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The reaction was performed using SYBR Green I on a Chromo4 real-time PCR system (Bio-Rad, USA), and relative viral accumulation was calculated based on Ct values

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Complex cases may include: Severe crowding Open bite Crossbite Impacted teeth Missing teeth Rotated teeth Jaw growth concerns Previous orthodontic relapse Cases needing surgical orthodontics In these cases, digital planning may help the orthodontist organize the treatment steps

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