Real-time loop-mediated isothermal amplification assay for rapid detection of human papillomavirus 16 in oral squamous cell carcinoma

Hamzan, Nurul Izzati and Ab Rahman, Nurhayu and Suraiya, Siti and Mohamad, Irfan and Kalarakkal, Thomas George and Mohamad, Suharni (2021) Real-time loop-mediated isothermal amplification assay for rapid detection of human papillomavirus 16 in oral squamous cell carcinoma. Archives of Oral Biology, 124. ISSN 0003-9969, DOI

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Objective: The present study established a real-time loop-mediated isothermal amplification (qLAMP) for rapid detection of human papillomavirus subtype 16 (HPV-16) in oral squamous cell carcinoma (OSCC). Methods: The qLAMP assay was optimized targeting the HPV-16 E7 gene. The analytical sensitivity and specificity of the assay were determined using HPV-18 (ATCC (R) 45152D T), HPV-35 (ATCC (R) 40330 T), HPV-43 (ATCC (R) 40338 T) and HPV-56 (ATCC (R) 40549 T) viral strains and oral bacteria. HPV-16 standard curve was constructed for determination of HPV-16 viral load. The diagnostic performance of the assay was evaluated from 63 OSCC patients comprising 63 tissue, 13 saliva and 49 blood samples, in comparison with p16 immunohistochemistry (IHC), in-house PCR and nested PCR assays. Results: The detection limit of developed LAMP and PCR assays was 4.68 x 10(1) and 4.68 x 10(3) copies/mu l, respectively. qLAMP assay enabled detection of positive results as early as 23 min at 67 degrees C. This assay can detect HPV-16 positivity in 23 % (3/13) saliva and 4.8 % (3/63) tissue samples with the viral load ranging from 4.68 x 101 to 4.68 x 104 copies/mu l. HPV-16 positivity was not detected in all the blood samples. The sensitivity and specificity of qLAMP were 100 % in comparison with that of p16 IHC and nested PCR. Conclusion: This study reports for the first time on the use of qLAMP assay for detection of HPV-16 in OSCC in both tissue and saliva as the sample matrix which holds promise in improving the diagnostic application owing to its rapidity, simplicity, high sensitivity and specificity.

Item Type: Article
Funders: Ministry of Energy, Science, Technology, Environment and Climate Change (MESTECC), Malaysia [02-0105-SF0710], Ministry of Education, Malaysia
Uncontrolled Keywords: Human papillomavirus; Oral squamous cell carcinoma; Real-time loop-mediated isothermal amplification
Subjects: R Medicine > R Medicine (General)
R Medicine > RK Dentistry
Divisions: Faculty of Dentistry > Dept of Oral & Maxillofacial Surgery
Depositing User: Ms Zaharah Ramly
Date Deposited: 04 Aug 2022 08:06
Last Modified: 04 Aug 2022 08:06

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