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Published on: March 11, 2014
[Genotyping of human papillomavirus genotyes in cervical scrapes by line probe assay]
J Wang1, J Bakkers, W J Melchers
1Affiliated Hospital of Ningxia Medical College, Yinchuan 750004.
Objective:
To pursue the further development of a practical HPV genotyping system applicable for mass screening.
Methods:
Cervical scrapes from 155 women were tested for the presence of HPV using a general-primer based PCR. A reverse hybridization assayor, the line probe assay (LiPA) was used for genotyping 16 different HPV types(HPV 6, 11, 16, 18, 31, 33, 35, 40, 42, 43, 44, 45, 51, 52, 56, 58) simultaneously.
Results:
93(60%) out of 155 specimens tested were positive for HPV DNA by PCR. The HPV positivity amongst the group of women with no cervical abnormality was 43%, while that in women with cervical dysplasia and cervical cancer was 81.2% PCR products from 89 HPV positive cases were genotyped by LiPA, of which 49 cases were single HPV infected concerning 11 different HPV types and the commonest type was HPV-16, occurring in 25(51%) of all single HPV infected cases. The other 40 specimens contained multiple HPV types comprising 16 different HPV types. Although double infections(n = 25) prevailed, triple(n = 9), quadruple(n = 3) and quintuple (n = 3) infections were also found. PCR products of four HPV positive cases failed to be classified by LiPA. Sequencing analysis revealed that these 4 cases were HPV-66 which was not included in LiPA probes. Follow-up data of 12 cases confirmed the HPV genotyping results.
Conclusion:
The LiPA for HPV genotyping is easy to perform. It allows accurate and rapid identification of 16 different HPV types in cervical scrapes and will facilitate HPV detection as well as genotyping in cervical cancer screening programs.