyt solution (55% normal cytology, 18% atypical squamous cells of unknown significance, 14.8% low-grade squamous intraepithelial lesions [SIL], and 12.5% high-grade SIL) were tested for the presence of HPV DNA. In this selected group of specimens, the overall agreement between the two methods for the detection of any HPV DNA was high ( ‫؍‬ 0.859). When the 20 common HPV genotypes identified by both methods were considered (HPV types 6, 11, 16, 18, 31, 33, 35, 39, 40, 42, 45, 51, 52, 53, 54, 56, 58, 59, 66, and 68), compatible genotype-specific results were observed in 96.5% of the samples, even when multiple HPV genotypes were present. However, for some specific HPV genotypes, there were significant differences in HPV detection by the two methods. PGMY LBA detected more HPV type 42 (P ‫؍‬ 0.002), HPV type 56 (P ‫؍‬ 0.039), and HPV type 59 (P < 0.001), whereas SPF 10 LiPA detected more HPV type 31 (P < 0.001) and HPV type 52 (P ‫؍‬ 0.031). For the remaining genotypes, including HPV types 16 and 18, the results obtained by the two methods were not significantly different. In general, both genotyping methods are highly suitable for clinical and epidemiological studies.Human papillomavirus (HPV) infection is associated with an increased risk for the development of cervical neoplasia (15,22). Accurate type-specific diagnosis of HPV infections requires sensitive molecular methods, such as PCR. The accurate detection of HPV DNA by PCR is hampered by the existence of a large number of viral genotypes with highly diverse nucleotide sequences (2,5,23,25). PCR-based HPV detection methods have been used for detailed clinical, epidemiological, and natural history studies to elucidate the importance of the different HPV genotypes (7,10,11,21,26). Among the genotypes occurring in the anogenital region, high-risk and low-risk groups have been identified based on their epidemiological association with the development of cervical cancer (4,19,27). Therefore, reliable identification of HPV genotypes, in combination with cytological screening, may be relevant for patient management. In addition, to study the effects of antiviral treatment or type-specific vaccination, accurate HPV genotyping methods are essential for the selection and monitoring of study subjects.Various PCR-based methods have been described for the identification of HPV genotypes. Individual genotypes can be detected by type-specific PCR primer sets (1, 24). However, these require the performance of multiple parallel assays for each sample, and type-specific PCR primers have not been reported for each HPV genotype. Alternatively, general PCR primer sets can be used, permitting simultaneous amplification of a broad rang...

Concurrent Infection with Multiple Human Papillomavirus Types: Pooled Analysis of the IARC HPV Prevalence Surveys

“…Different primers for HPV detection can differ remarkably in their sensitivity to detect multiple infections (43)(44)(45). We showed, however, that the genotyping methods can also substantially affect results and lead to systematic overestimates of certain HPV combinations.…”

Population‐based prevalence, type‐ and age‐specific distribution of HPV in women before introduction of an HPV‐vaccination program in Denmark

“…24 The differences in the prevalence of individual HPV types observed in our study compared to other studies may be because the LiPA test is more sensitive for some HPV types than other HPV detection methods. [25][26][27] In Europe, the 2 HPV types most frequently associated with cervical cancer are HPV 16 and 18 with HPV 16 being the predominant type in squamous cell carcinomas and HPV 18 the predominant type in adenocarcinomas and adenosquamous carci- nomas of the cervix. 7,9 Our study revealed that HPV 16 and/or 18 were present in 6.6% of samples from women with normal cytology, 26.5% of ASCUS/LSIL samples, 19.8% of samples from women with CIN 1 lesions and 63.1% of women with CIN 3 lesions, which is similar to previously published results.…”

Detection and Typing of Human Papillomaviruses Combining Different Methods: Polymerase Chain Reaction, Restriction Fragment Length Polymorphism, Line Probe Assay and Sequencing

“…The question could be raised: are these positive findings by LiPA really HPV 51 or they are results of possible cross-hybridization with other HPV types present in multiple infections. However, HPV 51 was also frequently found in other studies in which other line blot assays for HPV typing [29][30][31][32] were used, which indicate that it can be considered as common HPV type, especially in HSIL specimens.…”

“…This high percentage of multiple infections in HSIL was reported in previous studies. [29][30][31][32] Among the samples analysed by RFLP, LiPA and sequencing, the most frequent low abundant HPV types in single infections in decreasing order were HPV 53, 58, 66, 56 and 52. Similar frequencies of low abundant HPV types in HSIL samples were reported in previous studies.…”