To systematically assess the efficacy and complications of transrectal (TR) versus transperineal (TP) prostate biopsy in the detection of prostate cancer (PCa). A meta-analysis was performed by searching the databases Pubmed, Embase and Web of science for the relevant available studies until September 1st, 2016, and thirteen studies met the inclusion criteria. The pooled odds ratios with 95% confidence intervals were calculated to evaluate the differences of TR and TP groups in PCa detection rate. Then, trial sequential analysis was performed to reduce the risk of type I error and estimated whether the evidence of the results was reliable. Overall, this meta-analysis included a total of 4280 patients, who had been accrued between April 2000 and Aug 2014 and randomly divided into TR group and TP group. Prostate biopsies included sextant, extensive and saturation biopsy procedures. Patients who received TP prostate biopsy had no significant improvement in PCa detection rate, comparing TR group. Moreover, when comparing TR and TP studies, no significant difference was found in abnormal DRE findings, serum PSA level measurement, Gleason score, prostate volume. Besides, this meta-analysis showed no obvious differences between these two groups in terms of relevant complications. Therefore, this meta-analysis revealed that no significant differences were found in PCa detection rate between TP and TR approaches for prostate biopsy. However, with regard to pain relief and additional anesthesia, TR prostate needle biopsy was relatively preferable, compared to TP prostate biopsy.