Head‐to‐head comparison of genotyping of human papillomavirus by real‐time multiplex PCR assay using type‐specific primers and SPF10‐PCR‐based line probe assay

Author:

Yin Jian12ORCID,Peng Siying23,Zhang Changning24,Li Xinyue24,Hu Fangfang23,Chen Wen2ORCID,Qiao Youlin1

Affiliation:

1. School of Population Medicine and Public Health Chinese Academy of Medical Sciences and Peking Union Medical College Beijing China

2. Department of Cancer Epidemiology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital Chinese Academy of Medical Sciences and Peking Union Medical College Beijing China

3. The State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, Collaborative Innovation Center of Biologic Products, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health Xiamen University Xiamen Fujian China

4. College of Life Sciences Hebei University Baoding China

Abstract

AbstractThe SPF10‐polymerase chain reaction (PCR)‐based line probe assay (LiPA‐25) with high analytical sensitivity and specificity for human papillomavirus (HPV) genotyping in clinical samples has been widely used in vaccine and epidemiologic studies. A real‐time multiplex PCR assay using type‐specific primers (Hybribio‐23) with low workload and cost has been developed recently. The study aimed to compare the performance of LiPA‐25 and Hybribio‐23 in selected 1731 cervical swab and 117 tissue samples, with a focus on 20 common HPV types (14 high‐risk: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68/73; 6 low‐risk: 6, 11, 42, 43, 44, and 53). The level of agreement of two assays was determined using Cohen's Kappa (κ) statistics. A total of 1296 (74.9%) swab samples were identified as HPV‐positive by Hybribio‐23 or LiPA‐25, of which 814 (62.8%) samples exhibited concordant, 358 (27.6%) showed additional or fewer types (compatible), and 124 (9.6%) were discordant. In addition, the two assays showed a perfect agreement for 20 HPV‐combined detection (κ = 0.838) and 17 individual HPV types (all κ > 0.800), a good agreement for HPV31 (κ = 0.792) and 43 (κ = 0.696), and a moderate agreement for HPV42 (κ = 0.504). Hybribio‐23 was significantly more sensitive for HPV58, 59, 68/73, 42, 43, and 44, and less sensitive for HPV35 and 66 than LiPA‐25 (McNemar's test: all p < 0.05). For 117 HPV‐positive tissue specimens, the identification of genotypes was 85.2% identical, 12.2% compatible, and only 2.6% discordant. The agreement for HPV31 (κ = 0.786), 68/73 (κ = 0.742), and HPV53 (κ = 0.742) was good, while for other types (all κ > 0.853) and 20 HPV‐combined detection (κ = 0.936) was perfect (all p > 0.05). In conclusion, Hybribio‐23 and LiPA‐25 are comparable. Hybribio‐23 could be used for the detection and genotyping of HPV in cervical samples for epidemiological and vaccine studies worldwide.

Publisher

Wiley

Subject

Infectious Diseases,Virology

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