Plasma HPV DNA is detectable in oral leukoplakia patients

Abstract

Based on the present study, there was no difference in the rate of HPV detection in patients with or without oral leukoplakia. However all sources tested in this study were considered suitable for HPV detection, especially plasma samples, which showed be an important non-invasive source of HPV detection in leukoplakia patients.

Prevalence of human papillomavirus type 16 in persistent oral lesions arising in patients with tobacco, areca nut, and alcohol habits

“…In the present work, HPV DNA was found in normal mucosa (C_1N), implicating the existence of HPV species in the oral cavity of healthy nonaddict individual. Conjointly, 39 freshly collected oral biopsies were checked for HPV viral DNA using the conventional PCR method involving degenerate (MY09/MY11) and nested primers (GP05+/GP06+) targeting HPV L1 gene, which is capable of amplifying and identifying multiple HPV types; the nucleotide sequence of the L1 gene is highly conserved across HPVs and forms the basis for HPV genotyping . HPV viral DNA was detected in the POLs of all clinical cases (n = 39) comprising OSCC (n = 24) and precancer non‐OSCC (n = 15) lesions.…”

“…The presence of HPV viral DNA was assessed by conventional PCR using degenerate primers MY09 and MY11 (Table ) targeting the L1 gene (HPV species) in reaction mixture composing nuclease‐free water, Taq buffer (1X), MgCl 2 (1.5 mmol/L), dNTPs (0.2 mmol/L), primers (0.4 µmol/L), DNA (60 ng), and Taq polymerase (1 U); the thermocycling steps were as follows: (a) initial denaturation: 95°C for 5 minutes; (b) 40 cycles of denaturation (95°C for 1 minute), annealing (56°C for 1 minute) and extension (72°C for 1 minute); and (c) final extension: 72°C for 10 minutes. Subsequently, a nested PCR assay was carried out using GP05+ and GP06+ specific primers (Table ), targeting the L1 amplicons amplified using MY09/MY11, for confirming the presence of HPV DNA using the reaction mixes comprising nuclease‐free water, Taq buffer (1X), MgCl 2 (1.5 mmol/L), dNTPs (0.2 mmol/L), primers (0.4 µmol/L), Taq polymerase (1 U), and DNA template (5 µL amplification product of MY09/MY11); the thermocycling steps were as follows: (a) initial denaturation: 95°C for 5 minutes; (b) 40 cycles of denaturation (95°C for 1 minute), annealing (45°C for 1 minute) and extension (72°C for 1 minute); and (c) final extension: 72°C for 10 minutes. The HPV16 E6 gene‐specific primers HPV16‐E6F and HPV16‐E6R (Table ) were used for detecting the presence of HPV16 DNA using reaction mixes comprising: nuclease‐free water, Taq buffer (1X), MgCl 2 (1.5 mmol/L), dNTPs (0.2 mmol/L), primers (0.4 µmol/L), DNA (60 ng), and Taq polymerase (1U); the thermocycling steps were as follows: (a) initial denaturation: 95°C for 5 minutes; (b) 45 cycles of denaturation (95°C for 1 minute), annealing (52°C for 1 minute) and extension (72°C for 1 minute); and (c) final extension: 72°C for 10 minutes.…”

Assessment of cellular and serum proteome from tongue squamous cell carcinoma patient lacking addictive proclivities for tobacco, betel nut, and alcohol: Case study

“…The PCR compatibility of genomic DNA was checked by PCR with β‐globin specific primers (Table ) . The presence of HPV DNA, in the isolated genomic DNA samples, was checked by PCR involving primers MY09/MY11 and GP05+/GP06+ (Table ) . The presence of HPV16 was assessed by using specific primers E6HPV16‐F/E6HPV16‐R (Table ) .…”

“…[59][60][61] Subsequently, the presence of HPV was confirmed by performing nested PCR with GP05 +/GP06+ primers; each reaction constituted of 5 μL of MY09/MY11 amplification product, 1× Taq buffer, 1 U Taq polymerase, 1.5 mM MgCl 2 , 0.4 mM dNTPs, 0.4 μM of each primers (Table 1); thermal conditions were (i) 95°C for 5 min (initial denaturation), (ii) 35 cycles at 95°C for 30 s, 45°C for 30 s, and 72°C for 30 s, and (iii) final extension (72°C for 7 min). 61,62 The presence of HPV16 was assessed by using specific primers targeting the long control region (LCR); each reaction constituted of 40 ng DNA, 1× Taq buffer, 1 U Taq polymerase, 1.5 mM MgCl 2 , 0.4 mM dNTPs, 0.4 μM of each primer (Table 1); thermal conditions were (i) 95°C for 5 min (initial denaturation), (ii) 45 cycles at 95°C for 30 s, 56°C for 30 s, and 72°C for 30 s, and (iii) final extension (72°C for 7 min). 59 The integration status of HPV16 DNA was assessed by PCR with E2 and E6 specific primers.…”

“…58,59 The presence of HPV DNA, in the isolated genomic DNA samples, was checked by PCR involving primers MY09/MY11 and GP05+/GP06+ (Table 1). [59][60][61][62] The presence of HPV16 was assessed by using specific primers E6HPV16-F/E6HPV16-R (Table 1). 63 All reactions (25 μL volume) were performed in triplicates on the Thermo Scientific Arktik thermal cycler (Thermo Fisher Scientific).…”