【Objective】 The objective was to ascertain the spatial distribution pattern of alfalfa virus disease strainand identify an appropriate sampling method.【Method】 The spatial distribution pattern of alfalfa virus ‐ diseasedplantswere studied using thediffusion index,its deviation significance(F0),aggregation indices,and the Iwao method.【Result】 The results indicated that the spatial distribution pattern of alfalfa virus‐diseased plantswere aggregate distribution,with thebasic compontent of distribution beingindividual groups. Their distribution was aggregated,showing anoticeable center of disease. The theoretical sampling quantity was determined under different diseased plant rates.Through statistical analysis and comparison of survey data obtained usingsix sampling methods (parallel line,Z ‐shaped,W ‐ shaped,five ‐ point,single ‐ diagonal and double ‐ diagonal),it was found that the parallel ‐ line samplingmethod had the smallest coefficient of variation and was more suitable for investigation the spatial distribution of alfalfavirus ‐ disease plants.【Conclusion】 Alfalfa virus ‐ diseasedplants exhibited an aggregated spatial distribution,andthe parallel‐line method was identified as the appropriate sampling technique for assessing spatial distribution patterns.