Variability of the IFN-γ ELISpot assay in the context of proficiency testing and bridging studies.

Assays that assess cellular mediated immune responses performed under Good Clinical Laboratory Practice (GCLP) guidelines are required to provide specific and reproducible results. Defined validation procedures are required to establish the Standard Operating Procedure (SOP), include pass and fail criteria, as well as implement positivity criteria. However, little to no guidance is provided on how to perform longitudinal assessment of the key reagents utilized in the assay. Through the External Quality Assurance Program Oversight Laboratory (EQAPOL), an Interferon-gamma (IFN-γ) Enzyme-linked immunosorbent spot (ELISpot) assay proficiency testing program is administered. A limit of acceptable within site variability was estimated after six rounds of proficiency testing (PT). Previously, a PT send-out specific within site variability limit was calculated based on the dispersion (variance/mean) of the nine replicate wells of data. Now an overall 'dispersion limit' for the ELISpot PT program within site variability has been calculated as a dispersion of 3.3. The utility of this metric was assessed using a control sample to calculate the within (precision) and between (accuracy) experiment variability to determine if the dispersion limit could be applied to bridging studies (studies that assess lot-to-lot variations of key reagents) for comparing the accuracy of results with new lots to results with old lots. Finally, simulations were conducted to explore how this dispersion limit could provide guidance in the number of replicate wells needed for within and between experiment variability and the appropriate donor reactivity (number of antigen-specific cells) to be used for the evaluation of new reagents. Our bridging study simulations indicate using a minimum of six replicate wells of a control donor sample with reactivity of at least 150 spot forming cells per well is optimal. To determine significant lot-to-lot variations use the 3.3 dispersion limit for between and within experiment variability.

A pilot study of orthopaedic resident self-assessment using a milestones' survey just prior to milestones implementation.

OBJECTIVE: To pilot test if Orthopaedic Surgery residents could self-assess their performance using newly created milestones, as defined by the Accreditation Council on Graduate Medical Education. METHODS: In June 2012, an email was sent to Program Directors and administrative coordinators of the 154 accredited Orthopaedic Surgery Programs, asking them to send their residents a link to an online survey. The survey was adapted from the Orthopaedic Surgery Milestone Project. Completed surveys were aggregated in an anonymous, confidential database. SAS 9.3 was used to perform the analyses. RESULTS: Responses from 71 residents were analyzed. First and second year residents indicated through self-assessment that they had substantially achieved Level 1 and Level 2 milestones. Third year residents reported they had substantially achieved 30/41, and fourth year residents, all Level 3 milestones. Fifth year, graduating residents, reported they had substantially achieved 17 Level 4 milestones, and were extremely close on another 15. No milestone was rated at Level 5, the maximum possible. Earlier in training, Patient Care and Medical Knowledge milestones were rated lower than the milestones reflecting the other four competencies of Practice Based Learning and Improvement, Systems Based Practice, Professionalism, and Interpersonal Communication. The gap was closed by the fourth year. CONCLUSIONS: Residents were able to successfully self-assess using the 41 Orthopaedic Surgery milestones. Respondents' rate improved proficiency over time. Graduating residents report they have substantially, or close to substantially, achieved all Level 4 milestones. Milestone self-assessment may be a useful tool as one component of a program's overall performance assessment strategy.