This study aimed to validate the single breath count test (SBCT) against volitional measures of respiratory muscle function in healthy subjects and people with neuromuscular disorders (NMD; n = 100 per group). Testing comprised upright and supine SBCT, forced vital capacity (FVC), maximum inspiratory pressure (MIP), and sniff nasal inspiratory pressure (SNIP). Predictability of FVC by SBCT was assessed using logarithmic regression analysis. Receiver operating characteristics curves were used to identify SBCT thresholds for lung restriction (FVC < 80% predicted), inspiratory muscle weakness (MIP < 60 cmH2O), and indication for non-invasive ventilation (NIV) in NMD patients. In both groups, SBCT showed moderate correlation with FVC. In patients, SBCT values were also correlated with MIP and SNIP. Strength of correlations was similar with supine and upright SBCT which accounted for 23.7% of FVC variance in healthy individuals (44.5% in patients). Predictive thresholds of upright SBCT were < 27 for MIP < 60 cmH2O (sensitivity 0.61/specificity 0.86), <39 for NIV indication (0.92/0.46), and <41 for FVC < 80% predicted (0.89/0.62). The SBCT is positively correlated with spirometry. It predicts both lung restriction and NIV indication in NMD patients. The SBCT allows for remote monitoring and may substitute for spirometry/manometry if appropriate devices are unavailable.