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 https://doi.org/10.1016/j.archoralbio.2021.105051.
Full text not available from this repository.Abstract
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 |
| URI: | http://eprints.um.edu.my/id/eprint/28423 |
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