Detection of Streptococcus pneumoniae in whole blood by PCR

Zhang, Yingze; Isaacman, Daniel J.; Wadowsky, Robert M.; Rydquist-White, J.; Post, J. Christopher; Ehrlich, GD

doi:10.1128/jcm.33.3.596-601.1995

Cited by 103 publications

(44 citation statements)

References 23 publications

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“…The use of PCR as a tool for diagnosing pneumococcal pneumonia has been evaluated in both adults and children, mainly through the testing of blood samples (whole blood, serum or plasma). Using positive blood culture as the diagnostic gold standard, the reported sensitivities for detecting S. pneumoniae in blood samples have ranged from 29% to 100% in adults (6–15), and from 57% to 100% in children (16–19). When the definition of pneumococcal pneumonia also includes patients with positive sputum cultures, the reported sensitivities range from 26% to 88% (7–9).…”

Section: Common Pneumonia Pathogensmentioning

confidence: 99%

Molecular genetic methods in the diagnosis of lower respiratory tract infections

Murdoch

2004

APMIS

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Molecular diagnostic techniques, such as PCR, have become useful tools for the rapid etiological diagnosis of lower respiratory tract infections. Nucleic acid amplification tests (NAATs) have been evaluated for detecting most respiratory pathogens, and commercial assays are available for some pathogens. However, standardized protocols are needed before these assays are introduced into routine diagnostic use. For pneumonia, NAATs offer advantages over conventional tests for the detection of Mycoplasma pneumoniae, Legionella spp. and Chlamydia pneumoniae. For pneumococcal pneumonia in adults, PCR adds little to existing diagnostic tests, and is unable to distinguish pneumococcal colonization from infection when testing respiratory samples. Although less sensitive than culture‐based methods, several commercial molecular diagnostic assays have been developed for tuberculosis and are useful rapid tests for selected patients. PCR can now be considered the rapid diagnostic test of choice for pertussis and some respiratory virus infections. Further work is required to better characterize the role of molecular diagnostic tests for diagnosing lower respiratory tract infections, and to develop standard assays that can be readily adopted by routine diagnostic laboratories.

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Section: Common Pneumonia Pathogensmentioning

confidence: 99%

Molecular genetic methods in the diagnosis of lower respiratory tract infections

Murdoch

2004

APMIS

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“…Therefore, it would be adequate for detection of bacteria in CSF specimens, since 85% of CSF samples with bacterial infection contained more than 10 3 CFU of bacteria/ml (8). Although detection of bacterial pathogens in serum or whole blood by PCR has been reported (5,12,16), the universal PCR developed in this study may not have sufficient sensitivity for blood specimens, because the number of organisms in the blood is usually quite low. In one study, 25% of patients with S. aureus bacteremia and more than 50% with E. coli and P. aeruginosa bacteremia had colony counts of Ͻ1 CFU/ml of blood (4).…”

Section: Discussionmentioning

confidence: 92%

Use of PCR with Universal Primers and Restriction Endonuclease Digestions for Detection and Identification of Common Bacterial Pathogens in Cerebrospinal Fluid

Lu¹,

Perng²,

Lee³

et al. 2000

J. Clin. Microbiol.

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We have designed a universal PCR capable of amplifying a portion of the 16S rRNA gene of eubacteria, including Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus pneumoniae, Enterococcus faecium, Enterococcus faecalis, Mycobacterium tuberculosis, Legionella pneumophila, Escherichia coli, Klebsiella pneumoniae, Serratia marcescens, Enterobacter cloacae, Pseudomonas aeruginosa, Acinetobacter baumannii, Proteus mirabilis, Haemophilus influenzae, and Neisseria meningitidis.The sizes of the amplified products from various bacteria were the same (996 bp), but the restriction patterns of most PCR products generated by HaeIII digestion were different. PCR products from S. aureus and S. epidermidis could not be digested by HaeIII but yielded different patterns when they were digested with MnlI. PCR products from S. pneumoniae, E. faecium, and E. faecalis yielded the same HaeIII digestion pattern but could be differentiated by AluI digestion . PCR products from E. coli, K. pneumoniae, S. marcescens, and E. cloacae also had the same HaeIII digestion pattern but had different patterns when digested with DdeI or BstBI. This universal PCR could detect as few as 10 E. coli or 250 S. aureus organisms. Compared with culture, the sensitivity of this universal PCR for detection and identification of bacteria directly from 150 cerebrospinal fluids was 92.3%. These results suggest that this universal PCR coupled with restriction enzyme analysis can be used to detect and identify bacterial pathogens in clinical specimens.

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“…Attempts to detect S. pneumoniae directly by PCR have already been made using primers of the lytA gene or pbp2b gene from blood (21,28), otorrhea (27), and sputum (9). We further included the pbp2b gene of PRSP for combinational detection to characterize penicillin resistance and evaluated a possible method for rapid identification of PRSP by PCR.…”

Section: Discussionmentioning

confidence: 99%

Combinational detection of autolysin and penicillin-binding protein 2B genes of Streptococcus pneumoniae by PCR

et al. 1996

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PCR was used to identify penicillin resistance in 1,062 clinical isolates of Streptococcus pneumoniae. Three sets of primers were designed to amplify (i) a 240-bp fragment of the penicillin-binding protein (PBP) 2B gene (pbp2b) of penicillin-susceptible S. pneumoniae (PSSP), (ii) a 215-bp fragment of the class A mutations of the pbp2b gene present in penicillin-resistant S. pneumoniae, and (iii) a 286-bp fragment of the class B mutation. In addition, a set of primers that amplify 273 bp of the autolysin (lytA) gene was applied in combination with the above to identify S. pneumoniae. Of 621 isolates for which MICs of penicillin were Ϲ0.06 g/ml, 614 (98.9%) were ascertained as having DNA fragments amplified by the PSSP primers. Of 441 isolates for which MICs of penicillin were м0.125 g/ml, a class A mutation was detected in only 8 (1.8%), a class B mutation was detected in 310 (70.3%), and neither class A nor class B mutations were found in the remaining 123 (27.9%). However, when analysis was limited to isolates for which MICs of penicillin were м1.0 g/ml, 247 isolates (89.8%) of 275 were found to possess a class B mutation. When PBPs were analyzed in 12 isolates with unclear mutations of the pbp2b gene by using [ 3 H]benzylpenicillin, low affinity to PBP 2B was observed in them all. These findings suggest that a pbp2b mutation other than class A or class B is present in these isolates. These results also indicate that it may be possible to identify PSSP and penicillin-resistant S. pneumoniae by applying PCR using a combination of primers to detect the susceptible pbp2b gene, resistant pbp2b gene mutations, and the lytA gene.

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Detection of Streptococcus pneumoniae in whole blood by PCR

Cited by 103 publications

References 23 publications

Molecular genetic methods in the diagnosis of lower respiratory tract infections

Molecular genetic methods in the diagnosis of lower respiratory tract infections

Use of PCR with Universal Primers and Restriction Endonuclease Digestions for Detection and Identification of Common Bacterial Pathogens in Cerebrospinal Fluid

Combinational detection of autolysin and penicillin-binding protein 2B genes of Streptococcus pneumoniae by PCR

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