Korean J Clin Pathol.  1999 Oct;19(5):516-512.

Detection of Penicillin-resistant Streptococcus pneumoniae by Polymerase Chain Reaction

• Lim SY
• Bae SI
• Lee CH
• Kim KD

Abstract

BACKGROUND
Resistance to penicillin of Streptococcus pneumoniae in clinical isolates has occurred by the development of altered penicillin-binding proteins (PBPs) that have greatly decreased affinity for antibiotics and has been encountered with increasing frequency in Korea. In this study, the identification of altered PBPs of S. pneumoniae in clinical isolates by using polymerase chain reaction (PCR) and relationship of PCR with the conventional antibiotic susceptibility test of penicillin were evaluated.
METHODS
Thirty isolates of S. pneumoniae from clinical specimens were used. Four sets of PCR primers of penicillin-sensitive and -resistant S. pneumoniae were designed to amplify (i) PBP 2BS: a 360 base pair fragment of the PBP 2B gene, (ii) PBP 2BCA: a 350 base pair fragment of the class A mutations present in PBP 2B gene, (iii) PBP 2BCB: a 295 base pair fragment of the class B mutations present in PBP 2B gene, and (iv) PBP 1AR: a 434 base pair fragment of the PBP 1A gene. In addition, a set of primers that amplify 273 base pair of the autolysin gene (ALY) was applied in combination with the above to identify S. pneumoniae. PCR results were compared with antibiotic susceptibility test (disk diffusion test and penicillin MIC). RESULT: Among 30 clinical isolates tested, 80% of isolates were penicillin resistant. The results of antibiotic susceptibility test were same as those of PCR methods. Among 24 penicillin resistant isolates detected by PCR methods, 5 isolates revealed PBP 2BCB gene, but 19 isolates revealed both of PBP 2BCB and 1AR genes. Five isolates with PBP 2BCB gene showed lower range of penicillin MIC (0.19 ~ 1.0 g/mL) than 19 isolates with PBP 2BCB and 1AR genes (0.75 ~ 4.0 g/mL).
CONCLUSION
Detection of altered PBP genes of S. pneumoniae by PCR may be performed for the study of penicillin resistance. This study indicates that more altered PBPs in 1AR genes are related with higher MICs.

Keyword

Streptococcus pneumoniae; PBP (penicillin binding protein); penicillin; Polymerase chain reaction

MeSH Terms

Anti-Bacterial Agents
Base Pairing
Diffusion
Korea
N-Acetylmuramoyl-L-alanine Amidase
Penicillin Resistance
Penicillin-Binding Proteins
Penicillins
Pneumonia
Polymerase Chain Reaction*
Streptococcus pneumoniae*
Streptococcus*
Anti-Bacterial Agents
N-Acetylmuramoyl-L-alanine Amidase
Penicillin-Binding Proteins
Penicillins