Defining the phylogenetics and resistome of the major ribotypes circulating in Australia.

infection (CDI) remains a significant public health threat globally. New interventions to treat CDI rely on an understanding of the evolution and epidemiology of circulating strains. Here we provide longitudinal genomic data on strain diversity, transmission dynamics and antimicrobial resistance (AMR) of ribotypes (RTs) 014/020 (=169), 002 (=77) and 056 (=36), the three most prominent strains causing CDI in Australia.

Evaluation of a custom Sensititre YeastOne plate for susceptibility testing of isavuconazole and other antifungals against clinically relevant yeast and mould species in three Australian diagnostic mycology laboratories.

This study aimed to validate the performance of the custom formulated Sensititre YeastOne One (SYO) microdilution plate which includes isavuconazole (AUSNMRC1) to perform susceptibility testing on clinically relevant yeast and mould species across three Australian reference laboratories. The minimum inhibitory concentration (MIC) results were compared with the IVD approved SYO YO10 microdilution plate and isavuconazole gradient strips. A total of 127 isolates were tested on both the YO10 and AUSNMRC1 plates.

Candida auris PCR for high-throughput infection control screening.

Unlabelled: Candida auris has significant implications for infection control due to its multidrug resistance and spread in healthcare settings. Current culture-based screening methods are laborious and risk muco-cutaneous colonisation of laboratory staff. We describe the adaptation of a published real-time PCR for the identification of C.

Antimicrobial resistance surveillance of Clostridioides difficile in Australia, 2015-18.

Background: Clostridioides difficile was listed as an urgent antimicrobial resistance (AMR) threat in a report by the CDC in 2019. AMR drives the evolution of C. difficile and facilitates its emergence and spread.

Laboratory-Based Surveillance of Clostridium difficile Infection in Australian Health Care and Community Settings, 2013 to 2018.

In the early 2000s, a binary toxin (CDT)-producing strain of , ribotype 027 (RT027), caused extensive outbreaks of diarrheal disease in North America and Europe. This strain has not become established in Australia, and there is a markedly different repertoire of circulating strains there compared to other regions of the world. The Antimicrobial Resistance Surveillance (CDARS) study is a nationwide longitudinal surveillance study of infection (CDI) in Australia.

Detection and identification of dermatophyte fungi in clinical samples using a commercial multiplex tandem PCR assay.

We evaluated the performance of a commercial multiplex tandem polymerase chain reaction (PCR) for detection of dermatophytes and other fungi in skin and nail specimens by (1) testing a range of fungal and bacterial reference cultures, (2) retrospectively testing a set of skin and nail specimens with known microscopy and culture results, and (3) prospectively testing skin and nail specimens in parallel to microscopy and culture. The AusDiagnostics Dermatophytes and Other Fungi assay accurately detected and identified a range of common dermatophytes to species, species complex or genus level, as well as Candida, Aspergillus and Scopulariopsis spp. It was unable to detect uncommon dermatophytes such as Nannizzia fulva (previously Microsporum fulvum), and Paraphyton cookei (previously Microsporum cookei).

Cost-effective implementation of a custom MALDI-TOF library for the identification of South Australian Nocardia isolates.

Mass spectrometry plays a significant role in the routine identification of micro-organisms and provides the ability to incorporate newly found pathogens into the database in a cost-effective fashion. This work aims to highlight the role of mass spectrometry through improved identification of Nocardia species in a diagnostic clinical microbiology laboratory. Prior to this study we constructed a custom in-house matrix-assisted laser desorption ionisation-time of flight (MALDI-TOF) library for Nocardia isolates consisting of isolates identified to the species level.

Mycobacterial inactivation protein extraction protocol for matrix-assisted laser desorption ionization time-of-flight characterization of clinical isolates.

Background: Rapid identification of mycobacteria has been made possible with matrix assisted laser desorption ionization time-of-flight (MALDI-TOF) in recent years. Working with high concentrations of mycobacteria in a PC-3 containment facility makes MALDI-TOF cumbersome and costly. Therefore removing the inactivated isolate's protein extract from the PC-3 facility is needed for efficient identification in a routine PC-2 laboratory.

Surveillance for antimicrobial resistance in Australian isolates of Clostridium difficile, 2013-14.

Objectives: The objective of this study was to determine the activity of fidaxomicin and comparator antimicrobials against Clostridium difficile isolated from patients with C. difficile infection (CDI) in Australian hospitals and in the community.

Methods: One private and one public laboratory from five states in Australia submitted a total of 474 isolates/PCR-positive stool samples during three collection periods in August-September 2013 (n = 175), February-March 2014 (n = 134) and August-September 2014 (n = 165).

First human case of fatal Halicephalobus gingivalis meningoencephalitis in Australia.

Halicephalobus gingivalis (previously Micronema deletrix) is a free-living nematode known to cause opportunistic infections, mainly in horses. Human infections are very rare, but all cases described to date involved fatal meningoencephalitis. Here we report the first case of H.

Emergence of class 1 integron-associated GES-5 and GES-5-like extended-spectrum beta-lactamases in clinical isolates of Pseudomonas aeruginosa in South Africa.

Several different Guiana extended-spectrum (GES) enzymes have been described occurring in Enterobacteriaceae and Pseudomonas aeruginosa worldwide. Polymerase chain reaction and gene sequencing analysis confirmed bla(GES) genes identified in three P. aeruginosa clinical isolates from South Africa as bla(GES-5) and bla(GES-5)-like, respectively.

Rapid detection and sequence-specific differentiation of extended-spectrum beta-lactamase GES-2 from Pseudomonas aeruginosa by use of a real-time PCR assay.

The LightCycler was compared to nested PCR for the detection of bla(GES/IBC) genes from 100 Pseudomonas aeruginosa clinical isolates. The real-time PCR assay detected a bla(GES/IBC) gene product from 83 isolates, exhibiting a sensitivity and specificity of 94.3 and 100% respectively, compared to nested PCR and DNA sequencing.

Sequence-selective recognition of extended-spectrum beta-lactamase GES-2 by a competitive, peptide nucleic acid-based multiplex PCR assay.

Extended-spectrum beta-lactamases (ESBLs) in Pseudomonas aeruginosa, such as GES-2, which compromises the efficacy of imipenem, tend to be geographically restricted. The CC-to-AA base pair substitution at positions 493 and 494 of the bla(GES-2)-coding region distinguishes this ESBL from bla(GES-1) and the bla(IBC)-type genes, making it an ideal target for the development of a novel sequence-specific, peptide nucleic acid (PNA)-based multiplex PCR detection method. By using two primer pairs in conjunction with a PNA probe, this method provided an accurate means of identification of bla(GES-2) compared to standard PCR and gene sequencing techniques when it was used to test 100 P.

Molecular detection of GES-2 extended spectrum Beta-lactamase producing Pseudomonas aeruginosa in Pretoria, South Africa.

Screening for and detection of the novel extended spectrum Beta-lactamase (ESBL), GES-2 produced by Pseudomonas aeruginosa remains a problem in the clinical microbiology laboratory. This study aimed to compare the normally used ESBL screening agent ceftazidime, with molecular detection, to demonstrate the presence of GES-2 ESBL production in clinical isolates of P. aeruginosa.

Integrons and beta-lactamases--a novel perspective on resistance.

The understanding of microbial resistance to the beta-lactam class of antibiotics in the form of beta-lactamases has come a long way since the early discoveries of narrow-spectrum penicillinases. Integron-borne beta-lactamases co-occurring with a wide array of non-beta-lactam resistance genes, particularly pose an increasing threat to the nosocomial environment, giving rise to multi-drug resistant microbes with complex resistance patterns. Selection of potent beta-lactamases through the use of non-beta-lactam agents may be possible through integron-mediated resistance.

A nosocomial outbreak of Pseudomonas aeruginosa isolates expressing the extended-spectrum beta-lactamase GES-2 in South Africa.

Eight Pseudomonas aeruginosa clinical strains that produce the clavulanic-acid-inhibited beta-lactamase GES-2 were isolated from patients of a South African hospital from March to July 2000. They were clonally related and each harboured a 150 kb conjugative plasmid carrying a class 1 integron containing a gene cassette encoding GES-2, followed by those for beta-lactamase OXA-5 and an aminoglycoside modifying AAC(3)I-like enzyme. Hence, incidences of infection, several fatal, due to bacteria displaying clavulanate-inhibited resistance to extended-spectrum cephalosporins and reduced susceptibility to imipenem in Pretoria Academic Hospital, South Africa, can be explained, at least in part, by the spread of P.