研究目的
Investigating the ability of MALDI-TOF MS and VITEK 2 to analyze intraocular samples from in vitro models of endophthalmitis for rapid identification and establishment of the antimicrobial susceptibility profile of the organism involved.
研究成果
Directly applying endophthalmitis vitreous samples onto MALDI-TOF MS and VITEK 2 presents a promising new technique for the rapid identification of pathogens in the setting of endophthalmitis. Further studies are needed to further validate potential clinical applications.
研究不足
The scope of pathogen identification is limited by the breadth of organisms established in the database of the specific biotyper software that is employed. For clinical applicability, there must be an adequate quantity of bacteria present in intraocular samples obtained from patients.
1：Experimental Design and Method Selection:
Vitreous humor aspirated from freshly enucleated porcine eyes was inoculated with different inocula of Staphylococcus aureus (S. aureus) and incubated at 37?C. Vitreous endophthalmitis samples were centrifuged and pellets were directly analyzed with MALDI-TOF MS and VITEK 2 without prior culture.
2：Sample Selection and Data Sources:
Vitreous humor was aspirated with an 18-gauge needle fitted onto a 10ml syringe using sterile technique.
3：List of Experimental Equipment and Materials:
MALDI-TOF MS (Vitek MS, Version
4：0, bioMérieux), VITEK 2 automated antimicrobial susceptibility test system (VITEK 2, Version 01, bioMérieux), 22μm PES membrane (Whatman, Clifton, NJ), tryptic soy agar (TSA). Experimental Procedures and Operational Workflow:
Vitreous humor was filtered, aliquoted, and stored at -80?C until use. Bacterial stock was prepared and mixed into vitreous humor to make serial dilution samples. Samples were incubated, centrifuged, and pellets were analyzed.
5：Data Analysis Methods:
MALDI-TOF MS achieved accurate pathogen identification with confidence values of up to 99.9%. VITEK 2 gave AST profiles that were up to 94.4% identical to the positive control.
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