To further understanding of the mechanisms of development of resistance to penicillin in Streptococcus pneumoniae, and the role of penicillin-binding proteins (PBPs) mutations to antibiotics resistance a meta-analysis was performed. Major databases, Pubmed, Current Contents, Biosis previews, Web of Science, were searched for studies that published within 1997 through to 2007, and reported the penicillin MIC and the alteration of PBP 1a, 2b and 2x (genes or proteins) of clinical S. pneumoniae isolates. Papers were reviewed by 2 persons and used standard criteria to enroll them. Meta-analysis was performed using a random-effects model. Overall, 20 studies were included in the meta-analysis. For the included 1771 clinical S. pneumoniae isolates, the susceptibility to penicillin decreased in inverse proportion to the presence of mutated pbp genes. The mutations of the conserved amino acid motifs STMK and SRNVP of PBP 1A, STMK and LKSG of PBP2X, and SSNT of PBP2B are critical for the penicillin resistance. Those motifs can be used as markers for the penicillin susceptibility of S. pneumoniae. These results are useful in helping define the mechanism of penicillin resistance in S. pneumoniae.