Analysis of prostate tissue DNA for the presence of human papillomavirus by polymerase chain reaction, cloning, and automated sequencing

Abstract

We have analysed the DNA from 24 prostate tissue biopsies, spanning a range of Gleason grading from benign to grade 5 and mixed randomly with cervical cancer samples of known human papillomavirus (HPV) status, for the prevalence of HPV DNA, in a double-blind study to ensure complete objectivity. Polymerase chain reactions (PCR) were performed using general E1 open reading frame primers for HPV under low stringency conditions, in addition to reactions containing primers specific for HPV16, E2, and E6 open reading frames under higher, more stringent PCR conditions. The presence of cellular DNA was verified by the use of primers for hypoxanthine guanine phosphoribosyl transferase. DNA bands were not detected in the prostate biopsies using the HPV16-specific primers under high-stringency PCR conditions, however a predominant band in the 400 bp region was observed in 15 of the prostate biopsies using the general primers and the low annealing temperature of 40 degrees C. This fragment was excised and cloned into the pT7 blue vector and the sequence of the insert determined. Although the cloned sequences initiated and terminated with the two authentic PCR primers, they did not contain a significant HPV-related open reading frame. Our results indicate that HPV type 16 and closely related types, as detected by the general primer pair, are unlikely initiators of prostate carcinogenesis within our population.

Association between human papillomavirus and prostate cancer: A meta-analysis

“…These studies were published between 1990 and 2015 and comprised 971 PCa cases in total. The majority of the studies originated from Asia (n=8) (38,40,(43)(44)(45)48,51,56), Europe (n=7) (22,39,42,47,50,53,55) and North America (n=5) (23,49,52,54,57), and the remaining studies originated from South America (n=1) (46) and Oceania (n=2) (6,41 Table I. Characteristics of studies included in the meta-analysis.…”

“…others use frozen sections (6,22,23,43,47,(49)(50)(51)(53)(54)(55). A total of 2,056 cases were included in the 24 studies investigating the association between HPV infection and risk of PCa (971 were in the case group and 1,085 were in the control group).…”

Polymerase chain reaction‐based fluorescent Luminex assay to detect the presence of human papillomavirus types

“…(32,33) The present results are more specifically related to a method for detecting the presence of a HPV type with a plurality of oligonucleotide sets. Each oligonucleotide set of the many available sets was specific to a single gene of the HPV type to be detected.…”

“…We used the HPV PCR-based DNA microarray system as an HPV genotyping method (MyGene Co., Seoul, Korea) for HPV typing. The HPV DNA microarray contains 24 type-specific probes; 16 high-risk types (16,18,31,33,35,39,45, 51, 52, 53, 54, 56, 58, 59, 66 and 68) and seven low-risk types (11, 34, 40, 42, 43, 44 and 70). A slide has eight chambers, and each chamber is used for a test.…”

Viruses in the Mammalian Male Genital Tract and Their Effects on the Reproductive System

“…A secondary adult human prostate epithelial cell culture, on transfection with a plasmid containing the entire type 18 HPV genome, acquired an indefinite, life span in culture but did not undergo malignant conversion (267). Thus, although there are strong arguments in favor of the prostate being an important reservoir for the transmission of HPV (18,101,282,333,350), the involvement of HPV in the pathogenesis of prostate cancer is still a subject of great controversy. For example, two laboratories with extensive experience in HPV research recently tested specimens from two populations with different risks of prostate carcinoma, using three different PCR assays and two serologic assays for HPV.…”