To investigate the impact of left bundle branch block (LBBB) on sequential single photon emission computed tomography (SPECT)/ CT imaging starting with stress-first SPECT. Consecutive symptomatic low- to intermediate-risk patients without a history of coronary artery disease (CAD) referred for SPECT/CT were included from an observational registry. If stress SPECT was abnormal, additional rest SPECT and, if feasible, coronary CT angiography (CCTA) were acquired. Of the 5,018 patients, 218 (4.3 %) demonstrated LBBB. Patients with LBBB were slightly older than patients without LBBB (65 +/- 12 vs. 61 +/- 11 years, p < 0.001). Stress SPECT was more frequently abnormal in patients with LBBB (82 % vs. 46 %, p < 0.001). After reviewing stress and rest images, SPECT was normal in 43 % of the patients with LBBB, compared to 77 % of the patients without LBBB (p < 0.001). Sixty-four of the 124 patients with LBBB and abnormal stress-rest SPECT underwent CCTA (52 %), which could exclude obstructive CAD in 46 of the patients (72 %). Sequential SPECT/CT imaging starting with stress SPECT is not the optimal imaging protocol in patients with LBBB, as the majority of these patients have potentially false-positive stress SPECT. First-line testing using CCTA may be more appropriate in low- to intermediate-risk patients with LBBB. aEuro cent Stress-first SPECT imaging is attractive if many patients demonstrate normal stress perfusion. aEuro cent The majority of left bundle branch block patients have abnormal stress-first SPECT. aEuro cent Coronary CT excluded obstructive CAD in many LBBB patients with abnormal SPECT. aEuro cent Stress-first SPECT imaging is not the optimal imaging protocol in LBBB patients. aEuro cent In LBBB patients imaging with initial coronary CT may be more appropriate.