The objective of this study was to evaluate the California Mastitis Test (CMT) and a portable electrical conductivity meter for diagnosing precalving intramammary infection (IMI) in Holstein heifers. A total of 428 dairy heifers from 23 dairy herds were enrolled between 6 and 12 days before the expected calving date from June 2002 to June 2003. Mammary secretions were tested by both diagnostic methods and by bacterial culture for evidence of IMI. California Mastitis Test was considered negative if the score was negative, trace or 1 and was considered positive otherwise. Two cut-off points were evaluated for milk electrical conductivity (>5 and >6.5 mS/cm). From this study, an overall proportion of 69% of heifers had precalving IMI and the overall heifer prevalence of major pathogen IMI was 16.8%. At the quarter level, sensitivity and specificity of CMT (68.9% and 68.4%, respectively) and milk conductivity >5 mS/cm (41.0% and 65.2%, respectively) or >6.5 mS/cm (25.2% and 83.3%, respectively) to identify all IMI were low. However, the heifer level sensitivity and specificity of CMT for major pathogens were 91.0% (81.5-96.6) and 27.5% (22.8-32.6), respectively. Using a cut-off point of 5 mS/cm, the heifer level sensitivity and specificity for major pathogens was 68.7% (56.2-79.4) and 44.1% (38.7-49.6), respectively. A conductivity cut-off value of 6.5 mS/cm decreased the sensitivity and increased the specificity to 53.7% (41.1-66.0) and 59.5% (54.0-64.8), respectively. California Mastitis Test and milk electrical conductivity are not good predictors of major pathogen IMI in heifers during the last 2 weeks before calving. However, the negative predictive values at quarter or heifer level were high and the heifer false negative rate was 6-14% using CMT or conductivity, respectively. Therefore, these measures could be useful for screening out heifers or quarters that are unlikely to have a major pathogen IMI.