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1.
Int J Cancer ; 136(11): 2546-55, 2015 Jun 01.
Article in English | MEDLINE | ID: mdl-25388227

ABSTRACT

Non-melanoma skin cancers commonly contain Human Papillomavirus (HPV), but the types found have varied depending on the polymerase chain reaction (PCR) primer systems used. Whole genome amplified DNA (not amplified by any specific PCR primers) from 91 skin lesions [41 squamous cell skin carcinomas (SCCs), 8 keratoacanthomas, 22 actinic keratoses, 3 basal cell carcinomas and 17 SCCs in situ] were sequenced. All samples were sequenced both at 160 Mb and 1.8 Gb sequencing depth per sample. The sequences from 10 different HPVs in 47/91 specimens were found. Sequences represented four established HPV types (HPV types 16, 22, 120, 124), two previously known putative types (present in GenBank) and four previously unknown HPV sequences (new putative types). The most commonly detected virus was cloned, sequenced and designated as HPV197. Type-specific real-time PCR detected HPV197 in 34/91 specimens. For comparison, a pool of the same samples after general primer PCR amplification was also sequenced. This revealed 40 different HPVs, but only two HPV types were detected both with sequencing without prior PCR and with sequencing PCR amplicons, suggesting that sequencing without prior PCR gives a more unbiased representation of the HPVs present. In summary, it was found that HPV can be sequenced from most skin disease specimens and HPV197 appeared to be the most commonly present virus.


Subject(s)
Papillomaviridae/classification , Papillomaviridae/isolation & purification , Skin Neoplasms/virology , Carcinoma, Basal Cell/genetics , Carcinoma, Basal Cell/virology , Carcinoma, Squamous Cell/genetics , Carcinoma, Squamous Cell/virology , Cloning, Molecular , DNA, Viral/genetics , Humans , Keratoacanthoma/genetics , Keratoacanthoma/virology , Keratosis, Actinic/genetics , Keratosis, Actinic/virology , Molecular Sequence Data , Papillomaviridae/genetics , Sequence Analysis, DNA , Skin Neoplasms/genetics , Skin Neoplasms/pathology
2.
J Med Virol ; 86(12): 2042-8, 2014 Dec.
Article in English | MEDLINE | ID: mdl-24532541

ABSTRACT

To investigate which microorganisms may be present in expressed prostate secretions (EPS) metagenomic sequencing (MGS) was applied to prostate secretion samples from five men with prostatitis and five matched control men as well as to combined expressed prostate secretion and urine from six patients with prostate cancer and six matched control men. The prostate secretion samples contained a variety of bacterial sequences, mostly belonging to the Proteobacteria phylum. The combined prostate secretion and urine samples were dominated by abundant presence of the JC polyomavirus, representing >20% of all detected metagenomic sequence reads. There were also other viruses detected, for example, human papillomavirus type 81. All combined prostate secretion and urine samples were also positive for Proteobacteria. In summary, MGS of expressed prostate secretion is informative for detecting a variety of bacteria and viruses, suggesting that a more large-scale use of MGS of prostate secretions may be useful in medical and epidemiological studies of prostate infections.


Subject(s)
Bodily Secretions/microbiology , Bodily Secretions/virology , Metagenomics , Prostatic Neoplasms/microbiology , Prostatic Neoplasms/virology , Prostatitis/microbiology , Prostatitis/virology , Adult , Bacteria/classification , Bacteria/isolation & purification , Humans , Male , Pilot Projects , Sequence Analysis, DNA , Urine/microbiology , Urine/virology , Viruses/classification , Viruses/isolation & purification
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