Block Details : Total number of blocks received in RGCB –1004

1. Role of Human PapillomaVirus infection and other co factors in the aetiology of Head and Neck Cancer in Europe and India

2. Block Details : • Total number of blocks received in RGCB –1004 • Different sites from which we received the blocks are: • Ambilikkai -148 • Bangalore- 344 • Guwahati-346 • Barshi-156 • Trivandrum -10

3. Serum transferred to RGCB -540 (RCC, Trivandrum) • Total number of blocks sectioned -69 • Blocks returned back to Bangalore after sectioning: 25 • Blocks take n to IARC: Prospective-68(Ambilikkai), Retrospective-80(Ambilikkai) • Guwahati -40 • Total : 188 • Total number of shavings taken to IARC: 116 tubes and 13 boxes

4. Oral Saline washouts : • Pune: 320 • Ambilikkai:216 • Barshi: 341 • DNA isolated: 877

5. Head and Neck Cancer is the 6th common malignancy reported worldwide with approximately 6,50000newcases and 3,50000 deaths are reported annually • Cancers of the head and neck include cancers of the oral cavity, larynx, pharynx, salivary glands and nasal passage • The prevalence of HPV in oral cavity, larynx and hypopharynxappears to be considerably lower • Oral cancer subtype of HNC(85%) • Several types of oral cancer ,around 90% squamous cell carcinoma originating in the tissue that line mouth and lips

6. Causative factors of oral cancer: • Alcohol abuse, radiation exposure, tobacco use, poor nutrition and poor hygiene • HPV is considered to be one of the factor • Symptoms include lump or sore that does not heal, difficulty in swallowing, change of hoarseness in the voice

7. Human papilloma virus: • Papillomaviridae family, double stranded DNA virus, infects cutaneous/mucosal surfaces • Infects basal layer of the skin • Genome: 8000basepairs, divided into 3 main domains a noncoding URR, an early region with ORF,and late region encoding capsid proteins • E6 and E7 binds to the tumor supressor gene pRb and p53

8. Objective of the study: • To address the crucial still unanswered questions on HNC aetiology with particular focus on the role of HPV infection • To determine whether HPV interact with additional HNC risk factors in Europe and Indian population • To know prevalence of DNA/RNApositivity in oral specimens and also to explore novel screening stratergies that could be broadly used in clinical routine

9. Materials and Methodology: • Total study size 2000 cases and 2000controls • Cases include cancers of the oral cavity , oropharynx, hypopharynx, and larynx • Samples include serum, oral saline wash and formalin fixed paraffin embedded tissue

10. Methodology: • Serology • Genotyping • Immunohistochemistry • Luminexxmap technology

11. Genotyping: • The process of determining difference in the genetic makeup of an individual by examining the individual DNA sequence using biological assays and comparing to another individual sequence for reference • Luminexxmap technology for detecting 19 high risk HPV types (16,18,26,31,33,35,39,45,51,52,53,56,58,59,66,68,70,73,82)and 2 low risk hpv types(6,11) • The assay includes Beta globin gene to evaluate the presence and quality of extracted DNA

12. Outline • Luminex® xMAP Technology

13. DETECTION OF THE BEADS USING LASER