The last cycle was extended for 10 min at 72☌. Ten minutes of denaturation at 94☌ for the first cycle, followed by 1 min each of denaturation at 94☌, annealing at 48☌, and extension at 72☌ for 33 cycles was done. Reaction was performed in a DNA thermal cycler (Eppendorf, Germany) as per the understated protocol. The plasmid DNA containing HPV genome types 6 and 11 was used as a positive control in the reaction. Three microliters of template DNA was added for each reaction. The reaction was carried out in a volume of 20 µL containing 2 µL 10X Taq buffer, 10 pmol of each of the sense and antisense primers 250 µmol dNTP mix, 1.5 units of Taq polymerase (Invitrogen, USA), and distilled water. The sequence of the forward primer was 5´- TATGGCTATTCTGAAGTGGAA-3´ and that of the reverse primer was 5´-GATATACCTGTTCTAAACCA-3´. ![]() The prepared DNA was quantitated spectrophotometrically, and PCR for b-actin gene was done for each sample as control reaction to check the quality of material.Į1-PCR analysisAll the samples were subjected to PCR using primers specific for E1 open-reading frame of the HPV genome. After phenol-chloroform extraction, the DNA was precipitated with chilled isopropanol and resuspended in TE buffer. In a few cases, biopsies from two different skin tags of the same patient were analyzed.ĭNA extractionBriefly, the tissue specimen was teased and suspended in 500 µL of lysis buffer containing 1% sodium dodecyl sulphate (SDS) and 0.01% proteinase K in Tris-EDTA (TE) buffer (pH 8.0) and incubated at 55☌ overnight. Ten biopsy specimens from the normal skin surrounding skin tags were also analyzed. After obtaining informed consent, biopsy specimens from skin tags were obtained from neck, dorsum of hand and axilla from 37 patients. Those subjects who presented to the dermatology outpatient clinic with skin tags on multiple sites and were otherwise healthy and willing to participate in the study were enrolled. We undertook the present study with the aim of establishing the presence of low-risk human papillomavirus types 6 and 11 using polymerase chain reaction (PCR) systems in a benign cutaneous lesion like skin tag in a tertiary care hospital setting in India. Certain HPV types are found to be associated with the pathogenesis of benign lesions like papillomas of larynx, conjunctiva respiratory papillomatosis etc. Fibroepithelial polyps, or acrochordons, often develop in areas of skin friction. There is a loose, edematous fibrovascular core with mild chronic inflammation. ![]() Histologically, skin tag is a polypoid lesion with overlying mildly acanthotic epidermis. Skin tag, or soft fibroma, is a common benign condition, which consists of a bit of skin which projects from the surrounding skin. The HPV genome usually remains episomal but in transformed cells, the viral DNA is frequently integrated into the host DNA. Types with high oncogenic potential (which always express the early proteins E6 and E7) are able to transform keratinocytes on their own. The numerous HPV types differ in their biological properties and oncogenic potential. In healthy population, most of these HPV types appear to establish a latent infection of the skin, mostly as normal flora residing in hair follicles. Over 100 HPV types have been identified till date. Human papillomavirus (HPV) is known to be the most ubiquitous of the human viruses. The identification of HPV 6/11 in these lesions, which are benign proliferations of the skin, further expands the spectrum of HPV-linked lesions. Conclusion: This result corroborates the hypothesis that HPV plays a part in the etiology of benign lesions like cutaneous soft fibromas. Results: The results revealed the presence of HPV DNA 6/11 in 48.6% of the skin tags examined by PCR-RFLP. Highly sensitive and comprehensive polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) assays were done for the detection of low-risk HPV types 6 and 11. Methods: A total of 37 cases of skin tags from various sites were analyzed. Aims: The present study was designed to detect the existence of low-risk HPV types 6 and 11 in cutaneous soft fibromas (skin tag) in north Indians. In an isolated report available, HPVs have been implicated in the causation of skin tags too. ![]() Abstract Background: Low-risk human papillomavirus (HPV) infections are related to the genesis of various benign lesions.
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