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Primary CD4 gating in lysed whole blood is evaluated for absolute CD4 counts in fresh and aged blood using an affordable compact volumetric commercial flow cytometer. It is found that primary CD4 gating in lysed whole blood using the CyFlow SL Green is an affordable and precise method for CD4 counting.

To assess the diagnostic and operational performance of the InBiOS AMD rapid diagnostic test (RDT) (Seattle, USA) for the detection of B. pseudomallei in grown blood culture broth. The InBiOS RDT is a lateral flow immunoassay in a strip format detecting B. pseudomallei capsular polysaccharide in culture fluids, marketed for research only. Broth of blood culture bottles (BacT/Alert, bioMérieux, Marcy L’Etoile, France) sampled in adult patients at the Sihanouk Hospital Center of HOPE, Phnom Penh, Cambodia, during 2010-2017 and stored at − 80 °C was tested. They included samples grown with B. pseudomallei (n = 114), samples with no growth (n = 12), and samples with growth of other pathogens (n = 139, among which Burkholderia cepacia (n = 5)). Diagnostic sensitivity and specificity were 96.5% [95% confidence interval (CI): 91.3-98.6%] and 100% [CI: 97.5-100%] respectively. Background clearance and line intensities were good and very good. The RDT’s test strip, not housed in a cassette, caused difficulties in manipulation and biosafety. The centrifugation step prescribed by the procedure challenged biosafety, but processing of 19 B. pseudomallei samples without centrifugation showed similar results for line intensity and background clearance, compared to centrifugation. The InBiOS RDT showed excellent accuracy for detection of B. pseudomallei in grown blood culture broth. Provided operational adaptations such as cassette housing, it has the potential to reduce time to diagnosis of melioidosis. [ABSTRACT FROM AUTHOR]

Laboratory procedures for blood cultures in a hospital in Phnom Penh were adapted to optimize detection of Burkholderia pseudomallei, an important pathogen in this setting. The effects of these changes are analyzed in this study. Blood cultures consisted of two BacT/ALERT bottles (bioMérieux, Marcy-l'Etoile, France). Growth was detected visually by daily inspection of the bottles. In 2016, the aerobic-anaerobic pair (FA/FN FAN) was substituted by an aerobic pair of BacT/ALERT FA Plus bottles. Blind subculture (BS) (subculture in the absence of visual growth) was advanced from day 3 to day 2 of incubation in July 2016. In July 2018, it was further advanced to day 1 of incubation. From July 2016 to October 2019, 9,760 blood cultures were sampled. The proportion of cultures showing pathogen growth decreased from 9.6% to 6.8% after the implementation of the laboratory changes (P < 0.001). Advancing the BS from day 3 to day 2 led to an increased proportion of pathogens detected by day 3 (92.8% versus 82.3%; P < 0.001); for B. pseudomallei, this increase was even more remarkable (92.0% versus 18.2%). Blind subculture on day 1 similarly increased the proportion of pathogens detected by day 2 (82.9% versus 69.0% overall, 66.7% versus 10.0% for B. pseudomallei; both P < 0.001). However, after implementation of day 1 subculture, a decrease in recovery of B. pseudomallei was observed (12.4% of all pathogens versus 4.3%; P < 0.001). In conclusion, earlier subculture significantly shortens time to detection and time to actionable results. Some organisms may be missed by performing an early subculture, especially those that grow more slowly.