Acetamiprid is an insecticide used on cotton, leafy vegetables, fruit trees, and other horticultural crops to control sucking insects. Acetamiprid has been classified as an unlikely human carcinogen and it has relatively low acute or chronic toxicity in mammals. As an insecticide, considerable part of applied acetamiprid would enter actually soil. However, it’s possible effects on the soil bacterial community have not be well known update. We examined bacterial community composition in an acetamiprid-polluted upland soil in China using PCR-DGGE method which can detect variation of microbial gene diversity in environments. Soils to which acetamiprid was applied at the normal field dose (0.5 mg kg^-1 dry soil), at higher dose (5mg kg-1, 25 mg kg^-1, 50 mg kg^-1) or not applied were put in pots, respectively, incubated at 28℃, and sampled weekly after incubation for PCR-DGGE analysis. Soil sample DNA was extracted using the SDS-high strength salt method and purified by CsCl2. The V3 region of 16S rDNA was amplified using the universal primers (BSF8/20 and BSR1541/20; F338GC and R518). Amplified DNA fragments were separated by parallel denaturing gradient gel electrophoresis (DGGE). The DGGE results indicated that there were differences in bacterial community diversity in upland soils treated with different concentrations of acetamiprid. There was no significant difference in DGGE patterns between 0.5 mg kg^-1 dry soil of acetamiprid and the control soil during the whole incubation time. This suggested that acetamiprid applied at the normal field dose would be unlikely to pose a threat to soil bacterial communities. However, changes were observed in DGGE fingerprints derived from soil after one week receiving the high concentrations of acetamiprid, such as 5, 25, and 50 mg kg^-1 dry soil. DNA in application-responsive bands from the acetamiprid treatments was recovered and amplified using the universal primers (F338 and R518). PCR products were recovered and cloned into pGEM-T Easy (Promega) and two clones were obtained. The two clones were sequenced using the automated Model 377 or 3730 DNA sequencing system (Applied Biosystems). The two cloned sequences had very high similarities (100%) to many uncultured bacteria reported previously in database of NCBI.