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Background and Objective: Previous studies demonstrated that the prevalence of lumbar disc herniation (LDH) is relatively high. This investigation aimed to evaluate the size of lumbar multifidus (LM) muscle in patients with different degrees of LDH compared to healthy group, during rest and contraction, using ultrasonography. Materials and Methods: In this non-experimental, analytic, and case control study, ultrasound imaging was used to assess cross-sectional area (CSA) and thickness of the LM muscle in 15 healthy subjects and 60 patients with different stages of LDH (bulging group = 15, protrusion group = 15, extrusion group = 15, sequestration group = 15). Measurements were taken bilaterally at the L4-L5 level, during rest and contraction and results were compared between groups. Results: There was a significant difference between healthy subjects and the extrusion and sequestration groups during rest and contraction for LM muscle CSA and thickness (p = 0.001), as LM muscle CSA and thickness were significantly smaller in extrusion and sequestration patient groups compared to healthy subjects. LM atrophy was greater in patients with extrusion and sequestration groups than in patients with bulging and protrusion, both at rest and during contraction. Significant correlations were also observed between functional disability and intensity of pain with LM CSA and thickness measurements. Conclusions: Patients with extrusion and sequestration LDH had smaller LM muscle at rest and during contraction compared to healthy subjects. Larger LDH lesions were associated with decreased LM muscle size. Patient with more pain, disability, and extrusion and sequestration LDH had greater LM size changes. LM muscle size was not correlated with symptom duration. Further investigation with greater sample size is warranted.