Development and validation of a risk assessment tool for BKPyV Replication in allogeneic stem cell transplant recipients.

BACKGROUND: BK polymavirus (BKPyV), a member of the family Polyomaviridae, is associated with increased morbidity and mortality in allogeneic stem cell transplant recipients. METHODS: In our previous retrospective study of 2477 stem cell transplant patients, BKPyV replication independently predicted chronic kidney disease and poor survival. In this study, using the same cohort, we derived and validated a risk grading system to identify patients at risk of BKPyV replication after transplantation in a user-friendly modality. We used 3 baseline variables (conditioning regimen, HLA match status, and underlying cancer diagnosis) that significantly predicted BKPyV replication in our initial study in a subdistribution hazard model with death as a competing risk. We also developed a nomogram of the hazard model as a visual aid. The AUC of the ROC of the risk-score-only model was 0.65. We further stratified the patients on the basis of risk score into low-, moderate-, and high-risk groups. RESULTS: The total risk score was significantly associated with BKPyV replication (P < .0001). At 30 days after transplantation, the low-risk (score ≤ 0) patients had a 9% chance of developing symptomatic BKPyV replication, while the high-risk (score ≥ 8) of the population had 56% of developing BKPyV replication. We validated the risk score using a separate cohort of 1478 patients. The AUC of the ROC of the risk-score-only model was 0.59. Both the total risk score and 3-level risk variable were significantly associated with BKPyV replication in this cohort (P < .0001). CONCLUSIONS: This grading system for the risk of symptomatic BKPyV replication may help in early monitoring and intervention to prevent BKPyV-associated morbidity, mortality, and kidney function decline.

Species-level assessment of the molecular basis of fluoroquinolone resistance among viridans group streptococci causing bacteraemia in cancer patients.

Viridans group streptococci (VGS) are a major cause of bacteraemia in neutropenic cancer patients, particularly those receiving fluoroquinolone prophylaxis. In this study, we sought to understand the molecular basis for fluoroquinolone resistance in VGS causing bacteraemia in cancer patients by assigning 115 VGS bloodstream isolates to specific species using multilocus sequence analysis (MLSA), by sequencing the quinolone resistance-determining regions (QRDRs) of gyrA, gyrB, parC and parE, and by testing strain susceptibility to various fluoroquinolones. Non-susceptibility to one or more fluoroquinolones was observed for 78% of isolates, however only 68.7% of patients were receiving fluoroquinolone prophylaxis. All but one of the determinative QRDR polymorphisms occurred in GyrA or ParC, yet the pattern of determinative QRDR polymorphisms was significantly associated with the fluoroquinolone prophylaxis received. By combining MLSA and QRDR data, multiple patients infected with genetically indistinguishable fluoroquinolone-resistant Streptococcus mitis or Streptococcus oralis strains were discovered. Together these data delineate the molecular mechanisms of fluoroquinolone resistance in VGS isolates causing bacteraemia and suggest possible transmission of fluoroquinolone-resistant S. mitis and S. oralis isolates among cancer patients.

Evaluation of an abbreviated protocol for cytomegalovirus pp65 antigenemia testing.

The cytomegalovirus antigenemia assay remains a useful tool for monitoring reactivation among transplant recipients. In this study, we compared protocols using direct lysis of small-volume, whole blood (WB) samples vs peripheral blood leukocyte (PBL)-enriched fractions. Of 363 evaluable samples, 51 (14.0%) were positive by one or both methods. Sensitivity, specificity, and negative and positive predictive values were similar (76%, 99%, 96%, and 95% vs 71%, 100%, 96%, and 100% for the WB and PBL methods, respectively). Stratification of qualitative results by WBC count revealed comparable detection rates by each method, although the total number of positive results from leukopenic samples was significantly lower than from nonleukopenic samples (P= .04). Correlation between quantitative results was high, yet the degree of clinical agreement was suboptimal. We conclude that the small-volume, WB lysis method yields results statistically comparable to that of a PBL fractionation method but with fewer technical steps and less complexity.

Precautions with gentian violet: skin marking made sterile, effective, and economical.

BACKGROUND: Surgical site infections have been caused by gentian violet (GV) marking solutions that were contaminated with Mycobaterium chelonae. GV solution is also used in surgery to mark surgical sites. It is commercially available as a solution that may not have been prepared under sterile conditions. OBJECTIVE: Our objective is to describe a skin marking method that is sterile, effective, and economical. METHODS: GV solution; microcentrifuge tubes; and round, wood toothpicks are used as an alternative to the standard surgical marker. GV (4 drops) is dispensed into a microcentrifuge tube. After capping, the tube is autoclaved. The toothpick is used as the writing instrument and dipped into the GV as needed for intraoperative skin marking. Unlike commercially available skin markers, skin moisture will not cause the writing implement (toothpick) to become ineffective; merely dry the skin before skin marking. RESULTS: Autoclaving the commercially available shelved GV solution ensures sterility. The cost of the GV, toothpicks, and microcentrifuge tubes is approximately $0.10 per operation. In contrast, commercially available surgical markers range in cost from $0.79 to $3.89 per pen (manufactured suggested retail price), a 8- to 39- fold difference. CONCLUSION: Infectious precautions should be taken with surgical site marking. Marking solutions should be prepared under sterile conditions in a pharmacy. Alternatively, commercially available nonsterile solutions can be autoclaved to ensure sterility.