Abstract:
Objective To established a real-time polymerase chain reaction assay with high specificity for the detections of genotypes of human papillomavirus (HPV).
Methods Specific primers and minor groove binder (MGB) probes were designed for 23 HPV genotypes. The primer-probe sequences and reaction system were optimized. The second-generation national reference for HPV genotyping, published by the China National Institute for Food and Drug Control, was used to evaluate the effectiveness, sensitivity, precision, and cross-specificity of the assay. The assay was then applied in the cervical cell screening of patients with known subtypes.
Results All the 23 primers and binders were tested for validity, sensitivity, cross-specificity and precision. In sensitivity test, the minimum detection limit could reach 100 copies/µL, and the coefficient of variation was ≤3.18%. Of the 114 clinical samples, 100 were positive and 14 were negative, all were consistent with known results.
Conclusion The genotype detection system established in this study can specifically detect 23 HPV species and identify their genotypes, which can be used as a potential ideal detection method to provide effective reference for the clinical diagnosis of HPV infection.
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