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.

Objective: The objective of this study was to evaluate the performance of five commonly used automated antimicrobial susceptibility testing (AST) systems in China (Vitek 2, Phoenix, Microscan, TDR, and DL). Materials and methods: Two “unknown” isolates, S1 (ESBL-producing Escherichia coli) and S2 (KPC-producing Klebsiella pneumoniae), were sent to 886 hospitals in China for identification and AST. Using broth microdilution method (BMD) as gold standard, minimum inhibitory concentrations (MICs) were determined. Results: Most hospitals (392, 46.1%) used Vitek 2, followed by 16% each for Phoenix, Microscan, and DL systems, and 5.9% (50) used TDR system. MICs of 22 antimicrobials were evaluated for two study isolates plus three ATCC strains. Individual susceptibility results for three ATCC strains (n=1581) were submitted by 780 (91.2%) hospitals. For each AST system, 8.7% (6/69) to 13.0% (33/253) reported MICs outside the expected range for several drugs. For the two study isolates, TDR and DL systems performed the worst in MIC determination and susceptibility categorization of cefazolin and cefepime, while the Microscan system had difficulties in susceptibility categorization for aztreonam and ertapenem. Categorical agreements were >90% for most antimicrobials tested for both the isolates, among which, using BMD, no essential agreements were noted for ampicillin, piperacillin, cefazolin, cefuroxime, ceftriaxone, and trimethoprim/sulfamethoxazole. All AST systems except Vitek 2 showed unacceptable VMEs for cefazolin (S1 and S2) and major errors for ceftazidime, cefepime, and aztreonam (isolate S1), while Vitek 2 showed a high VME rate for cefepime (10.0%) and meropenem (6.2%) for S2. Conclusion: None of the five automated systems met the criteria for acceptable AST performance, but Vitek 2 provided a relatively accurate and conservative performance for most of the antimicrobials.

Purpose: In this study, we report results from a 5-year surveillance for noncandidal yeast species causing invasive infections from 65 hospitals in China. Materials and methods: Species identification was carried out by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) supplemented by rDNA sequencing, and fluconazole and voriconazole susceptibilities of yeasts were determined by Clinical and Laboratory Standards Institute (CLSI) disk diffusion methods. Results: Overall, 884 noncandidal isolates belonging to 38 species were collected. Cryptococcus neoformans was the most common (75.6%), which also comprised 96.5% of the isolates from cerebrospinal fluid (CSF) and 62.6% from blood, followed by Trichosporon asahii (6.9%) and Rhodotorula mucilaginosa (5.1%). Fluconazole susceptibility and resistant rates were 74.1% and 9.7% for C. neoformans and 81.0% and 5.2% for T. asahii. Voriconazole exhibited good activity in comparison to these two species (99.5% and 98.3% of the isolates, were susceptible). However, 100% of the R. mucilaginosa isolates were resistant to both azoles. Other noncandidal yeast species showed reduced susceptibility to fluconazole (53.3%) but most were susceptible to voriconazole (94.3%). Over the 5 years, a decrease in the proportion of fluconazole-susceptible isolates was observed for C. neoformans (90%–67%, P<0.001) and other noncandidal yeast species (91%–66%, P<0.001). Moreover, the prevalence of azole-resistant R. mucilaginosa increased from 1% to 7% (P<0.001). Conclusion: The shift in azole susceptibilities in mainland China calls for continued surveillance for noncandidal yeasts.

Objective: This study aimed to evaluate the in vitro and in vivo effects of different combinations of antimicrobial agents against carbapenemase-producing and non-producing Klebsiella pneumoniae from China. Methods: A checkerboard assay of meropenem (MEM), amikacin (AK), tigecycline (TGC), colistin (COL) and their combinations was carried out against 58 clinical carbapenem-resistant K. pneumoniae (CRKp) isolates, including 11 carbapenemase-non-producing K. pneumoniae isolates and 21 isolates producing KPC-2 enzyme, 11 NDM-1, 13 IMP, one VIM-1 and one OXA-48. The checkerboard assay was analyzed by the fractional inhibitory concentration index (FICI). A time–kill assay and Galleria mellonella infection model were conducted to evaluate the in vitro and in vivo effects of the four drugs alone and in combination. Results: In the checkerboard assay, TGC+AK and MEM+AK combinations showed the highest synergistic effect against KPC-2 and NDM-1 carbapenemase-producing isolates, with synergy+partial synergy (defined as FICI < 1) rates of 76.2% and 71.4% against KPC-2 producers, and 54.5% and 81.8% against NDM-1 producers. TGC+AK and MEM+COL combinations showed the highest rate of synergistic effect against IMP-producing isolates. Against carbapenemase-non-producing isolates, TGC+COL and TGC+AK combinations showed the highest rate of synergy effect (63.6% and 54.5%). MEM+AK showed a synergistic effect against one VIM-1 producer (FICI=0.31) and an additivite effect (FICI=1) against one OXA-48 producer. In the time–kill assay, COL+AK, COL+TGC, COL+MEM and AK+TGC showed good synergistic effects against the KPC-2-producing isolate D16. COL+MEM and COL+TGC combinations showed good effects against the NDM-1-producing isolate L13 and IMP-4-producing isolate L34. Against the carbapenemase-non-producing isolate Y105, MEM+TGC and COL+AK showed high synergistic effects, with log ₁₀CFU/mL decreases of 6.2 and 5.5 compared to the most active single drug. In the G. mellonella survival assay, MEM-based combinations had relatively high survival rates, especially when combined with colistin, against KPC-2 producers (90% survival rate) and with amikacin against metallo-beta-lactamase producers (95– 100% survival rate). Conclusion: Our study suggests that different antimicrobial agent combinations should be considered against CRKp infections with different resistance mechanisms.

Mycoplasma hominis, a well known cause of neonatal infection, has been reported as a pathogen in urogenital infections in adults; however, central nervous system (CNS) infections are rare. We report here the first case of M. hominis meningitis in China, post neurosurgical treatment for an intracerebral haemorrhage in a 71-year-old male. We describe a 71-year-old man who developed M. hominis meningitis after neurosurgical treatment and was successfully treated with combined azithromycin and minocycline therapy of 2 weeks duration, despite delayed treatment because the Gram stain of cerebrospinal fluid (CSF) yielded no visible organisms. The diagnosis required 16S rDNA sequencing analysis of the cultured isolate from CSF. Literature review of M. hominis CNS infections yielded 19 cases (13 instances of brain abscess, 3 of meningitis, 1 spinal cord abscess and 1 subdural empyema each). Delay in diagnosis and initial treatment failure was evident in all cases. With appropriate microbiological testing, antibiotic therapy (ranging from 5 days to 12 weeks) and often, multiple surgical interventions, almost all the patients improved immediately. Both our patient findings and the literature review, highlighted the pathogenic potential of M. hominis together with the challenges prompted by rare infectious diseases in particular for developing countries laboratories with limited diagnostic resources.

Background: Kodamaea ohmeri has been a rare fungal pathogen in the past decades but is now becoming more common in various invasive fungal diseases, with high mortality. There are limited data on the occurrence and distribution of K. ohmeri. Methods: Sixty-two K. ohmeri isolates collected from 24 hospitals in China over a 7-year period were studied. Performance of three phenotypic methods in the identification of this organism was assessed against a gold standard, 26S rDNA sequencing. Original identification results submitted by the participating local hospitals were reviewed. The Sensititre YeastOne YO10 (SYY) was evaluated in determining the in vitro antifungal susceptibilities using standard broth microdilution method (BMD) as a reference, and essential agreement (EA) was calculated. Results: Accurate species identification was achieved in 82.3% and 96.8% of the cases by Vitek 2 Compact and Vitek mass spectrometry (MS), respectively. For Bruker MS, 12.9% and 96.8% of the isolates were correctly identified to species level using the direct transfer and protein extraction methods, respectively. Only 29 (46.8%) isolates were initially correctly identified as K. ohmeri by the local hospitals. The highest misidentification rate (100%, 16/16) was observed in CHROMagar. According to BMD, the highest MIC₉₀ was seen in fluconazole (8 μg/mL), followed by 1 μg/mL for micafungin, caspofungin, 5-fluorocytosine, and amphotericin B, 0.5 μg/mL for itraconazole, 0.25 μg/mL for posaconazole and voriconazole. Significant differences in EAs for different drugs were observed, ranging from 95.2% for amphotericin B to 22.6% for itraconazole between SYY and BMD. Conclusion: Our study emphasizes the need for accurate identification of clinical K. ohmeri isolates and the importance of validating antifungal susceptibility by standard BMD.