Determining the analytical specificity of PCR-based assays for the diagnosis of IA: What is Aspergillus?

A wide array of PCR tests has been developed to aid the diagnosis of invasive aspergillosis (IA), providing technical diversity but limiting standardisation and acceptance. Methodological recommendations for testing blood samples using PCR exist, based on achieving optimal assay sensitivity to help exclude IA. Conversely, when testing more invasive samples (BAL, biopsy, CSF) emphasis is placed on confirming disease, so analytical specificity is paramount. This multicenter study examined the analytical specificity of PCR methods for detecting IA by blind testing a panel of DNA extracted from a various fungal species to explore the range of Aspergillus species that could be detected, but also potential cross reactivity with other fungal species. Positivity rates were calculated and regression analysis was performed to determine any associations between technical specifications and performance. The accuracy of Aspergillus genus specific assays was 71.8%, significantly greater (P < .0001) than assays specific for individual Aspergillus species (47.2%). For genus specific assays the most often missed species were A. lentulus (25.0%), A. versicolor (24.1%), A. terreus (16.1%), A. flavus (15.2%), A. niger (13.4%), and A. fumigatus (6.2%). There was a significant positive association between accuracy and using an Aspergillus genus PCR assay targeting the rRNA genes (P = .0011). Conversely, there was a significant association between rRNA PCR targets and false positivity (P = .0032). To conclude current Aspergillus PCR assays are better suited for detecting A. fumigatus, with inferior detection of most other Aspergillus species. The use of an Aspergillus genus specific PCR assay targeting the rRNA genes is preferential.

Staphylococcus lugdunensis in children: A retrospective analysis.

Importance: Staphylococcus lugdunensis (S. lugdunensis) is a coagulase-negative staphylococcus (CoNS), found commonly as skin flora in humans. While most species of CoNS are clinically benign, S. lugdunensis can exhibit a similar virulence to that of S. aureus. However, there is scant data concerning S. lugdunensis infection in the pediatric population. Objective: To ascertain local S. lugdunensis infection rates and sensitivity patterns in the pediatric population. Methods: A retrospective analysis was undertaken of all S. lugdunensis isolates across a 6-year period from 2015 to 2020. Data were collected from electronic patient notes and laboratory records. Matrix-assisted laser desorption ionization and time of flight mass spectrometry were used to identify isolates. Results: Ninety-six isolates of S. lugdunensis were identified from 86 patients. Of these, 34 isolates were treated as an infection. Twenty-three (67.6%) were found to have skin as the primary source of infection. While the observed number was small, central nervous system (CNS) sources of S. lugdunensis infection appear to be a significant source: all three isolates cultured from cerebrospinal fluid were clinically managed as infection. All three were associated with ventriculoperitoneal (VP) shunt infection. No cases of S. lugdunensis infective endocarditis were identified. About 18.6% of S. lugdunensis isolates were resistant to flucloxacillin. Interpretation: S. lugdunensis is an uncommon but significant cause of infection in the pediatric population and appears to be a rising cause of CNS infection, particularly when associated with VP shunts. Flucloxacillin is recommended locally as the first choice of antibiotic.