Comparison of Polymerase Chain Reaction and Immunohistochemistry Assays for Analysing Human Papillomavirus Infection in Oral Squamous Cell Carcinoma

Abstract

Introduction: Polymerase Chain Reaction (PCR) and Immunohistochemistry (IHC) are two well-known techniques used for the diagnoses of genetic diseases, tumours and different pathogens. PCR basically amplify regions of DNA within a single molecule which may have etiologic significance, it is a method for in vitro amplification of specific DNA or RNA sequences, whereas IHC is used to verify tissue constituents (the antigens) with the utilization of specific antibodies that can be visualized through staining. Aim:To compare and analyse PCR and IHC for their sensitivity to detect Human Papillomavirus (HPV) in Oral Squamous Cell Carcinoma (OSCC). Materials and Methods:This study was based on samples retrospectively collected from 47 patients with primary OSCC who were diagnosed and treated at The Aga Khan University Hospital, Karachi, Pakistan, during the period of January 2010 to December 2013. Inclusion criteria were complete clinicopathologic data, adequate clinical follow up and availability of sufficient paraffin-embedded tumour material. HPV general and type specific 16 and 18 were investigated by means of PCR. HPV immunoreactivity was further investigated by means of IHC.Results: Among the 47 evaluated patients, 32 (68.1%) were male and 15 (31.9%) were female, PCR detected the presence of HPV in 32 (68.1%) patients while IHC showed no positive test results. p53 was positive in 32 (68.1%) patients and negative in 15 (31.9%). HPV type 16 being most prevalent showing positivity in 27 (57.4%) patients whereas, type 18 was positive in only 1 (2.1%) patient. Conclusion:We concluded that PCR is more sensitive and reliable when diagnosing and detecting HPV for OSCC rather than IHC as results from IHC were all negative and insignificant, hence PCR should be the first initial diagnostic test for detecting HPV due to its better sensitivity and successful detection of HPV.www.jcdr.net Muhammad Sohail Awan et al., Comparing PCR and IHC in analyzing HPV in OSCC

Human papillomavirus prevalence in a Mid‐European oral squamous cell cancer population: A cohort study

“…This was particularly evident for subjects with high‐risk HPV infection who were nearly 10 years younger. Previous studies already correlated age of OSCC patients at first diagnosis and oral HPV status, confirming that HPV‐positive cases are younger than HPV‐negative ones (Awan, Irfan, Zahid, Mirza, & Ali, ; Gonzalez‐Ramirez et al., ). However, the difference between both groups has not been quantified as performed in the present study.…”

Clinical efficacy of an antibody-based detection system for human papilloma virus infection in oral squamous cell carcinoma

“…The differential clinical outcome with potential therapy alterations led to an update of the TNM classification in the UICC 8th edition of the Union for International Cancer Control/American Joint Committee on Cancer for oropharyngeal carcinoma, where HPV status was implemented [12]. Today, several methods for HPV detection are frequently used in the clinical setting, with detection of HPV E6/E7 RNA expression, indicating active viral oncogene transcription in tumor cells, as the most accurate testing method [13][14][15]. However, because RNA isolation requires additional sample preparation steps and a larger number of tumor cells, the most widely used assay is the cyclin-dependent kinase inhibitor 2A oncoprotein (p16) expression by immunohistochemistry (IHC) staining, which displays a surrogate marker of oncogenic HPV infection, mainly for OPSCC [16].…”

Human papillomavirus detection in oropharyngeal squamous cell carcinoma using p16 immunohistochemistry

“…polymerase chain reaction (PCR) was a commonly used test, and it has been proved to be more sensitive in detection of HPV in OPSCC, than IHC and in situ hybridization (ISH). [81920]…”

“…[67142025] Studies comparing the results of HPV + in OPSCC revealed using three methods (IHC, ISH, and PCR) explained that cases which were HPV positive by PCR\ISH were also p16 positive by IHC. [141920] The detection of p16 by IHC showed a very high level of sensitivity but less level of specificity. [62025] p16 or HPV E6\E7 mRNA expression was thought to be the parameter to describe the activity of viral oncogenes but a finding that exactly explained that the p16\HPV DNA (+) events were the results of HPV inactive infection.…”