Purpose Basing on semiquantitative assessment of 99mTc-MIBI uptake in parathyroids of secondary hyperparathyroidism (SHPT) patients from chronic renal failure, objective guidance could be given to improve the qualitative diagnosis accuracy of MIBI uptake.Methods MIBI uptake intensiveness was semiquantitatively calculated with software ImageJ. MIBI uptake intensiveness and clinical indices were compared in 3-level grouping method consisting of slight (group 1), medium (group 2) and high (group 3) MIBI uptake groups and 2-level grouping method consisting of insignificant and significant MIBI uptake groups.Results Patient age, renal failure course, hemodialysis vintage, glomerular filtration rate (GFR), serum parathyroid hormone (PTH) and alkaline phosphatase (AKP) were positively, but serum uric acid (UA) was negatively, significantly related to MIBI uptake intensiveness; patient age was negatively, but serum phosphorus (P) and calcium (Ca) ´ P were positively, significantly related to MIBI washout; oral administration of calcitriol and calcium would significantly reduce MIBI uptake and washout. MIBI uptake tendency might alter during specified course. In 3-level grouping method, such 7 indices as the MIBI uptake intensiveness, renal failure course, hemodialysis vintage, serum AKP, Ca, cysteine proteinase inhibitor C and PTH were comparable between group 1 and 2, but were significantly different between group 1 and 3, and between group 2 and 3. In 2-level grouping method, above 7 indices plus blood urine nitrogen (BUN)/Creatinine were all significantly different between insignificant and significant group with these indices except BUN/Creatinine being greater in significant group than in insignificant group. All above significant relations or differences were with p < 0.05.Conclusions Patient age, renal failure course, hemodialysis vintage, GFR, serum PTH, AKP, UA, phosphorus and Ca ´ P, oral administration of calcitriol and calcium, and parathyroids themselves could significantly influence MIBI uptake in parathyroids of SHPT patients. 2-level grouping method should be adopted to qualitatively diagnosis MIBI uptake.