Detection of Penicillin Resistance in Streptococcus pneumoniae by a Seminested PCR Strategy

2003 ◽  
pp. 65-75
Author(s):  
Mignon du Plessis ◽  
Anthony M. Smith ◽  
Keith P. Klugman
Keyword(s):  
Streptococcus Pneumoniae ◽  
Penicillin Resistance ◽  
Seminested Pcr

scholarly journals Application of pbp1A PCR in Identification of Penicillin-Resistant Streptococcus pneumoniae

1999 ◽  
Vol 37 (3) ◽  
pp. 628-632 ◽  
Author(s):  
Mignon du Plessis ◽  
Anthony M. Smith ◽  
Keith P. Klugman
Keyword(s):  
Streptococcus Pneumoniae ◽  
Clinical Isolates ◽  
Penicillin Resistance ◽  
Binding Region ◽  
Pcr Assay ◽  
Specific Primers ◽  
Predictive Values ◽  
Species Specific ◽  
Seminested Pcr

A seminested PCR assay, based on the amplification of the pneumococcal pbp1A gene, was developed for the detection of penicillin resistance in clinical isolates of Streptococcus pneumoniae. The assay was able to differentiate between intermediate (MICs = 0.25 to 0.5 μg/ml) and higher-level (MICs = ≥1 μg/ml) resistance. Two species-specific primers, 1A-1 and 1A-2, which amplified a 1,043-bp region of thepbp1A penicillin-binding region, were used for pneumococcal detection. Two resistance primers, 1A-R1 and 1A-R2, were designed to bind to altered areas of the pbp1A gene which, together with the downstream primer 1A-2, amplify DNA from isolates with penicillin MICs of ≥0.25 and ≥1 μg/ml, respectively. A total of 183 clinical isolates were tested with the pbp1A assay. For 98.3% (180 of 183) of these isolates, the PCR results obtained were in agreement with the MIC data. The positive and negative predictive values of the assay were 100 and 91%, respectively, for detecting strains for which the MICs were ≥0.25 μg/ml and were both 100% for strains for which the MICs were ≥1 μg/ml.

Quintuplex PCR to Detect Antibiotic Resistance Genes in Streptococcus pneumoniae

2016 ◽  
Vol 1 (2) ◽  
pp. 22 ◽  
Author(s):  
Navindra Kumari Palanisamy ◽  
Parasakthi Navaratnam ◽  
Shamala Devi Sekaran
Keyword(s):  
Antibiotic Resistance ◽  
Streptococcus Pneumoniae ◽  
Resistance Genes ◽  
Bacterial Pathogen ◽  
Antibiotic Resistance Genes ◽  
Pcr Amplification ◽  
Penicillin Resistance ◽  
Penicillin Binding Proteins ◽  
Efflux Mechanism ◽  
Mic Value

Introduction: Streptococcus pneumoniae is an important bacterial pathogen, causing respiratory infection. Penicillin resistance in S. pneumoniae is associated with alterations in the penicillin binding proteins, while resistance to macrolides is conferred either by the modification of the ribosomal target site or efflux mechanism. This study aimed to characterize S. pneumoniae and its antibiotic resistance genes using 2 sets of multiplex PCRs. Methods: A quintuplex and triplex PCR was used to characterize the pbp1A, ermB, gyrA, ply, and the mefE genes. Fifty-eight penicillin sensitive strains (PSSP), 36 penicillin intermediate strains (PISP) and 26 penicillin resistance strains (PRSP) were used. Results: Alteration in pbp1A was only observed in PISP and PRSP strains, while PCR amplification of the ermB or mefE was observed only in strains with reduced susceptibility to erythromycin. The assay was found to be sensitive as simulated blood cultures showed the lowest level of detection to be 10cfu. Conclusions: As predicted, the assay was able to differentiate penicillin susceptible from the non-susceptible strains based on the detection of the pbp1A gene, which correlated with the MIC value of the strains.

scholarly journals Determination of penicillin resistance in Streptococcus pneumoniae isolates from southern Brazil by PCR

2006 ◽  
Vol 10 (2) ◽  
pp. 110-115 ◽  
Author(s):  
Eduardo Walker Zettler ◽  
Rosane Machado Scheibe ◽  
Cícero A.G. Dias ◽  
Patrícia Santafé ◽  
Diógenes Santiago Santos ◽  
...  
Keyword(s):  
Streptococcus Pneumoniae ◽  
Penicillin Resistance ◽  
Southern Brazil

scholarly journals Bacteremic Pneumonia Caused by Penicillin-Resistant Pneumococci: Case Report and Review with a Canadian Perspective

1992 ◽  
Vol 3 (4) ◽  
pp. 185-188
Author(s):  
David R Burdge ◽  
Vincent C Woo ◽  
Patricia MA Ritchie
Keyword(s):  
Antibiotic Resistance ◽  
Case Report ◽  
Streptococcus Pneumoniae ◽  
Minimal Inhibitory Concentration ◽  
Inhibitory Concentration ◽  
Penicillin Resistance ◽  
Sensitivity Testing ◽  
Antimicrobial Sensitivity ◽  
Canadian Perspective ◽  
Bacteremic Pneumonia

A Canadian adult with bacteremic pneumonia caused by a relatively penicillin-resistant (minimal inhibitory concentration 0.25 μg/mL) Streptococcus pneumoniae is reported, and the published literature regarding penicillin-resistant pneumococci in Canada reviewed. Although penicillin resistance has been reported infrequently to date, this case emphasizes the need for routine antimicrobial sensitivity testing of all pneumococci isolated from normally sterile sites, and for ongoing systematic surveillance for penicillin and other antibiotic resistance in Canada.

scholarly journals Capsule Homology Does Not Increase the Frequency of Transformation of Linked Penicillin Binding Proteins PBP 1a and PBP 2x in Streptococcus pneumoniae

2005 ◽  
Vol 49 (4) ◽  
pp. 1591-1592 ◽  
Author(s):  
Krzysztof Trzciński ◽  
Adam MacNeil ◽  
Keith P. Klugman ◽  
Marc Lipsitch
Keyword(s):  
Streptococcus Pneumoniae ◽  
Binding Proteins ◽  
Penicillin Resistance ◽  
Penicillin Binding Proteins ◽  
Transformation Rates

ABSTRACT Penicillin resistance is mainly confined to a limited number of Streptococcus pneumoniae serotypes. Given linkage between the capsular biosynthesis locus and two penicillin binding proteins, we tested whether capsule homology increases transformation rates of penicillin resistance. Transformation rates in homologous donor-recipient pairs were no higher than expected, falsifying this hypothesis.

scholarly journals Meningitis in a Canadian Adult due to High Level Penicillin-Resistant, Cefotaxime-IntermediateStreptococcus pneumoniae

1996 ◽  
Vol 7 (3) ◽  
pp. 205-207 ◽  
Author(s):  
Cécile Tremblay ◽  
Anne-Marie Bourgault ◽  
Pierre St-Antoine
Keyword(s):  
Streptococcus Pneumoniae ◽  
Adult Patient ◽  
Penicillin Resistance ◽  
High Level

Invasive penicillin-resistant pneumococcal (PRSP) infections are increasing worldwide. In Canada, the incidence of penicillin resistance amongStreptococcus pneumoniaeisolates is estimated at greater than 6%. In Quebec, only one case of PRSP meningitis has been reported and involved an infant. An adult patient is described who presented with meningitis caused by high level penicillin-resistant, cefotaxime-intermediateS pneumoniae.

scholarly journals Impact of Azithromycin Administration for Trachoma Control on the Carriage of Antibiotic-Resistant Streptococcus pneumoniae

2003 ◽  
Vol 47 (9) ◽  
pp. 2765-2769 ◽  
Author(s):  
Sarah L. Batt ◽  
Bambos M. Charalambous ◽  
Anthony W. Solomon ◽  
Charles Knirsch ◽  
Patrick A. Massae ◽  
...  
Keyword(s):  
Streptococcus Pneumoniae ◽  
Macrolide Resistance ◽  
Penicillin Resistance ◽  
Resistant Isolate ◽  
Antibiotic Resistant ◽  
Northern Tanzania

ABSTRACT Community distribution of azithromycin has an important role to play in trachoma control. Previous studies have suggested that this may increase the prevalence of macrolide-resistant Streptococcus pneumoniae. S. pneumoniae was isolated from children under 7 years of age in Rombo District, northern Tanzania, before and 2 and 6 months after community-wide administration of azithromycin. Overall carriage rates were 11, 12, and 7%, respectively. Only one macrolide-resistant isolate carrying the mef gene was obtained 6 months after azithromycin administration. This contrasted with cotrimoxazole and penicillin resistance, both of which were common (cotrimoxazole resistance, 42, 43, and 47%, and penicillin resistance, 21, 17, and 16% at baseline, 2 months, and 6 months, respectively). There was a significant association between cotrimoxazole and penicillin resistance (P < 0.0001, Fisher's exact). These data suggest that in communities where macrolide resistance is rare, azithromycin distribution for trachoma control is unlikely to increase the prevalence of resistant organisms.

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