Rapid screening for penicillin susceptibility of systemic pneumococcal isolates by restriction enzyme profiling of the pbp2B gene.
Restriction digest profiling of pneumococcal pbp2b-specific amplicons was effective for screening penicillin resistance. The pbp2b amplicon of all pneumococcal isolates for which the MICs of penicillin were < or = 0.03 microgram/ml had one of two different susceptible restriction profiles, and all 33 isolates for which MICs were 0.5 microgram/ml or greater had one of seven distinct resistant profiles. Low-concentration penicillin resistance (MICs = 0.06 microgram/ml to 0.25 microgram/ml) was associated with sensitive HaeIII profiles in some isolates; however, RsaI profiling and pbp2b sequence analysis of such isolates revealed that some isolates contained low-level resistant pbp2b alleles, while others had susceptible pbp2b alleles. This data indicates that low-level penicillin resistance is sometimes conferred by determinants other than pbp2b.
Centers for Disease Control and Prevention, Childhood and Respiratory Infections, Atlanta, Georgia 30333, USA. beb0@cdc.gov
B
B
Beall
R R
RR
Facklam
D M
DM
Jackson
H H
HH
Starling
D000483:Alleles; D019881:Aminoacyltransferases; D001426:Bacterial Proteins; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics*; D016172:DNA Fingerprinting; D015252:Deoxyribonucleases, Type II Site-Specific; D005260:Female; D006602:Hexosyltransferases; D006801:Humans; D008297:Male; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D002267:Muramoylpentapeptide Carboxypeptidase / Q000235:genetics*; D010403:Penicillin Resistance / Q000235:genetics*; D046915:Penicillin-Binding Proteins; D010406:Penicillins / Q000494:pharmacology*; D010458:Peptidyl Transferases; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA; D013296:Streptococcus pneumoniae / Q000145:classification* / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification
D001426:Bacterial Proteins; D002352:Carrier Proteins; D046915:Penicillin-Binding Proteins; D010406:Penicillins; D019881:Aminoacyltransferases; C478701:penicillin-binding protein 2b, Streptococcus; D010458:Peptidyl Transferases; D006602:Hexosyltransferases; D015252:Deoxyribonucleases, Type II Site-Specific; D002267:Muramoylpentapeptide Carboxypeptidase