The establishment of a multiplex fluorescent polymerase chain reaction coupled with capillary electrophoresis analysis technology enables the simultaneous detection of 16 genotypes of human papillomavirus

Author:

Nie Lingling1ORCID,Qin Haiyang1,Li Sisi2,Yu Zailiang2,Huang Weijin1,Zhang Li1,Zhao Jian3

Affiliation:

1. Division of HIV/AIDS and Sex‐transmitted Virus Vaccines National Institutes for Food and Drug Control Beijing China

2. Suzhou YueMicro Gene Technology Co. Ltd Suzhou China

3. Department of Obstetrics and Gynecology Peking University First Hospital Beijing China

Abstract

AbstractBackgroundThe detection and accurate genotyping of human papillomavirus (HPV) infection is critical for preventing and effectively treating cervical cancer.MethodsA multiplex fluorescent polymerase chain reaction (PCR) coupled with a capillary electrophoresis method was developed for the simultaneous detection of the 16 most prevalent HPV genotypes. Twenty‐five pairs of primers were ultimately selected to ensure that both E and L regions of nine HPV genotypes, as well as the E regions of seven HPV genotypes could be accurately amplified.ResultsThis method enables the simultaneous detection and differentiation of 16 HPV genotypes in a single closed‐tube reaction, accurately distinguishing products with molecular weight differences >1 bp through capillary electrophoresis. This method demonstrated exceptional accuracy, specificity, and repeatability with a detection limit of 10 copies/μL for all 16 HPV genotypes. Furthermore, 152 cervical swab specimens were obtained to compare the disparities between this approach and Cobas 4800 HPV detection method. The concordance rate and κ value were 90.1% and 0.802, respectively, indicating a high level of agreement. The established detection method was successfully applied to cervical swab specimens for determining HPV genotypes across all levels of cervical lesions, HPV52, 56, 16, and 59 were found to be most prevalent with infection rates of 10.8%, 9.1%, 6.5%, and 6.2%, respectively.ConclusionsThis study has successfully established a detection method capable of simultaneously identifying 16 HPV genotypes. This approach can be further applied to HPV vaccine research and surveillance, with the potential for broad applications.

Funder

National Key Research and Development Program of China

Publisher

Wiley

Subject

Microbiology (medical),Biochemistry (medical),Medical Laboratory Technology,Clinical Biochemistry,Public Health, Environmental and Occupational Health,Hematology,Immunology and Allergy

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