ABSTRACT
OBJECTIVE: To design and implement a continuing professional development (CPD) program for Cambodian ophthalmologists. DESIGN: Partnering (twinning) between the Royal Australian and New Zealand College of Ophthalmologists (RANZCO) and the Cambodian Ophthalmological Society (COS). PARTICIPANTS: Practicing ophthalmologists in Cambodia. METHODS: A conjoint committee comprising 4 ophthalmologists from RANZCO and 3 ophthalmologists from COS was established, supported by a RANZCO administrative team experienced in CPD administration. CPD requirements and recording were adapted from the RANZCO CPD framework. Cambodian ophthalmologists were surveyed during program implementation and after handover to COS. RESULTS: At the end of the 3-year program at handover to COS, a CPD program and online recording system was established. All 47 (100%) practicing ophthalmologists in Cambodia were registered for CPD, and 21/47 (45%) were actively participating in the COS CPD program online recording. Surveys of attitudes toward CPD demonstrated no significant change. CONCLUSIONS: Partnering was moderately effective in establishing a CPD program for Cambodian ophthalmologists. Uptake of CPD may have been limited by lack of a requirement for CPD for continuing medical licensure in Cambodia. Follow-up will be necessary to demonstrate CPD program longevity.
Subject(s)
Clinical Competence , Education, Medical, Continuing/methods , Health Resources , Ophthalmology/education , Program Development , Cambodia , HumansABSTRACT
Two multiplex PCR screening capabilities (TaqMan Array Cards and FilmArray) were evaluated for their ability to detect Bacillus anthracis, Francisella tularensis and Yersinia pestis in blood samples obtained from respective murine infection models. Blood samples were obtained from infected mice at 24 h intervals after exposure. Multiplex PCR results were compared with standard blood culture and singleplex real-time PCR. Across all three models, 71 mice were tested in total, within which a subset of 43 samples was shown to contain an infecting agent by at least one of the detection technologies. Within this subset of positive samples, for each model studied, the detection rates of each technology were compared. The B. anthracis model blood culture (14 of 15 agent-containing samples tested) and FilmArray PCR (12 of 15) were shown to have equivalent detection rates, which were significantly higher (at the 95â% confidence level) than singleplex (five of 14) or Array Card (two of 14) PCRs. The F. tularensis model blood culture (12 of 12) was shown to have a significantly higher (at 95â% confidence level) detection rate than all PCR technologies, with FilmArray (seven of 11) and singleplex (seven of 12) PCRs shown to have significantly higher (at 95â% confidence level) detection rates than the Array Card PCR (two of 11). Within the Y. pestis model, there was no significant difference in detection rates between blood culture (10 of 16), singleplex PCR (14 of 16), Array Card PCR (10 of 16) and FilmArray PCR (10 of 13).
Subject(s)
Bacillus anthracis/isolation & purification , Francisella tularensis/isolation & purification , Multiplex Polymerase Chain Reaction/methods , Real-Time Polymerase Chain Reaction/methods , Yersinia pestis/isolation & purification , Animals , Anthrax/blood , Anthrax/microbiology , Bacteremia/microbiology , Bacterial Load , DNA, Bacterial/genetics , Disease Models, Animal , Female , Mice , Mice, Inbred BALB C , Plague/blood , Plague/microbiology , Reproducibility of Results , Sensitivity and Specificity , Time Factors , Tularemia/blood , Tularemia/microbiologyABSTRACT
Eight DNA extraction products or methods (Applied Biosystems PrepFiler Forensic DNA Extraction Kit; Bio-Rad Instagene Only, Bio-Rad Instagene & Spin Column Purification; EpiCentre MasterPure DNA & RNA Kit; FujiFilm QuickGene Mini80; Idaho Technologies 1-2-3 Q-Flow Kit; MoBio UltraClean Microbial DNA Isolation Kit; Sigma Extract-N-Amp Plant and Seed Kit) were adapted to facilitate extraction of DNA under BSL3 containment conditions. DNA was extracted from 12 common interferents or sample types, spiked with spores of Bacillus atropheaus. Resulting extracts were tested by real-time PCR. No one method was the best, in terms of DNA extraction, across all sample types. Statistical analysis indicated that the PrepFiler method was the best method from six dry powders (baking, biological washing, milk, plain flour, filler and talcum) and one solid (Underarm deodorant), the UltraClean method was the best from four liquids (aftershave, cola, nutrient broth, vinegar), and the MasterPure method was the best from the swab sample type. The best overall method, in terms of DNA extraction, across all sample types evaluated was the UltraClean method.
Subject(s)
Bacillus/genetics , DNA, Bacterial/isolation & purification , Molecular Biology/methods , Reagent Kits, Diagnostic , Specimen Handling , Powders , Real-Time Polymerase Chain Reaction , Spores, Bacterial/geneticsABSTRACT
Real-time polymerase chain reaction (PCR) assays were developed to detect Francisella tularensis (Ft), the causative agent of tularaemia in humans. Two real-time PCRs (FTT0376 and FTT0523) were designed in genetic sequences identified by the Insignia genome comparison tool (http://insignia.cbcb.umd.edu/) as being unique to pathogenic subspecies of F. tularensis. Both PCRs identified all pathogenic F. tularensis subspecies but did not cross react with avirulent Francisella philomiragia or F. tularensis ssp. novicida or other environmental bacteria. Limits of detection from DNA purified from pure culture (FTT0376 approximately 80 Ft genome equivalents (GEs) per PCR; FTT0523 approximately 20 Ft GEs per PCR;) and DNA purified from spiked blood samples (4 x 10(4) to 4 x 10(3) cfu ml(-1), both assays) were determined.
