The invention provides a pearl fiber shirt fabric which has silky luster and is less in hairiness and anti-pilling. Warps and weft yarns of the pearl fiber shirt fabric are all 120 Ne two-fold yarns which are formed by compact spinning of 30% pearl fibers, 20% spun silks and 50% long stapled cottons. The invention further provides a preparation method of the pearl fiber shirt fabric. The preparation method of the pearl fiber shirt fabric includes the following steps: a, spinning; enabling part of combing long stapled cottons to be mixed with the spun silks and the pearl fibers and rolled in the process of opening picking, then drawing after cotton carding, mixing with other combing long stapled cottons and then through the processes of roving, fine spinning, spooling and two-for-one twisting; b, cheese dyeing; including the steps of cheese loosing, preprocessing, dyeing, hot washing, acid-washing, soaping, hot washing, cold washing, color fixing, softening, cylinder discharging, drying and cheese fastening, the preprocessing includes bleaching and enzyme treatment; c, weaving; including the steps of beaming, drafting, slashing and weaving; and d; post-processing, including the steps of desizing, softening, sizing and preshrinking.