Please use this identifier to cite or link to this item:
|Title:||Sequence variants of human papillomavirus type 16 in clinical samples permit verification and extension of epidemiological studies and construction of a phylogenetic tree||Authors:||Ho, L.
|Issue Date:||1991||Citation:||Ho, L.,Chan, S.-Y.,Chow, V.,Chong, T.,Tay, S.-K.,Villa, L.L.,Bernard, H.-U. (1991). Sequence variants of human papillomavirus type 16 in clinical samples permit verification and extension of epidemiological studies and construction of a phylogenetic tree. Journal of Clinical Microbiology 29 (9) : 1765-1772. ScholarBank@NUS Repository.||Abstract:||Genomic variability between different viral isolates provides a powerful epidemiological tool for verifying ultrasensitive diagnostic procedures, understanding infectious pathways in individuals and human populations, and studying viral evolution. The potential of this approach has not yet been exploited for the diagnosis of human papillomaviruses (HPVs) like HPV type 16 (HPV-16), which are involved in genital cancer. Toward this end, we amplified by polymerase chain reaction, cloned, and sequenced a 364-bp noncoding segment of the HPV-16 genome from cell lines, cervical biopsy specimens, and cervical smears. The HPV-16 genomes in the cell lines SiHa and CaSki showed an identical point mutation, and in the SiHa cell line it had an additional 38-bp deletion. Only 4 of 22 cervical lesions biopsied from patients at several hospitals in Singapore contained HPV-16 DNA with the prototype sequence, while the DNAs of the other 18 cervical lesions differed by 1 to 10 mutations. This excludes contaminations with cloned HPV-16 DNA as the source of this DNA. To test whether this diversity was a geographic idiosyncrasy, we analyzed 25 cervical biopsy specimens from Brazil. Eight of these contained the prototype sequence, while 17 were mutated. Altogether, 11 genomic variants were found in the Singaporean samples and 12 genomic variants were found in the Brazilian samples, and only 5 of these occurred identically in both cohorts. All variants could be connected to form a phylogenetic tree, with some branches being specific for each cohort. This suggests that the variants did not originate over a short period in the individual patient but, rather, evolved consecutively while spreading throughout humankind. Repeated amplification with the low-error Vent polymerase suggested that at least five patients each had two variants in the same biopsy specimen, while another patient exhibited different variants in two separate biopsy specimens. This documents the possibility of multiple infections with variants of the same papillomavirus type. We do not believe that the observed variants have greater carcinogenicity, since similar variants were found in cervical smears from asymptomatic women. In chloramphenicol acetyltransferase assays, enhancer segments of some selected samples that were analyzed did not show altered transcriptional activity. This was expected since most of the mutations did not coincide with known cis-responsive elements. The observed sequence variability can be used as a natural tag of the viral genome to unambiguously assess a variety of epidemiological questions.||Source Title:||Journal of Clinical Microbiology||URI:||http://scholarbank.nus.edu.sg/handle/10635/112069||ISSN:||00951137|
|Appears in Collections:||Staff Publications|
Show full item record
Files in This Item:
There are no files associated with this item.
checked on Jul 12, 2021
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.