Comparison of the performance in detection of HPV infections between the high‐risk HPV genotyping real time PCR and the PCR‐reverse dot blot assays

Abstract

A new multiplex real-time PCR assay, the high-risk HPV genotyping real time PCR assay (HR HPV RT-PCR), has been developed to detect 15 high-risk HPV types with respective viral loads. In this report, a total of 684 cervical specimens from women diagnosed with vaginitis were assessed by the HR HPV RT-PCR and the PCR reaction and reverse dot blot (PCR-RDB) assays, using a PCR-sequencing method as a reference standard. A total coincidence of 97.7% between the HR HPV RT PCR and the PCR-RDB assays was determined with a Kappa value of 0.953. The HR HPV RT PCR assay had sensitivity, specificity, and concordance rates (accuracy) of 99.7%, 99.7%, and 99.7%, respectively, as confirmed by PCR-sequencing, while the PCR-RDB assay had respective rates of 98.8%, 97.1%, and 98.0%. The overall rate of HPV infection, determined by PCR-sequencing, in women diagnosed with vaginitis was 49.85%, including 36.26% of single infection and 13.6% of multiple infections. The most common infections among the 15 high-risk HPV types in women diagnosed with vaginitis were HPV-52, HPV-16, and HPV-58, with a total detection rate of 10.23%, 7.75%, and 5.85%, respectively. We conclude that the HR HPV RT PCR assay exhibits better clinical performance than the PCR-RDB assay, and is an ideal alternative method for HPV genotyping. In addition, the HR HPV RT PCR assay provides HPV DNA viral loads, and could serve as a quantitative marker in the diagnosis and treatment of single and multiple HPV infections. . 2 Meta-analyses have shown that there is a positive association between vaginitis and cervical HPV infection, and that the pooled prevalence of vaginitis can be as high as 32%. [3][4] In addition to single infection, cervical co-infections of multiple HPV genotypes is common. 5 J Med Virol. 2018;90:177-183.wileyonlinelibrary.com/journal/jmv

Cost-effectiveness and accuracy of cervical cancer screening with a high-risk HPV genotyping assay vs a nongenotyping assay in China: an observational cohort study

“…After amplification, HPV genotyping was performed by RDB hybridization on nitrocellulose membrane strips fixed with different HPV type-specific probes. Our study and previous studies [ 17 , 23 ] have demonstrated that the PCR-RDB HR-HPV genotyping assay is an effective HR-HPV genotyping detection method.…”

“…Unit costs were taken from the Fujian medical price database. [17,23] have demonstrated that the PCR-RDB HR-HPV genotyping assay is an effective HR-HPV genotyping detection method.…”

Recent advances in lab-on-a-chip technologies for viral diagnosis

“…Also there is a window period between the viral infection and Table 1 Summary of emerging viruses with abbreviations: molecular (mol), serological (ser), reverse transcription loop-mediated isothermal amplification (RT-LAMP), immunoglobulin type M (IgM), lateral flow assay (LFA), nonstructural protein (NS1), immunoglobulin type G (IgG), loop mediated isothermal amplification (LAMP), digital PCR (dPCR) enzyme-immunoassay (EIA) and IgM antibody capture ELISA (MAC-ELISA), double antibody sandwich ELISA (DAS-ELISA), enzyme linked immunospotting (ELISPOT), data not available (N.A. 1F), dot-blot (Zhang et al, 2018a), or Southern blotting (Cai et al, 2013), but their sensitivity is insufficient. (Hewa et al, 2009;Jung et al, 2015a) antibody production resulting in false negative results using immunoassays, which can be up as long as 35-45 days for first generation HIV testing (Cornett and Kirn, 2013).…”

“…The increase sensitivity acquired in fourth generation of immunoassays led to shortening this window to 10-15 days (Branson and Stekler, 2011). A number of alternative NA techniques have been developed including NA sequence-based amplification (Lanciotti and Kerst, 2001), strand displacement amplification (Shi et al, 2014), or branched DNA probes (Zhang et al, 2018a). Several techniques can directly detect specific viral DNA or RNA via in-situ hybridization (Pfankuche et al, 2018) (Fig.…”

The role of immunocytochemical biomarkers in diagnostics of precancerous pathology of cervix

“…Усі пацієнтки були обстежені на ВПЛ високоонкогенних типів методом полімеразної ланцюгової реакції (ПЛР) з кількісним визначенням [8]. Оцінку експресії імуноцитохімічних біомаркерів р16 і Кі-67 проводили на діагностичному етапі в цитологічному матеріалі [6].…”