Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been used in the field of clinical microbiology since 2010. Compared with the traditional technique of biochemical identification, MALDI-TOF MS has many advantages, including convenience, speed, accuracy, and low cost. The accuracy and speed of identification using MALDI-TOF MS have been increasing with the development of sample preparation, database enrichment, and algorithm optimization. MALDI-TOF MS has shown promising results in identifying cultured colonies and rapidly detecting samples. MALDI-TOF MS has critical research applications for the rapid detection of highly virulent and drug-resistant pathogens. Here we present a scientific review that evaluates the performance of MALDI-TOF MS in identifying clinical pathogenic microorganisms. MALDI-TOF MS is a promising tool in identifying clinical microorganisms, although some aspects still require improvement.

The infection of implanted biomaterial scaffolds presents a major challenge. Existing therapeutic solutions, such as antibiotic treatment and silver nanoparticle-containing scaffolds are becoming increasingly impractical because of the growth of antibiotic resistance and the toxicity of silver nanoparticles. We present here a novel concept to overcome these limitations, an electrospun polycaprolactone (PCL) scaffold functionalised with zinc oxide nanowires (ZnO NWs). This study assessed the antibacterial capabilities and biocompatibility of PCL/ZnO scaffolds. The fabricated scaffolds were characterised by SEM and EDX, which showed that the ZnO NWs were successfully incorporated and distributed in the electrospun PCL scaffolds. The antibacterial properties were investigated by co-culturing PCL/ZnO scaffolds with Staphylococcus aureus. Bacterial colonisation was reduced to 51.3% compared to a PCL-only scaffold. The biocompatibility of the PCL/ZnO scaffolds was assessed by culturing them with HaCaT cells. The PCL scaffolds exhibited no changes in cell metabolic activity with the addition of the ZnO nanowires. The antibacterial and biocompatibility properties make PCL/ZnO a good choice for implanted scaffolds, and this work lays a foundation for ZnO NWs-infused PCL scaffolds in the potential clinical application of tissue engineering.

• This is the first case report of diagnosing T. marneffei infection in a HIV-negative patient with the assistance of mNGS in North China. • The mNGS was used to diagnose T. marneffei infection much faster than traditional fungal culture. • This was a non-HIV patient who infected with T. marneffei.

Metagenomic next-generation sequencing (mNGS) has emerged as a revolutionary tool for infectious disease diagnostics. The necessity of mNGS in real-world clinical practice for common Lower Respiratory Tract Infections (LRTI) needs further evaluation. A total of 184 bronchoalveolar lavage fluid (BALF) samples and 322 sputa associated with LRTI were fully examined. The detection performance was compared between mNGS and standard microbiology culture, using Sanger sequencing as the reference method. 52.05% (165/317) of sputa showed identical results for all three methods. Compared to Sanger sequencing, the same results obtained by mNGS were 88.20% (284/322). In 2.80% (9/322) of cases, Sanger sequencing detected more microorganisms, while mNGS detected more in 9% (29/322) of cases. For BALF, 49.41% (85/172) of cases showed identical results for all three methods. In 91.30% (168/184) of cases, identical results were produced by both mNGS and Sanger sequencing. mNGS detected more species in 7.61% (14/184) of cases, whereas in 2.80% (2/184) instances, the Sanger sequencing detected more microorganisms than mNGS. In the 184 BALF samples, 66 samples were identified as having co-infections by mNGS, Sanger sequencing identified 64 samples, and cultures identified 22 samples. Our study demonstrates that mNGS offers a significant advantage over conventional culture methods in detecting co-infections. For common bacterial pathogens, conventional culture methods are sufficient for detection. However, mNGS provides comprehensive pathogen detection and is particularly useful for identifying rare and difficult-to-culture pathogens.

Invasive candidiasis (IC) is a severe infection primarily affecting immunocompromised patients and is associated with high mortality and substantial hospital costs. Accurate regional data on epidemiology and antifungal resistance of IC are crucial for effective clinical management. The China Hospital Invasive Fungal Surveillance Net (CHIF-NET) study is a laboratory-based multicenter study initiated in August 2009. This report presented updated data from August 2017 to December 2021, involving 76 hospitals across 28 provincial regions in China, and included a 12-year longitudinal analysis. Isolates from patients with IC were assigned to species level by matrix-assisted laser desorption ionization-time of flight mass spectrometry and internal transcribed spacer rDNA sequencing. Susceptibility testing was performed for nine antifungal agents using the broth microdilution method. A total of 11,679 Candida isolates were collected. Candida albicans was the most common Candida species (46.0%), followed by Candida tropicalis (17.6%), Candida parapsilosis sensu stricto (15.6%), and Candida glabrata sensu stricto (11.2%). Antifungal resistance rates showed notable variations among different Candida species, with the highest azole resistance observed in C. tropicalis isolates causing candidemia (42.5% to fluconazole and 38.9% to voriconazole). Cross- and multi-drug resistance to azoles and echinocandins was observed in all predominant Candida species. The species distribution of major Candida pathogens causing IC remained stable in China, while antifungal resistance increased, especially among non- albicans Candida species. Enhanced surveillance, accurate species identification, and strengthened antifungal stewardship are needed to address the growing challenge of antifungal resistance.