For a national survey in which wide geographic region or an entire country is targeted, multi-stage sampling approach is widely used to overcome the problem of simple random sampling, to consider both herd- and animallevel factors associated with disease occurrence, and to adjust clustering effect of disease in the population in the calculation of sample size. The aim of this study was to establish sample size for estimating bovine tuberculosis (TB) in Korea using stratified two-stage sampling design. The sample size was determined by taking into account the possible clustering of TB-infected animals on individual herds to increase the reliability of survey results. In this study, the country was stratified into nine provinces (administrative unit) and herd, the primary sampling unit, was considered as a cluster. For all analyses, design effect of 2, between-cluster prevalence of 50% to yield maximum sample size, and mean herd size of 65 were assumed due to lack of information available. Using a two-stage sampling scheme, the number of cattle sampled per herd was 65 cattle, regardless of confidence level, prevalence, and mean herd size examined. Number of clusters to be sampled at a 95% level of confidence was estimated to be 296, 74, 33, 19, 12, and 9 for desired precision of 0.01, 0.02, 0.03, 0.04, 0.05, and 0.06, respectively. Therefore, the total sample size with a 95% confidence level was 172,872, 43,218, 19,224, 10,818, 6,930, and 4,806 for desired precision ranging from 0.01 to 0.06. The sample size was increased with desired precision and design effect. In a situation where the number of cattle sampled per herd is fixed ranging from 5 to 40 with a 5-head interval, total sample size with a 95% confidence level was estimated to be 6,480, 10,080, 13,770, 17,280, 20.925, 24,570, 28,350, and 31,680, respectively. The percent increase in total sample size resulting from the use of intra-cluster correlation coefficient of 0.3 was 22.2, 32.1, 36.3, 39.6, 41.9, 42.9, 42,2, and 44.3%, respectively in comparison to the use of coefficient of 0.2.