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Bowen A.C.,Charles Darwin University | Bowen A.C.,Royal Darwin Hospital | Lilliebridge R.A.,Charles Darwin University | Tong S.Y.C.,Charles Darwin University | And 8 more authors.
Journal of Clinical Microbiology | Year: 2012

Streptococcus pyogenes is commonly believed to be resistant to trimethoprim-sulfamethoxazole (SXT), resulting in reservations about using SXT for skin and soft tissue infections (SSTI) where S. pyogenes is involved. S. pyogenes' in vitro susceptibility to SXT depends on the medium's thymidine content. Thymidine allows S. pyogenes to bypass the sulfur-mediated inhibition of folate metabolism and, historically, has resulted in apparently reduced susceptibility of S. pyogenes to sulfur antibacterials. The low thymidine concentration in Mueller-Hinton agar (MHA) is now regulated. We explored S. pyogenes susceptibility to SXT on various media. Using two sets of 100 clinical S. pyogenes isolates, we tested for susceptibility using SXT Etests on MHA containing defibrinated horse blood and 20 mg/liter β-NAD (MHF), MHA with sheep blood (MHS), MHA alone, MHA with horse blood (MHBA), and MHA with lysed horse blood (MHLHBA). European Committee on Antibacterial Susceptibility Testing (EUCAST) breakpoints defined susceptibility (MIC, ≤1 mg/liter) and resistance (MIC, >2 mg/liter). In study 1, 99% of S. pyogenes isolates were susceptible to SXT on MHA, MHBA, and MHLHBA, with geometric mean MICs of 0.04, 0.04, and 0.05 mg/liter, respectively. In study 2, all 100 S. pyogenes isolates were susceptible to SXT on MHF, MHS, MHA, and MHLHBA with geometric mean MICs of 0.07, 0.16, 0.07, and 0.09 mg/liter, respectively. This study confirms the in vitro susceptibility of S. pyogenes to SXT, providing support for the use of SXT for SSTIs. A clinical trial using SXT for impetigo is ongoing. Copyright © 2012, American Society for Microbiology. All Rights Reserved.


Grando D.,RMIT University | Said M.M.,RMIT University | Mayall B.C.,Austin Pathology | Gurtler V.,RMIT University
Journal of Microbiological Methods | Year: 2012

The increased prevalence of hypervirulent ribotype 027 Clostridium difficile requires rapid identification of isolates in order to implement timely infection control strategies. High resolution melt (HRM) analysis of PCR products can identify strain variation amongst genera of bacteria. The intergenic (16S-23S rDNA) spacer region contains sequence regions conserved within genera and other sequence region variables between species within genera. We wished to investigate whether HRM analysis of PCR ribotyping products could identify ribotype 027 C. difficile. Ribotyping was performed on 93 clinical isolates and five control strains and band patterns were analysed using GelCompar II (Applied Maths, USA). Real-time PCR using ribotyping primers was performed and normalised melt curves were generated. The HRM data was then imported into ScreenClust software (QIAGEN) to generate principal component analysis graphs depicting clustered relationships of strains. Ribotyping produced clear PCR bands for 88/98 isolates tested. Dendrograms generated by GelCompar showed a diversity of ribotype patterns amongst these 88 isolates with 18 groups identified with 70% homology. One clinical isolate showed 100% homology with the control 027 strains. ScreenClust analysis of the same 88 HRM results showed clustering of isolates, with 027 strains identifiable as a unique cluster. HRM analysis correctly identified the control 027 stains and the clinical isolate shown to be 027. HRM combined with ScreenClust analysis of real-time PCR products of the 16S-23S rDNA spacer region successfully identified ribotype 027 strains. For infection control purposes this was achieved within 2-3. h of colony isolation. © 2012.


Holmes N.E.,Austin Center for Infection Research | Holmes N.E.,University of Melbourne | Trevillyan J.M.,Austin Center for Infection Research | Kidd S.E.,Womens and Childrens Hospital | Leong T.Y.-M.,Austin Pathology
Medical Mycology Case Reports | Year: 2013

We report a case of Scedosporium prolificans infection in a patient following surgery for squamous cell lung carcinoma. Combination therapy with voriconazole and terbinafine was commenced for intrathoracic infection and mycotic vasculitis. In spite of antifungal treatment, he developed culture-positive sternal and rib osteomyelitis four months later. Scedosporiosis is not commonly reported in patients with solid organ malignancies, and this case highlights its aggressive nature and propensity for direct local invasion. © 2013 International Society for Human and Animal Mycology.


Salem N.,Austin Pathology | Anderson J.J.,Austin Pathology
Pathology | Year: 2015

Direct culture onto four commercial chromogenic media, selective for the isolation of Group B Streptococcus (GBS), were compared with the conventional pre-enrichment Centers for Disease Control and Prevention (CDC) method for the ability to isolate GBS from 242 pregnant women's self-collected vaginal/perineal swabs. The sensitivities and specificities for direct culture on to chromogenic agar were 92% and 100% for StrepBSelect (Bio-Rad Laboratories), 96% and 100% for Brilliance GBS (Thermo-Fisher Scientific), 94% and 100% for CHROMagar StrepB (CHROMagar, Dutec Diagnostics), 86% and 100% for ChromID Strepto B (bioMerieux). CDC recommended broth pre-enrichment then culture on blood containing selective agar had a sensitivity and specificity of 90.0% and 100% respectively. The chromogenic agar tested produced comparable results to the pre-enrichment CDC method. Copyright © 2015 Royal College of Pathologists of Australasia. All rights reserved.


Gurtler V.,Austin Pathology | Mayall B.C.,Austin Pathology | Wang J.,Austin Pathology
Journal of Microbiological Methods | Year: 2012

A total of 2273 specimens submitted to the Austin Hospital Pathology Service for Neisseria gonorrhoeae screening between September 1, 2009 and May 11, 2011 were used in this study. Specimens were simultaneously screened and confirmed with a previously published real time PCR assay for the opa gene (extra primers were included to increase sensitivity) and the porA gene respectively. The opa gene screen and initial porA gene confirmation yielded an N. gonorrhoeae positivity rate of 0.88% (20/2273) and 0.49% (11/2191) for specimens and patients respectively. A 16S rDNA High Resolution Melt confirmatory PCR was developed subsequently; this reduced the N. gonorrhoeae positivity rate to 0.35% (8/2273) and 0.27% (6/2191) for specimens and patients respectively (not altered by 16S sequencing). The higher rate of secondary confirmation (16S HRM) in patients compared with samples was due to the detection of species other than N. gonorrhoeae detected by the initial screening and confirmation test. This underlines the importance of performing the secondary confirmatory test that has been developed in this study. © 2012.


PubMed | Austin Pathology
Type: Journal Article | Journal: Pathology | Year: 2015

Direct culture onto four commercial chromogenic media, selective for the isolation of Group B Streptococcus (GBS), were compared with the conventional pre-enrichment Centers for Disease Control and Prevention (CDC) method for the ability to isolate GBS from 242 pregnant womens self-collected vaginal/perineal swabs. The sensitivities and specificities for direct culture on to chromogenic agar were 92% and 100% for StrepBSelect (Bio-Rad Laboratories), 96% and 100% for Brilliance GBS (Thermo-Fisher Scientific), 94% and 100% for CHROMagar StrepB (CHROMagar, Dutec Diagnostics), 86% and 100% for ChromID Strepto B (bioMerieux). CDC recommended broth pre-enrichment then culture on blood containing selective agar had a sensitivity and specificity of 90.0% and 100% respectively. The chromogenic agar tested produced comparable results to the pre-enrichment CDC method.


A total of 2273 specimens submitted to the Austin Hospital Pathology Service for Neisseria gonorrhoeae screening between September 1, 2009 and May 11, 2011 were used in this study. Specimens were simultaneously screened and confirmed with a previously published real time PCR assay for the opa gene (extra primers were included to increase sensitivity) and the porA gene respectively. The opa gene screen and initial porA gene confirmation yielded an N. gonorrhoeae positivity rate of 0.88% (20/2273) and 0.49% (11/2191) for specimens and patients respectively. A 16S rDNA High Resolution Melt confirmatory PCR was developed subsequently; this reduced the N. gonorrhoeae positivity rate to 0.35% (8/2273) and 0.27% (6/2191) for specimens and patients respectively (not altered by 16S sequencing). The higher rate of secondary confirmation (16S HRM) in patients compared with samples was due to the detection of species other than N. gonorrhoeae detected by the initial screening and confirmation test. This underlines the importance of performing the secondary confirmatory test that has been developed in this study.


PubMed | Austin Pathology, Macquarie University, South Eastern Area Laboratory Services, Biochemistry and 2 more.
Type: Journal Article | Journal: The Clinical biochemist. Reviews | Year: 2016

Ineffective test follow-up is a major source of harm for patients around the world. Unreliable communication from medical laboratories (henceforth termed laboratories) to clinicians of results that represent critical or significant risk to patients (collectively termed high risk results) is a contributing factor to this problem. Throughout Australasia, management practices for such results vary considerably. The recommendations presented in this document are based on best practice derived from the published literature and follow consultation with a wide range of stakeholders. These recommendations were created to harmonise Australasian practices by guiding laboratories in the design and implementation of safe and effective communication procedures for managing high risk results which require timely notification.


PubMed | Austin Pathology, Quest Diagnostics, Macquarie University and Debrecen University
Type: Journal Article | Journal: EJIFCC | Year: 2016

Direct communication of significant (often life-threatening) results is a universally acknowledged role of the pathology laboratory, and an important contributor to patient safety. Amongst the findings of a recent survey of 871 laboratories from 30 countries by the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM), only 3 tests were noted to be common to 90% of alert lists, and only 48% of laboratories consulted clinicians in developing these alert lists despite ISO15189 recommendations to do so. These findings are similar to previous national surveys demonstrating significant variation worldwide in how critical risk results are managed and also in how these protocols are developed. In order to promote best practice and harmonization of critical risk results management, guidelines and recommendations have been published, most recently by Clinical and Laboratory Standards Institute (CLSI) and Australasian Association of Clinical Biochemists (AACB). These statements in particular have placed strong emphasis on patient risk and risk assessment in the management of critical risk results. This focus has resulted in recommendations to adopt new terminology, the consideration of risk assessment when compiling alert tables, consultative involvement of laboratory users in setting up protocols, and the need for outcome-based evidence to support our practices. With time it is expected that emerging evidence and technological improvements will facilitate the advancement of laboratories down this path to harmonization, best practice, and improve patient safety.

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