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® 45152D™), HPV-35 (ATCC® 40330™), HPV-43 (ATCC® 40338™) and HPV-56 (ATCC® 40549™) 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 × 101 and 4.68 × 103 copies/μl, respectively. qLAMP assay enabled detection of positive results as early as 23 min at 67 °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 × 101 to 4.68 × 104 copies/μl. 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.
METHODS: We prospectively recruited patients with Adolescent Idiopathic Scoliosis (AIS) scheduled for PSF surgery. The anatomical locations of pain were divided into four: (1) surgical wound pain; (2) shoulder pain; (3) neck pain; and (4) low back pain. The anatomical locations of pain were charted using the visual analogue pain score at intervals of 12, 24, 36, 48 hours; and from day-3 to -14. Patient-controlled analgesia (morphine), use of celecoxib capsules, acetaminophen tablets and oxycodone hydrochloride capsule consumption were recorded.
RESULTS: A total of 40 patients were recruited. Patients complained of surgical wound pain score of 6.2±2.1 after surgery. This subsequently reduced to 4.2±2.0 by day-4, and to 2.4±1.3 by day-7. Shoulder pain scores of symptomatic patients peaked to 4.2±2.7 at 24 hours and 36 hours which then reduced to 1.8±1.1 by day-8. Neck pain scores of symptomatic patients reduced from 4.2±1.9 at 12 hours to 1.8±1.1 by day-4. Low back pain scores of symptomatic patients reduced from 5.3±2.3 at 12 hours to 1.8±1.1 by day- 12.
CONCLUSIONS: Despite the presence of different anatomical locations of pain after surgery, surgical wound was the most significant pain and other anatomical locations of pain were generally mild. Surgical wound pain reduced to a tolerable level by day-4 when patients can then be comfortably discharged. This finding provides useful information for clinicians, patients and their caregivers.