Specific staphylococcal polymerase chain reaction can be a complementary tool for identifying causative organisms and guiding antibiotic management in orthopaedic infections

Saeed, Kordo; Ahmad, Nusreen; Pallett, A.; Guiver, Malcolm; Marsh, Peter

doi:10.1097/bco.0b013e3181f20439

Cited by 9 publications

(16 citation statements)

References 23 publications

(7 reference statements)

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“…The five-well multiplex real-time PCR assays were designed in conjunction with the Health Protection Agency Newcastle for this study (unpublished, S . aureus and MRSA and gap assays taken from existing literature [ 42 , 43 ]), and the primers and probe sequences used in these assays are shown in S1 Table .…”

Section: Methodsmentioning

confidence: 99%

Dental and Microbiological Risk Factors for Hospital-Acquired Pneumonia in Non-Ventilated Older Patients

Ewan

Sails

Walls³

et al. 2015

PLoS ONE

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Hospital acquired pneumonia (HAP) is often fatal in older patients. The mouth is the main reservoir of infection and studies have suggested that oral hygiene interventions may prevent HAP. The aim of this study was to investigate associations between HAP and preceding a) heavy dental plaque and b) oral carriage of potential respiratory pathogens in older patients with lower limb fracture to determine the target for intervention studies.MethodsWe obtained a time series of tongue/throat swabs from 90 patients with lower limb fracture, aged 65-101 in a general hospital in the North East of England between April 2009-July 2010. We used novel real-time multiplex PCR assays to detect S. aureus, MRSA, E. coli, P. aeruginosa, S. pneumoniae, H. influenza and Acinetobacter spp. We collected data on dental/denture plaque (modified Quigley-Hein index) and outcomes of clinician-diagnosed HAP.ResultsThe crude incidence of HAP was 10% (n = 90), with mortality of 80% at 90 days post discharge. 50% of cases occurred within the first 25 days. HAP was not associated with being dentate, tooth number, or heavy dental/denture plaque. HAP was associated with prior oral carriage with E. coli/S. aureus/P.aeruginosa/MRSA (p = 0.002, OR 9.48 95% CI 2.28-38.78). The incidence of HAP in those with carriage was 35% (4% without), with relative risk 6.44 (95% CI 2.04-20.34, p = 0.002). HAP was associated with increased length of stay (Fishers exact test, p=0.01), with mean 30 excess days (range -11.5-115). Target organisms were first detected within 72 hours of admission in 90% participants, but HAP was significantly associated with S. aureus/MRSA/P. aeruginosa/E. coli being detected at days 5 (OR 4.39, 95%CI1.73-11.16) or 14 (OR 6.69, 95%CI 2.40-18.60).ConclusionsPatients with lower limb fracture who were colonised orally with E. coli/ S. aureus/MRSA/P. aeruginosa after 5 days in hospital were at significantly greater risk of HAP (p = 0.002).

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Section: Methodsmentioning

confidence: 99%

Dental and Microbiological Risk Factors for Hospital-Acquired Pneumonia in Non-Ventilated Older Patients

Ewan

Sails

Walls³

et al. 2015

PLoS ONE

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“…To confirm the results obtained for pathogen target ArrayTube TM hybridizations, pathogen-specific RT-PCR assays were used. Multiplex meningitis-associated pathogen real-time PCR assays were constructed based on either previously published assays for many of the bacterial pathogens represented on the array e.g., Group B Streptococcus [79], H. influenzae [21,80], S. aureus [81], S. epidermidis [2], N. meningitidis [14,82] and S. pneumoniae [83], or in house designed PHE assays, where existing previously published assays were not available. Real-time PCR assays to the meningococcal capsular transport gene ctrA were used for N. meningitidis , as previously described [21].…”

Section: Methodsmentioning

confidence: 99%

Development and Assessment of a Diagnostic DNA Oligonucleotide Microarray for Detection and Typing of Meningitis-Associated Bacterial Species

et al. 2018

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Meningitis is commonly caused by infection with a variety of bacterial or viral pathogens. Acute bacterial meningitis (ABM) can cause severe disease, which can progress rapidly to a critical life-threatening condition. Rapid diagnosis of ABM is critical, as this is most commonly associated with severe sequelae with associated high mortality and morbidity rates compared to viral meningitis, which is less severe and self-limiting. We have designed a microarray for detection and diagnosis of ABM. This has been validated using randomly amplified DNA targets (RADT), comparing buffers with or without formamide, in glass slide format or on the Alere ArrayTubeTM (Alere Technologies GmbH) microarray platform. Pathogen-specific signals were observed using purified bacterial nucleic acids and to a lesser extent using patient cerebral spinal fluid (CSF) samples, with some technical issues observed using RADT and glass slides. Repurposing the array onto the Alere ArrayTubeTM platform and using a targeted amplification system increased specific and reduced nonspecific hybridization signals using both pathogen nucleic and patient CSF DNA targets, better revealing pathogen-specific signals although sensitivity was still reduced in the latter. This diagnostic microarray is useful as a laboratory diagnostic tool for species and strain designation for ABM, rather than for primary diagnosis.

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“…However, only three out of 10 patients had their antibiotics modified according to the PCR result, including escalation to broader spectrum antibiotics to cover for methicillin-resistant Staphylococcus aureus, based on a positive PCR. The rest of the cases were already on antibiotics clinically deemed appropriate and were not changed based on the PCR result [18].…”

Section: Impact Of Pcr On Antimicrobial Management In Pjimentioning

confidence: 99%

“…In most studies, it is clear that PCR has shown considerably higher sensitivity compared with conventional tissue culture for detecting pathogens in the diagnosis of PJI [6,11,[15][16][17][18][19][20][21][22][23][24] and most recent studies (published in the last 3 years) are summarized in (TABLE 1). One of the earlier prospective studies compared PCR and culture techniques from synovial fluid, which included 115 cases of potential PJI.…”

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confidence: 99%

The impact of PCR in the management of prosthetic joint infections

Saeed

Ahmad-Saeed

2015

Expert Review of Molecular Diagnostics

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An accurate diagnosis of prosthetic joint infection (PJI) remains a challenging clinical problem and is essential for the success of treatment regardless of the treatment option chosen by patients and surgeons. In recent years, PCR for the diagnosis of PJI has received much attention. Here, we review the impact of common PCR-based techniques on identifying causative organisms, antibiotic management and economics of PJI.

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Specific staphylococcal polymerase chain reaction can be a complementary tool for identifying causative organisms and guiding antibiotic management in orthopaedic infections

Cited by 9 publications

References 23 publications

Dental and Microbiological Risk Factors for Hospital-Acquired Pneumonia in Non-Ventilated Older Patients

Dental and Microbiological Risk Factors for Hospital-Acquired Pneumonia in Non-Ventilated Older Patients

Development and Assessment of a Diagnostic DNA Oligonucleotide Microarray for Detection and Typing of Meningitis-Associated Bacterial Species

The impact of PCR in the management of prosthetic joint infections

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